UNITED STATES OF AMERICA

DEPARTMENT OF HEALTH AND HUMAN SERVICES

FOOD AND DRUG ADMINISTRATION

CENTER FOR DEVICES AND RADIOLOGICAL HEALTH

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MEDICAL ADVISORY COMMITTEE

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MICROBIOLOGY DEVICES PANEL

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Thursday,

October 11, 2001

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The panel was called to order at 9:45 a.m. in Salons A-C of the Gaithersburg Hilton, 620 Perry Parkway, Gaithersburg, Maryland 20850, Dr. Michael L. Wilson, Panel Chair, presiding.

PRESENT:

DR. MICHAEL L. WILSON, Chairperson

FREDDIE M. POOLE, Executive Secretary

DR. ELLEN JO BARON, Temporary Voting Member

DR. KATHLEEN G. BEAVIS, Member

DR. KAREN C. CARROLL, Consultant

DR. PATRICIA CHARACHE, Consultant

DR. ROBERT L. DANNER, Temporary Voting Member

DR. DAVID T. DURACK, Industry Representative

DR. JANINE JANOSKY, Consultant

DR. IRVING NACHAMKIN, Consultant

DR. VALERIE L. NG, Member

DR. FREDERICK C. NOLTE, Temporary Voting Member

DR. L. BARTH RELLER, Temporary Voting Member

STANLEY M.REYNOLDS, Consumer Representative

DR. NATALIE L. SANDERS, Member

DR. JOSEPH S. SOLOMKIN, Guest

C-O-N-T-E-N-T-S

PAGE

Call to Order and Opening Remarks 3

New Business: Premarket Approval Application 9

Sponsor Presentation 10

FDA Presentation 88

Open Public Hearing 123

Open Committee Discussion 124

Open Public Hearing 152

Sponsor Response 153

FDA Response 193

Vote and Recommendations 193

New Business 207

Sponsor Presentation 208

FDA Presentation 274

Open Public Hearing 288

Open Committee Discussion 289

Adjournment 309

P-R-O-C-E-E-D-I-N-G-S

(9:47 a.m.)

CHAIRMAN WILSON: Good morning. I am Dr. Mike Wilson, Chair of the Microbiology Panel meeting and I would like to welcome everybody today. I would like to emphasize to everyone today, both on the panel and in the audience, that we have a very ambitious and full agenda for the day, currently scheduled not to end until almost seven o'clock tonight.

So we would ask everyone who is participating today to please do whatever they can to help us keep on schedule. Again, I would like to welcome everyone, and to thank everyone for coming today.

At this point, I would like to turn the meeting over to Freddie Poole, the executive secretary, for her remarking remarks.

MS. POOLE: Good morning. We have a few housekeeping reminders. If anyone has cell phones or beepers, could you please turn them off and your pages, if you could put those on vibrate just as a common courtesy.

Restrooms are just around the corner to your left, and we also have to read into the record a conflict of interest statement. The following announcement addresses conflict of interest issues associated with this meeting, and is made a part of the record to preclude even the appearance of an impropriety.

To determine if any conflict existed, the Agency reviewed the submitted agenda and all financial interests reported by the committee participants. The conflict of interest statutes prohibits special government employees from participating in matters that could affect their or their employees' financial interest.

However, the Agency has determined that participation of certain members and consultants, the need for whose services outweighs the potential conflict of interest involved, is in the best interests of the government.

Waivers have been granted for Drs. Valerie Ng and Irving Nachamkin for their financial interests in firms at issue that could potentially be affected by the panels' recommendations. The waivers allow these individuals to participate fully in today's deliberations.

Copies of these waivers may be obtained from the Agency's Freedom of Information Office, Rule 12-A15, of the Parklawn Building. We would like to note for the record that the Agency took into consideration certain matters regarding other panelists.

Drs. Ellen Baron, Karen Carroll, Frederick Nolte, Barth Reller, and Natalie Sanders, reported current or past interest in firms at issue, but in matters that are not related to today's agenda. The Agency has determined, therefore, that they may participate fully in the panel's deliberations.

In the event that discussions involve any other products or firms not already on the agenda, for which an FDA participant has a financial interest, the participants should excuse he or herself from such involvement, and the exclusion will be noted for the record.

With respect to all other participants, we ask in the interest of fairness that all persons making statements or presentations disclose any current or previous financial involvement with any firm whose products them may wish to comment upon. Dr. Wilson.

CHAIRMAN WILSON: Thank you. At this point, I would like to introduce the members of the panel. I would just like to gr around and have each person introduce themselves, and give their affiliation. I would like to start with Dr. Durack, please.

DR. DURACK: Good morning. I am Dr. David Durack, and I an the Industry Representative, and I work with Becton Dickinson, and I am also associated with Duke University.

MR. REYNOLDS: Good morning. I am Stanley Reynolds, and I am the Consumer Rep, and I am the Supervisor of the Immunology and Virology Section for the Pennsylvania State Public Health Laboratory.

DR. CHARACHE: Good morning. I am Patricia Charache, a professor of Pathology Medicine and Oncology at Johns Hopkins, where my current title is Program Director, Quality Assurance and Outcomes Research.

DR. NACHAMKIN: My name is Irving Nachamkin, and I am a Professor of Pathology and Lab Medicine at the University of Pennsylvania, and Associate Director of the Clinical Microbiology Laboratory.

DR. BARON: I'm Ellen Jo Baron, and I am the Director of the Microbiology and Virology Laboratories at Stanford University Medical Center, in the Department of Pathology and Medicine, at the Stanford University Medical School.

DR. SANDERS: I am Natalie Sanders, Assistant Clinical Professor of Medicine at USC School of Medicine, and I am a General Internist for the Southern California Permanente Medical Group, also known as Kaiser.

DR. CARROLL: Good morning. I am Karen Carroll, and I am an Associate Professor of Pathology at the University of Utah School of Medicine, and I also direct the Microbiology Laboratory for ARUP Laboratories, Incorporated, Salt Lake City.

DR. NG: Good morning. I am Valerie Ng, and I am a Professor of Laboratory Medicine and Interim Chair of the Department of Laboratory Medicine, at UC-San Francisco, and I am also the Director of the Clinical Laboratories at San Francisco General Hospital.

CHAIRMAN WILSON: As I mentioned, I am Dr. Mike Wilson, and I am from the Denver Health Medical Center, where I am the Director of the Department of Pathology and Laboratory Services, and I am also on the faculty in the Department of Pathology at the University of Colorado Health Sciences Center.

DR. BEAVIS: Good morning. I am Kathleen Beavis, and I am the Director of the Microbiology and Virology Laboratories, at Cook County Hospital, in Chicago.

DR. DANNER: Bob Danner, Critical Care Medical Department, NIH.

DR. RELLER: I am Barth Reller, Division of Infectious Diseases, Director of Clinical Microbiology, Duke University Medical Center.

DR. SOLOMKIN: Joe Solomkin, Professor of Surgery, at the University of Cincinnati College of Medicine. I am the Research Director in the Division of Trauma and Critical Care.

DR. NOLTE: Frederick Nolte, Associate Professor of Pathology and Lab Medicine, at Emory University Hospital, and Director of the Clinical Microbiology and Molecular Diagnostics Lab for Emory Medical Laboratories.

DR. JANOSKY: Janine Janosky, Associate Professor, Division of Biostatistics, Department of Family Medicine and Clinical Epidemiology, at the University of Pittsburgh.

DR. GUTMAN: And I am Steve Gutman, and I am the Director of the Division of Clinical Laboratory Devices, FDA, that is sponsoring this event.

CHAIRMAN WILSON: Thank you, and welcome to all the panel members. I appreciate everybody making the trip out for this meeting. The first order of new business for today is a pre-market approval application for Sepsis, Incorporated, Endotoxin Activity Assay, which is an in vitro diagnostic device for the determination of endotoxin activity in human blood samples, intended to rule out gram negative infection.

The first order of business will be the sponsor's presentation, and I would ask all of the panel members to please hold their questions until after all of the five presentations have been completed.

Now, the first speaker this morning will be Mr. Paul Walker, who is the President and CEO of Sepsis, Incorporated. Dr. Walker.

DR. WALKER: Mr. Chairman, and Members of the Agency, and Members of the Panel, good morning. My name is Paul Walker, and I am here this morning as the President of Sepsis, Inc., and I am here to begin our presentation on our PMA on the Endotoxin Activity Assay.

During our presentation this morning, following my introduction, we will have a discussion on the unmet medical need by Phil Dellinger; a description of the EAA device or endotoxin activity assay device by Alex Romaschin.

Our pivotal clinical trial, called the MEDIC trial, will be discussed first, Methods, by Debra Foster; and then the MEDIC results by John Marshall, and then I will make some concluding remarks.

I would like to outline the chronology of our interactions with the FDA, and they began in January of 1999, with an interactive meeting to review and discuss key elements of the clinical protocol, and the intended use claim.

On April 30th of this year, we submitted our PMA in a modular format with the manufacturing module submitted in November of 2000, and the non-clinical studies submitted in March of 2001.

In June of this year, in our FDA/PMA filing letter, we were pleased that the Agency granted our request for an expedited review based on the fact that the endotoxin activity assay may provide for earlier diagnosis over existing alternatives, which is in the best interests of public health.

Now, this setting, as you all can see, is a typical setting of an intensive care unit, where a number of members of the panel and myself have worked as clinicians for many years.

Several things are relatively obvious. The first is that patients in this setting are very sick. Patients in the intensive care unit have an overall mortality rate of 30 to 40 percent, and in fact this mortality rate has not changed in the last 20 years.

Often these patients have multiple diseases going on at the same time. They tend not to be single organ or single disease patients, but rather multiples of patients, and therefore particularly complex.

The second aspect is that things happen to these patients in a relatively short period of time. Their clinical condition may change rather dramatically in hours, as opposed to over days.

And as you can see in this picture there are a number of medical devices which are evident, and these include a respirator for chronic respiration; a dialysis machine, numerous IV pumps in order to provide the drugs and the fluids that are required to manage these complex cases, and deal with a number of disease processes that are going on at the same time.

In this setting, infection plays a very important role. Infection, acute infection, may be the reason that these patients are admitted to the intensive care unit in the first place.

But, secondly, these patients are very susceptible to developing infections during their stay in the ICU, and they are susceptible for a number of different reasons.

Because of the multiple disease processes that are going on these patients are often immunosuppressed, making these more vulnerable to bacterial infection. But the second is that because of the number of treatments that are necessary for these patients, a number of the normal mechanical barriers to infection in fact are breached, and they are breached by virtue of the therapy.

And this includes the endotracheal tube, which is necessary in most of these patients, and multiple in-dwelling intravenous or intraarterial lines, and often in-dwelling arterial catheter.

When a patient's condition changes in the intensive care unit, infection is often the first diagnosis that is suspected. But in the situation where multiple disease processes are going on, in fact we actually have very little information that in any way reduces that suspicion.

Most of the changes in fact point towards suspicion, and while we understand that the actual incident of infection is relatively low, at this moment we have very little in the way of help in order to reduce that suspicion of infection.

But with respect to infection in the intensive care unit, the diagnosis is in fact difficult, and the diagnosis is difficult because the patient's condition changes rather dramatically.

These patients may go from a relatively stable clinical presentation to a particularly unstable situation in a very short period of time. This brings forward this high presumption of the possibility of infection, but the definitive diagnosis for cultures in fact takes a period of time.

So when we look at this problem from a clinical standpoint, the development of SIRS, or Systemic Inflammatory Response Syndrome, which originally was thought to represent the development of infection, has proven to be particularly non-specific and not helpful in the analysis of these patients with respect to their possibility of infection.

So we are left essentially with the necessity for microbial cultures. Microbial cultures are challenged in this situation. As I said, these patients are often complex, and they have multiple areas that are at risk.

And therefore the first challenge to get an appropriate sample of the suspicion of infection, and this sample has to contain viable bacteria in order to allow a definitive result to be available.

And because of the challenge in actually getting a good sample, there may well be contaminating or colonization associated, which results in a number of both false negative and false positives in the use of microbial cultures in these patients.

And finally by necessity a necessity requires viable bacteria to grow up in a medium in order to be identified. By necessity this requires some time. So therefore there is a time delay between this moment of suspicion when the conditions change and the availability of the results of the cultures.

Now, in order to assess this problem and challenge this problem, there has been a great deal of understanding that has developed about the complexities of infection.

And perhaps some of the more advanced understanding is the rule that not just bacteria play in the mediation of infection, but in fact the bacterial toxins. And perhaps first and foremost in this is the rule of endotoxin.

Now, endotoxin is a fairly well described -- even by some members of our panel -- mediator or player in the area of infection, and it plays a very proximal role. So it is very early in the course of the infection that endotoxin plays its important role.

Now, in this challenge we have in trying to improve the management of patients, clearly better diagnostics and better therapeutics are important. So we have approached this problem in what is the useful of endotoxins in this situation.

In our review of the role of endotoxin in the past two specific issues have come forward, the first of which is the ability to measure endotoxin in blood in patients in the intensive care unit, particularly in a timely fashion.

And the previous assay that has been used is LAL assay, or the Limulus Amoebocyte Lysate Assay, and this has proven to be accurate in non-blood containing solutions, has proven not to be accurate in blood based on the fact that it has interfered with by proteins that are present in the blood stream.

So in order to make some advancement in this area where progress in both diagnostics and therapeutics has been particularly slow, we have adopted what we believe is a relatively model strategy.

The first is to develop an assay that will accurately, reliably, and in a timely fashion provide information on the level of endotoxin in the blood stream.

But secondly and perhaps even more importantly is to understand the role of measuring endotoxin and its relationship with infection in the intensive care unit.

Now, we know that endotoxin can be in the blood stream of patients in the intensive care, both commonly and for a number of reasons. Those reasons include that the endotoxin shed from rapidly dividing bacteria, either in the blood stream or in fact more commonly in local infection to elsewhere, and particularly by virtue of the fact that there is a large reservoir of Gram-negative organisms in the large bowel.

And that under a number of different circumstances this is translocated into the blood stream of these patients. So we recognize that the presence of endotoxin in the blood stream does not add new information or useful information with respect to infection, Gram-negative infection in patients in the intensive care unit.

But because endotoxin is so uniquely associated with Gram-negative organisms that we believe that its absence is therefore an important indicator for the absence of Gram-negative infection in these patients.

So we believe we have, and we will show you this morning, developed an endotoxin activity assay which is rapid, and it is an in vitro diagnostic, and it can be used to measure endotoxin activity in the whole blood in a timely and accurately way.

But the second part is that we would like the agency, and we would like the panel to accept perhaps a different or a shift in the paradigm strategies that are normally used in diagnostics.

Normal diagnostic testing is often used to both rule in and rule out a diagnosis. But in fact when a situation is present where a patient has one disease going on, both the rule in and the rule out component of the diagnosis may both be useful and be available.

In this situation, we are looking at the endotoxin activity assay solely as a rule out test. We at this moment cannot attach significant information with respect to infection to a positive endotoxin activity level.

On the other hand, we believe that an improvement in the patient management would be an indication of the absence of Gram-Negative infection in this patient population.

So our intended use claim is that when used in conjunction with microbial cultures and other relevant diagnostic tests, our test has indicated for use in ruling out the presence of Gram-Negative infection. Thank you.

I would now like to introduce Dr. Phillip Dellinger, who is a Professor of Medicine and Director of the Critical Care Section at Rush Medical College, Cook County, and Rush Presbyterian and St. Luke's Medical Center.

Dr. Dellinger is a renowned critical care physician and past president of the Society for Critical Care Medicine. Dr. Dellinger.

DR. DELLINGER: Thank you, Dr. Walker. I don't know how renowned I am, but I am definitely the past president, or one of the past presidents of the Society of Critical Care Medicine, but I appreciate Dr. Walker's kind words.

I am here to represent the health care professional in the intensive care unit, and also as a site investigator, I have some knowledge certainly of the MEDIC trial.

I can say as an intensive care assistant practicing for 20 years in the intensive care unit that infections in the ICU are a common reason for admission. They are potentially life threatening, unfortunately, and they are often very difficult to diagnose.

I know some of the panel members have as much experience as I do in the intensive care unit, while other panel members do not. And so I wanted to just walk you through very quickly what we do in the intensive care unit when we suspect infection.

We suspect infection, and we will call that day one, and we obtain cultures, and we almost always prescribe broad spectrum antibiotics. Based on patient risk factors, hospital infection patterns, we choose a broad spectrum of antibiotics to cover typically both Gram-positive and Gram-negative organisms.

Then we support our patient, and we step back and we hope for the best. On day two the patient has either improved, worsened, or no change. That gives us confidence that we are on the right track or sometimes concerned.

But it is not until day three, typically day three, when culture results are available, realizing that some cultures may come back a little earlier positive, and we may wait longer in some cases.

But day three is sort of the key decision point in time when we decide whether we are culture negative or culture positive. We then decide whether we are going to continue the antibiotic therapy as it is, or are we going to change it, or are we going to keep it that way and reculture, or are we going to stop it and reculture.

But it is very difficult in many cases to make this type of decisions based on just cultures. Let's now go to the MEDIC trial and let me try to integrate some of that thought process from the MEDIC trial and the results with how it could potentially help us at the bedside.

In order to get in the MEDIC trial, all patients had to have a suspicion of infection to get into this trial that measured endotoxin activity assay, and therefore by definition a hundred percent of the patients in this trial had suspicion of infection.

And you will notice that 80 percent were placed on antibiotics, and that's certainly in the ball park. Most patients do get broad spectrum antibiotics.

But I wanted to point out to you the big difference when a clinical evaluation committee adjudicated the charts blinded to the endotoxin activity assay to decide these experts whether there was confirmed infection, and whether there was Gram negative infection, you can see that only 18 percent of patients were judged to have confirmed infection.

And of the minority of those with Gram-negative, only 8 percent.

So I think the striking thing from this slide is that a hundred percent of patients with suspected infection, but only 8 percent judged to have Grand negative infection.

So I believe that clinicians do not have great confidence in the ability of currently available diagnostic tests to rule out infection across the board. Next slide.

Let's look at how an endotoxin assay that was sensitive might be useful. There is now general consensus that endotoxemia occurs in the absence of invasive Gram-negative infection, and therefore, may or may not be related to Gram-negative infection for some of the reasons that Dr. Walker mentioned, such as gut hypoprofusion.

It has been associated with Gram-positive infection, and so the specificity of measuring endotoxin may be problematic, at least currently, using either present or absent. However, with the sensitive assay for endotoxin, the absence of endotoxin in the blood stream might be very helpful for making invasive Grand-negative infection unlikely.

Back to the MEDIC trial again. On day one, cultures were obtained and the EAA test was done, and so now let's integrate those into how they may potentially affect decision making.

So we go back to day one, where we are getting cultures and prescribing broad spectrum antibiotics, and here a negative EAA, although not definitive, would still be a useful piece of information at the bedside to tailor and tune how the patient was going to be further evaluated and perhaps even some aggressiveness of treatment relative to non-Gram-negative sources, but not definitive.

But then on day three, when the culture results have returned, if both the culture and the EAA from day one are negative, with the sensitive and the toxin assay, then gram negative infection would be extremely unlikely. Next.

So, in summary, the utility of negative endotoxin activity assay would be on day one to be another piece of information with all these other pieces of information that we have at the bedside to be useful to the clinician in ascertaining suspicion of Gram-negative infection.

And on day three, when combined with negative cultures for Gram-negative organisms, would make the physician feel much, much better about the absence of invasive Gram-negative infection. Thank you.

DR. WALKER: Thank you, Dr. Dellinger. Now I will introduce Dr. Alex Romaschin, who is the Scientific Director of Sepsis. Alex is also the point of care test laboratory director for the University Health Network, and is an Associate Professor of both Laboratory Medicine and Surgery at the University of Toronto. Dr. Romaschin.

DR. ROMASCHIN: I want to thank the FDA for the opportunity to make a presentation with regards to the mechanistic aspects of this assay design.

The molecule that we have chosen as the target, namely Gram-negative endotoxin, has a unique structural property, in that the Lipid A portion of this molecule, which is the business end of the toxic

part of the molecule, which has been extensively described and chemically synthesized, is highly conserved among the pathogenic Gram-negative criteria.

The antibody that we use has high specificity and sensitivity for this part of the molecule, and so this has been our target in the assay design. Now, one of the historical problems with the detection of this molecule has been that because this single epitome is conserved, double capture anybody technique, sandwich ELISA techniques and other similar types, are inappropriate to detect this molecule, the assay that I am going to describe is a homogeneous immunoassay strategy, using biological and cellular effector molecules to recognize this structure and amplify it.

It has been well-described in the scientific literature that the presence of endotoxin is common in rapidly dividing bacteria at sites of localized infection and abscesses in the gut.

And that the presence of this molecule triggers permeability changes in epithelial and endothelial barriers, resulting in a rapid translocation of this molecule into the circulation.

So our target was to produce a highly sensitive and specific assay which would allow us to reproducibly detect this molecule. Could I have the next slide, please.

This cartoon illustrates our basic approach and it makes use of two fundamental aspects of both innate and cellular immunity, and the exclusive sensitivity that these systems have detecting antibody complexes, and amplifying their response.

In particular the IgM antibody that we have chosen recognizes the endotoxin forms a multimeric complex which is then elaborated upon by

compliment factors C3b and iC3b , which act as a mechanism increasing the signal intensity by generating these postage stamps which elaborate these complexes.

That amplification step then allows these complexes to be recognized by CR-1 and CR-3 opsonin receptors on neutrophils. The engagement of those receptors results in a up regulation of the priming of the neutrophil oxidative machinery, in terms of assembly of NADPH oxidases on the surface of the membrane.

Those interactions also amplify the response and so there is a sequential dual amplification system that is built into this assay. Now, those interactions themselves do not result in respiratory burst of the neutrophil.

And to release the oxyiradical armageddon that is present in the neutrophils, one requires a secondary stimulus, and we apply particular zymosan then to trigger the neutrophils to undergo a respiratory burst to degranulate and the concerted process of NADPH oxidases activation, and the release of myeloperoxidase, resulting in hypohalous acid production, then produces chemicals which stimulate luminal to undergo a chemiluminescent response and produce light.

So the output signal of our assay is light emission, and the other thing that I wanted to mention is that hominids, particular homo sapiens, is particularly sensitive to endotoxin among the millions of species.

So that all of the aspects of this assay gear to giving a very sensitive response. And in the next side, we depict the actual mechanistic aspects of how in practicality the assay is done.

This is a three tube assay, and the first tube of the assay design is a control tube. This tube lacks the specific antibody and in this diagram that is configured here, which is a little bit difficult to see.

But the Y-axis is the light emission, and the X-axis is the time; and the assay takes place over approximately a 20 minute interval of time. The control tube, which is the lower curve, all of these curves have a sort of pseudo-first order kinetic pattern, which is explained based on the way the assay is organized.

The control tube compensates for the intrinsic neutrophil concentration, and also the variations in reactivity that you see from patient to patient.

Tube Number 3, which is the maximum tube, which defines in every patient the potential span of response that can be made by the recognitive systems, this tube contains a maximal exogenous dose of endotoxin, and the antibody of interest.

And so this tube allows you then for each patient defined what is the maximal response magnitude that can be achieved. And in the second tube which contains only the antibody of interest, that response then interpolates between the control tube or the max tube, depending upon the magnitude, or the amount of endotoxin, that is present.

So this assay design has two important intrinsic components. It compensates for variations in neutrophil reactivity that you find in such a diverse population of patients who may have anergic neutrophils, or may have neutrophils that are highly activated by cytokine cascades.

It also takes into account the variations in neutrophils concentrations which occur in these populations. The second feature is that the calculation of endotoxin activity is a normalized calculation.

And the way this measurement is made is that the sample tube is subtracted, and the light intensity over the 20 minute period of time is subtracted from the sample tube, and also from the max tube, and that ratio then is the normalized endotoxin activity response.

We have a built-in fail safe calculation that was done on every single sample, and that is if the magnitude of this response from the maximum tube and the control tube is too small, either due to a lack of compliment protein support, or due to exhausted, highly activated neutrophils which can no longer function to recognize pre-formed immune complexes, that is recognized when the signal intensity is less than 15 percent of the max and non-assay is declared.

And that occurred in our clinical trial in less than 1-1/2 percent of the samples analyzed. So we have a way of identifying reproducibly when we have a non-assay result. And then the next slide, the question of the sensitivity of the assay I think is addressed.

We and others in the published literature have studied a wide spectrum of Gram-negative pathogens, and all of these are highly sensitively reactively antibody.

There has numerous documentation of the current affinity constant of this antibody. It is in the realm of lipopolysaccharide binding protein, which is the protein in biological systems which has the highest affinity for the Lipid A portion of endotoxin.

So this antibody has exquisite sensitivity in terms of its ability to bind both to Lipid A and the most difficult target, which is smooth LPS. In the next slide, one of the striking features of this assay is that unlike LAL and other assays which are confounded by the proteins which bind lipopolysaccharide, in fact our assay is enhanced by these proteins.

We believe the reason for this is the fact that in particular lipopolysaccharide binding protein, which is a phospholipid transfer type protein, is involved in the disaggradation of the multimiceller forms of LPS that exist in the circulation.

This creates a free pool of endotoxin for which the antibody can compete. Now, because our antibody is present several orders of magnitude higher concentration than LBP, and has similar affinity by mass action, we can compete these binding proteins to carry a powerful signal.

And so in contrast to many other assays, and in fact whole blood enhances the assay sensitivity more than a thousand-fold when you present endotoxin in the blood, as opposed to presently it in physiological buffers.

And I think this is a unique aspect of this assay which is not present in other endotoxins and assays. In the next slide, in order to address the issue of assay specificity, we can determine that we can detect endotoxin with exogenous supplementation, but what about in the actual patient ICU population.

And for this we use large doses of polymyxin to overcome the antibody, and when you add these doses, which do not interfere with either neutrophil reactivity, one sees in 18 consecutive samples with endotoxin a decline from an endotoxin activity from a mean level of .65 to a level of 0.11, which is well below the threshold of our clinical trial.

And this indicates that in patients who have in vivo endotoxin that we are able to demonstrate specificity of the assay, in terms of what we are detecting. I haven't given an exhaustive list of the Gram-positive or the fungal products.

But we have in fact studied all of the pathogenic Gram-positive bacteria, and clinical isolates, their cell laws or disruptive membrane products do not react as do pathogenic fungal products in the assay.

So the assay has high inherit specificity and it has or is designed maximally to have sensitivity at the low range of endotoxin concentrations, which may be released by bacterial infections of the Gram-negative type.

And so I believe that the unit dose format of the assay, and the repetitivity with which it can be performed, allows us to generate results within a period of an hour.

Due to the sensitivity of the assay and its specificity, the absence of endotoxsemia is then a good indication for the absence of Gram-negative infection.

DR. WALKER: Thank you very much, Dr. Romaschin, for your description of our novel assay. I would now like to introduce Debra Foster, who is the Clinical Project Manager for Sepsis, and Debra is going to describe the methodology of the MEDIC trial. Debra.

MS. FOSTER: Good morning to the panelists and Members from the FDA. I suppose we will be leaving the benchside now and going back to the bedside, and I will describe the clinical investigative plan for the endotoxin activity assay.

We have simplified a rather complicated protocol title to these five letters, M-E-D-I-C, or the MEDIC trial, and the acronym stands for Multicenter Endotoxin Detection In Critical Illness.

And that essentially describes what we were trying to accomplish with our protocol. The organizational structure behind the development and the implementation of the MEDIC trial is as follows.

Sponsor data management occurred in Toronto, Ontario, Canada, and consisted of a core group of people who supported both the clinical and the laboratory aspects of the trial.

That included training and study support throughout the implementation phase of the MEDIC protocol. We also employed a scientific community which was chaired by Dr. John Marshall, who is the principal investigator for the trial, and primary author of the investigative plan.

Dr. Andy Willan is our biostatistician,

and he is here with us today, and Dr. Deborah Cook was a methods expert throughout the development, and during the implementation of our trial.

We also employed a clinical evaluation committee, and this committee was struck when we realized that our primary outcome, assessing Gram-negative infection, would need a supplemental group of clinical experts internationally renowned, since we were running an international trial, that would work at arm's length to evaluate the end point of infection.

Lastly, we employed contract research organizations to outsource some of the study tasks, including source data verification, once again keeping with the international flavor of the CRO part Sepsis employed, and as well we used electronic data management, and electronic data capture, using a system developed by Phoenix Data Systems, in Valley Force, Pennsylvania.

There are 10 centers that participated in the MEDIC trial. They represent three regions, but four distinct countries. From the United States, we had four centers, all academic institutions.

In Canada, there was four investigative sites as well; and one in Brussels, Belgium; and one in London, the U.K. The main features of the MEDIC protocol are as follows. It was an observational study design.

We were trying to capture the true reflection of what it took to diagnose infection in critically ill patients in the intensive care unit. We used a multinational-multicenter format.

I will repeat that it was in the intensive care unit setting, and I just want to make it clear that at all times the endotoxin activity results were kept blinded to all the clinical staff at each of the sites.

In keeping with our rule-out project and the rule-out claim that we were making for the use of this assay, the primary objective was as follows. To determine whether the use of a rapid assay for a Gram-negative endotoxin can reliably exclude the diagnosis of Gram-negative infection in the clinically ill patient population who have a suspicion of infection.

Therefore, the remainder of the design follows that format, and so as mentioned previously by Dr. Dellinger, the inclusion criteria for the trial were all ICU patients suspected of having infection.

Now, screening for this main inclusion criteria occurred on any day of the patient's stay in the ICU. So not only were we screening patients of entry into the ICU, but at any time during their stay if a suspicion of infection occurred, they were eligible for enrollment.

Now, a qualified suspicion of infection further in that was that, yes, a suspicion had to be present, but it had to be a kind of caliber that there was an order on the chart for one or more diagnostic tests for infection.

And mostly commonly that was culture, and we did not discriminate against the site of suspicion. Patients with suspicion of primary pneumonia, or an injury of abdominal focus, or even a primary bacteremia, would all be included as eligible for admission into the trial.

But we also considered the fact that other diagnostic tests would equally allow for patients to be enrolled in this trial. For example, a CT Scan or a bronchoscopy.

Now, let me just finish that up before I move on to the next slide. The one thing that I will mention though is that based on conversations that we had during the protocol development process with the FDA was their insistence that all patients, all eligible patients, have at least one blood culture in and amongst their diagnostic culture regime.

So we agreed with that and incorporated that into our protocol. So despite the fact that the patient may not have bacteremia as their initial site of suspicion, we did have a protocol mandate for at least one set of blood cultures to be included. Next slide.

Patients could not be included int he trial if they met one of the following four exclusion criteria. They were known von Willebrand's disease; a massive blood transfusion defined more carefully as greater than three units of pack cells.

I will just further qualify this statement to say that we did put a six hour time window on that exclusion criteria to account for patients who have gone to the operating room, and perhaps have received three units of blood.

And we agreed that they could still be eligible for the trial if you waited for six hours, and they were still eligible at that point and could be included.

Patients undergoing plasmapheresis were not to be included in the trial, and if a patient had already participated in a trial of an anti-endotoxin therapy, then that was also exclusion criteria.

And I will just mention now in reviewing the screening records that all the sites kept for these criteria that the number of patients who did not get enrolled in the trial were less than 10 percent of all screened.

So we did not unduly influence the population by having a exclusion rate. Once patients met the inclusion criteria, and none of the exclusion criteria, they were eligible for enrollment.

Recalling the date of enrollment, Study Day One, and on Study Day One that was the day where all the microbial cultures or other diagnostic tests were performed, keeping in mind that this was the day of suspicion.

If part of the diagnostic tests ordered by the clinician did not include a blood culture, once a patient was enrolled in the trial, we asked that a blood culture be drawn.

A sample for the endotoxin activity assay was taken on the same day, and as well we collected other demographics and clinical variables in a intra-electronic data management system, and they included age, race, and gender of the patients, hospital ICU admission and discharge dates, a severity of illness indicator, known as the APACHE II score, Acute Physiologic, Age, and Chronic Health Evaluation Number II.

A score was used and organ disfunction scores were captured for these patients as well. We followed the patients for as long as seven days, or until they were discharged from the intensive care unit.

The primary study end-point then in keeping with our theme was the absence of Gram-negative infection on study day one. Now, the methods we used to evaluate that end-point were complicated, and we used a step-wise fashion.

In trying to keep with a more subjective interpretation of culture results, we initially employed an adaptive version of the Centers for Disease Control Criteria.

They were adapted to be more pertinent to the ICU patient population. However, in the course of writing a pilot study, a six week pilot study of 70 patients, we realized that a supplemental infection evaluation that included more of a clinical component would be necessary.

Therefore, we struck this clinical evaluation committee to act as a supplemental or secondary reviewer for the primary study end-point. I want to reemphasize that they were maintained as blind to the endotoxin activity results during the time of their adjudication process, and they were kept at arm's length from the core study personnel.

There will be a little bit more information on the results of the CEC versus the CDC adjudication given by Dr. Marshall. And that will therefore conclude the methods section. Thank you.

DR. WALKER: Thank you, Debra. I would now like to introduce Dr. John Marshall, who is a Professor of Surgery at the University of Toronto, and is the Research Director for the Medical Surgical Intensive Care Unit at Toronto General Hospital.

And Dr. Marshall is the principal investigator in our trial, and is a well-known authority in the area. John.

DR. MARSHALL: Thank you very much, Dr. Walker. This slide summarizes the participating sites in the study, and as Debra commented, we had 10 different sites across four countries, representing a group of primary academic and tertiary care intensive care units, and variable rates of accrual of the different investigative sites.

Now, we enrolled a total of 529 patients, and these were patients who were consented and enrolled in the study. Of those 529 patients, 64 were excluded from the evaluation because for one reason or another there was not reliable endotoxin activity data available.

This could be because the sample was missed because of problems with the baseline or maximum stimulated values on the controls, or because of equipment failure.

So we ended up with a total of 465 patients, for whom we had reliable endotoxin activity data available. We made a decision to focus only on 408 patients, and excluded 57 of those. The primary reason for these exclusions were major protocol violations.

And virtually all of them are patients who did not have the protocol mandated baseline blood cultures. And so in discussing the results, I will be focusing primarily on the 408 evaluable patients, but I will comment briefly on the population.

This shows the overall study population of 529 patients, and you have to recognize that this is a typical ICU population, and a mean age of approximately 60 years, and a predominance of males to females.

Typically this was a 60 to 40 and we found that as well. There is a sick population reflected in a number of variables, an ICU stay that averaged 14 days prolonged hospital stay; and significantly a 28 day all-cause mortality rate of 28 percent.

Now, as I mentioned, we did exclude 121 patients from the analysis that I am going to report, and it was important to make sure that there was not a systematic difference between the patients that were included and those that were excluded.

And what we did then was a multi-varied analysis to look at the variables that might differ between those two populations. The two that in fact did differ was race. There were more caucasians in those patients who were excluded, and in APACHE II, those patients were slightly sicker.

Now, in order to be sure that this was not going to bias the results, we evaluated the impact of race and APACHE II score on the relationship between endotoxin activity assay and Gram-negative infection, and in fact the relationship was such that excluding these patients would if anything underestimate the negative predictive value of the assay.

So we are comfortable that the exclusion of these patients did not positively bias the results. If anything, it negatively biased them, and probably had no consequence. Next slide.

Now, as several people have alluded to, we developed a CEC, a clinical evaluation committee, to adjudicate our primary outcome, and this was done out of necessity because there simply is not a diagnostic gold standard for the presence of infection in critically ill patients.

We went through a long process of modifying and compiling previous criteria as put forth by the CDC, but these are primarily developed to establish diagnoses of infection in non-ICU patients, and the utility in a complex critically ill population is substantially less.

So we felt that it was important that in addition to having an objective set of criteria, which is what the CDC criteria represented, to have a clinically relevant set of criteria, and to this end we developed the clinical evaluation committee, which was composed of experienced clinicians with expertise in ICU acquired infections.

The review process then was that if there was a culture positive for the patient, the cases were reviewed by two reviewers. These were Senior Fellows or Junior Faculty, and one member of the Clinical Evaluation Committee.

Consensus here resulted in consensus on the diagnosis, and disagreement at any level led to a review by a second CEC member. If there was concordance between these two, again there was agreements.

If there was a continuing difference of opinion, there was a full discussion by the entire clinical evaluation committee. In most cases, it was possible to achieve consensus at one of these two levels.

But we did have a number of cases that in fact had to be debated at some length, probably in the vicinity of about 20 or 25 cases, that required a full discussion by the CDC. That is both Gram-positive and Gram-negative infections.

Now, these are the data then focusing on patients with Gram-negative cultures. So any Gram-negative organism isolated from cultures occurred in 73 patients, or 18 percent of the study population.

CDC criteria were met by 54 of those patients, or 13 percent; and the clinical evaluation committee adjudicated that 33 of those patients in fact had clinically relevant Gram-negative infections, or roughly 60 percent of the numbers that were adjudicated by CDC criteria.

This slide shows the sites of infections, and you will appreciate that there is a preponderance of infections involving the lung by CDC criteria, and the second most common site is flood, and then there is a mixture of wound, deep site infections, urinary tract infections, and skin and soft tissue infections.

We evaluated them, the performance of the assay, using the criteria of negative predictor value because our objective here was to rule out infection in patients who had a negative endotoxin activity assay.

By CDC criteria, the negative assay had a 91 percent negative predictive value, with confidence in the range of 84 to 96 percent; and by CEC criteria, which was somewhat more restrictive, it was 94 percent.

Specificity was approximately a third, 33 percent or 32 percent here, and the sensitivity was approximately 80 percent. This is in those patients who had blood cultures and were done according to protocol.

In the population that had endotoxin activity data, but may have had protocol violations, we in fact saw similar data for negative predictive values, and again 91 percent by CDC criteria, and 94 percent by CEC criteria, and comparable specificity caused for both sensitivity.

And of course a low positive predictive value because of the sensitivity of the assay, and its lack of specificity. So we would interpret the data as follows.

That using clinical criteria -- in other words, the expect judgment on a group of senior trainees and experienced clinicians -- that a negative endotoxin activity assay, or in other words, a level of less than .4, is consistent with the conclusion that Gram-negative infection is not present in 120 of the 128 patients in whom that suspicion arose.

In other words, 94 percent of those patients. If we use objective criteria not defined by clinical expertise, namely the CDC criteria, again a negative result is consistent with the absence of disease in 117 of 128 patients, or 91 percent of patients.

Now, we did of course miss some patients, and this slide summarizes in a very abbreviated form those who were missed by CDC criteria. There were a total of eight, and in fact one of them was the same patient missed on two separate occasions; a woman who had been in the ICU for over a month when she was first studied.

And I think it is important to note that 7 of those 8 patients survived the ICU stay, and so primarily the patient population had an increased risk. So of them were clearly missed.

They had infections that when you went back and looked at them that one would conclude that this was a Gram-negative infection. One of them was a patient who was mis-classified, and some of them had infections that when we looked at it there was a question about it.

And in fact of these eight patients, two of them were in fact not treated with antibiotics for Gram-negative organisms, and improved. I think what it simply emphasizes is the inherent complexity and uncertainty of establishing a definitive diagnosis of infection in a complex population of critically ill patients. Next slide.

So, just to summarize then. The way that we would see this endotoxin activity assay being applied in the clinical context where a clinician is confronted with a patient, and for a number of reasons, he or she has concern that they may have an infectious process going on.

We would obtain cultures and prescribe antibiotics as indicated by clinical circumstances, and perform an endotoxin activity assay. Just as we use a battery of tests to establish a diagnosis, and not only culture and x-ray results, and white counts, and temperatures, we have a large number of variables that can increase our sense of anxiety that an infection might be present.

And indeed a positive endotoxin activity assay would in no way alleviate that anxiety. On the other hand, it is difficult in the ICU setting to conclude the absence of infection with, for example, a negative chest x-ray, which typically almost never occurs, with a normal white cell count when we are concerned about both increases and decreases.

So in fact what we could decide is if the endotoxin activity level was negative on the day we took the culture, then we have a 94 percent likelihood that the patient does not have a Gram-negative infection.

And that it can incorporate that data into the clinical decision making process, and that may be something as simple as deciding this is more likely Gram-positive, and perhaps we should focus on removing a line.

It may be on the potential that the fever and white count actually reflect a drug reaction rather than an infection. It may be that the patient has an occult DVT and pulmonary embolus.

So in fact a negative assay may shift the focus to other potential causes of an inflammatory state in critically-ill patients. Over the next three days, at this point we have presumptive evidence, and over the next two days, we can use this as adjunctive support.

And if we have negative cultures and a negative endotoxin assay -- and we will have a negative endotoxin assay in approximately 30 percent of the patients -- I think we can confidently conclude that Gram-negative infection is highly unlikely to be present and respond appropriately.

And with that, I would like to conclude my comments and turn it back over to Dr. Walker. Thank you.

DR. WALKER: Thank you very much, John. I would now like to essentially sum up some of the presentations that have gone on today. I think we have demonstrated that there is clearly a diagnostic dilemma in the intensive care unit with respect to infection.

The infection is difficult to diagnose in intensive care unit patients, and clearly those members of the panel who are involved in this would recognize that this is a problem on a regular basis.

We are hampered in the intensive care unit because the clinical signs are clearly not specific. We are also limited and that is because of the cultures, and despite the fact that cultures still remain a reference standard, there is a time delay in the culture results being received by the clinician i order to help direct therapy.

The true sensitivity cannot be determined, and there is clearly a variable rate of contamination in the sampling of the area of suspicion. And in this milieu there is also the problem that there is a change in the clinical context of the patient between the day of the test and clearly the day of the results.

So we think that the endotoxin activity assay could add confidence to this time of diagnostic uncertainty, and this is in the setting where there is a high prevalence for the presumption of infection in these patients.

But in fact the reality is that the true incidents of infection is low, and therefore the ability to identify patients that do not have Gram-negative infection be ruled out the component of the diagnostic, and becomes an important contributor to these very challenging patients.

We believe that the endotoxin activity provides presumptive results in a rapid time frame. So if we look at the clinical utility of this assay, and take into account everything that we have presented this morning, I would make the following comments.

The first is that I remind the panel and the agency that because of the ubiquitous nature of endotoxin, and the multiple reasons that it may be in the blood stream, we cannot add a significant degree of information to the diagnosis of infection with a positive endotoxin activity assay.

And therefore we are looking only and claiming only that this assay is useful to rule out the presence of Gram-negative infection in these patients.

On the other hand, in patients where the suspicion is very high, and the diseases are complex, and we have made so little progress in both diagnostics and therapeutics that we believe that this test has a significant application for a significant number of patients in the intensive care unit.

So as a rule out test, we believe that the endotoxin activity assay provides on day one presumptive evidence for the absence of Gram-negative infection on the day of the suspicion.

And as our clinicians have suggested this may alter particularly the diagnostic differential diagnosis and the priorities in looking for as quickly as possible the management changes that will result in an improvement in these patients, and the survival of these patients in the intensive care unit.

But clearly we do not put this test up as a stand alone test. Stand alone tests are not terribly useful in the intensive care unit in patients who are so critically ill, with so many disease processes going on at the same time.

So, we believe that the endotoxin activity acts as an adjunct to your culture reports, which are usually received on the third day. So now you may be presented with a double-negative with respect to that patient's condition at the time of sampling.

Not only is that culture report negative, but with a negative endotoxin activity, we believe that that will add significant confidence to the clinician in order to rule out Gram-negative infection on day three, and therefore act accordingly.

So that on day three the corroboration for the absence of Gram-negative infection, in conjunction with a negative culture report, may have a significant change on the therapy directed at that patient.

This test is adjunctive, in that the culture report is available on day three, but a negative endotoxin activity, with a 94 percent negative predictive value, actually incorporates the clinical judgment of an expert panel of world experts in this area of critical care.

So, in fact in addition to the negative culture and a negative predictive value, 94 percent with a CEC or clinical evaluation adjudication in fact is adjunctive.

So in conclusion I would like to reiterate what our intended use claim is, which in an interactive way we have developed with the FDA. And that is that the endotoxin activity assay is a rapid in vitro diagnostic test that utilizes a specific modoclonal antibody to measure the endotoxin activity in an EDTA whole blood specimen.

When used in conjunction with microbial cultures and other relevant diagnostic tests, the test is indicated for us in ruling out the presence of Gram-negative bacterial infections.

The EAA is intended for patients admitted to the ICU at risk of, or suspected of having, an infection. Thank you. This concludes our presentation.

CHAIRMAN WILSON: Thank you, Dr. Walker. At this time, I would like to open this up for questions from members of the panel. I would like to remind the audience that only the panel can ask questions of any of the speakers. Dr. Charache.

DR. CHARACHE: I had a question about the experimental model. I am wondering whether the level of polymorphonuclear leukocytes had any impact on the study; if they had leukopenia or leukocytosis, whether that would impact upon it.

And also whether the level of albumin -- we have a lot of patients with low albumins in intensive care, and I know that if you add out the interaction with endotoxin can impact the albumin level can. And I am wondering about controls for those.

DR. WALKER: Should we respond to those questions now?

CHAIRMAN WILSON: Yes.

DR. WALKER: Now, I would like to have a discussion slide put forward, David. All right. Those are good questions, and we would like to answer those questions. Alex, would you come forward and answer those. We will just put up the discussion slides that would be appropriate for that particular question to be answered.

DR. ROMASCHIN: Alex Romaschin, from Sepsis, Incorporated. We studied a range of neutrophil concentrations from -- and I have difficulty with U.S. units, and so I apologize. But there would be a level in SI units from .5 times 10 to the 9th per liter, to 20 times 10 to the 9th per liter, which covers a portion of the neutropenic range.

Our normal range would be around 1.5 to 2 times 10 to the 9th per liter. So we were able to detect a significance signal in patients who were neutropenic down to 0.5 times 10 to the 9th per liter.

And I recognize that many febrile and neutopenics go below that range, and we have not studied below that range. But our experience has been that because of the way that the assays organize, and because there is a fail safe in terms of either a lack of compliment proteins or neutrophil response to generate a signal, that if there was not sufficient neutrophil activity to generate a signal that would be identified.

So we have established the range that covers neutropenia and neutrophilia over quite a broad range, but not at the lowest dimension. With regard to albumin, our studies with albumin indicate that because albumin is a binding protein that binds ubiquitously many molecules, and it has a three-fatty acid binding site which binds Lipid-A in a lose manner.

If you add -- we have tested normal individuals who have been supplemented to a level of 30 grams per liter above the normal range, and in those cases you get a demonstrable lowering of the EAA value, but it is small.

And in that process it is very rare to find super normal levels of albumin in ICU patients. At best, they are usually at the normal range or slightly above, and so we don't see this as a major problem from a biological detection standpoint.

DR. CHARACHE: Thank you.

CHAIRMAN WILSON: Dr. Durack.

DR. DURACK: Now, this question I believe also is for Dr. Romaschin. And you studied some interfering substances that might potentially have interfered with the test, but I didn't see any mention of antibodies as interfering substances, and I wondered if you have any information about antibodies which would quite likely be in the blood of some of these patients in practice.

And this could be a direct interference, or it could be indirect from the effect of antibodies on Gram-negatives elsewhere in the body.

DR. ROMASCHIN: I don't have the slide, but I can tell you that we tested 10 of the top antibiotics that are used in the ICU population, and there is a list of them here.

We have tested these at the recommended NCCLS levels, which is approximately 10 times higher than the upper level of a therapeutic dose. And we have tested them both in samples that had no exogenous endotoxin and that had exogenous endotoxin added in.

There was no interference and so we were aware of the fact that this is a huge risk in these patients with these antibiotics, but we have no evidence that they interfere. The only possible one is polymyxin, but I don't believe that is used anymore.

DR. WALKER: David, could we put up Slide 41, please. Alex, would you just speak to that.

DR. ROMASCHIN: This is in units that you may be more familiar with. So this would go from 500 to 20,000 endofils per microliter of blood, and essentially what this study showed is that the response curve has not shifted because of the built-in controls.

CHAIRMAN WILSON: Dr. Nachamkin.

DR. NACHAMKIN: Could you comment on the role of cortiosteroids and the suppression response in immunoacid?

DR. WALKER: Could we have Slide 39, David, please.

DR. ROMASCHIN: In the initial documents that we submitted to the FDA, we encountered some interferences from steroids. We now believe that these interferences were due to additives in the steroid preparations that we used, which acted as a scavenger.

And so we were very careful when we repeated these studies to get pure pharmacological grade suspensions of material, and you can see that does that would approximate the largest dose that clinically that is not used in a transplant patient, those doses we saw no interference.

The only effect that we saw from these high levels of steroids was that the steroids have a chemical scavenger effect and they lower the signal of the highest doses by about 10 to 15 percent.

But this is in each tube, and this is compensated for, and so this would be similar to adding a huge dose of Vitamin C. So what they do is that they attenuate the magnitude of the signal by 10 to 15 percent, but that is the equivalent in every tube, and on the end result there is no effect.

DR. WALKER: David, could you put up Slide 40 as well.

DR. ROMASCHIN: And this is one of the problems, because this assay is highly sensitive, and you have to be very careful of what additives you add, together with the target drug.

And it took us a while with all of our test solutions, because many of these are contaminated with endotoxin. A lot of bilirubin and other interferences, for instance, that are commercially available, were very difficult to find in a pure endotoxin pure form.

DR. WALKER: This is the results from the clinical trial and there were 101 patients that were receiving significant steroids, and in fact they all generated reportable EAA results.

So that while we were concerned for the reasons that Dr. Romaschin has said, in fact in the clinical trial, we did not find that as a problem, and did not find that as a reason that the assay would not be useful.

CHAIRMAN WILSON: Dr. Carroll.

DR. CARROLL: Yes. Along those same lines, do you have any data on granulocyte stimulating factors? Some of our patients at risk for sepsis are oncology patients who are getting GCSF, for example.

Did that in any way interfere with the assay?

DR. ROMASCHIN: Yes. We don't have any specific information on that that I can attest to.

CHAIRMAN WILSON: Dr. Sanders.

DR. SANDERS: Dr. Sanders. I would just like to clarify the issue regarding the immunosuppressants, because in the packet that we received there was a statement clearly that the immunosuppressant agents often resulted in a non-test.

So I just want to be sure that I am hearing that immunosuppressant agents do not interfere with --

DR. WALKER: Could I just make a comment?

CHAIRMAN WILSON: Yes.

DR. WALKER: Following our submission, we were asked a series of questions and asked to go back and look at that. So I would ask Alex to speak about that, but we do not feel that the presence of steroids or other of the immunosuppressing are in fact a contraindication.

We were concerned, and we now have both in vitro and in vivo data that suggests that is not a concern.

DR. ROMASCHIN: Yes. We went back and redid all those studies using the purest preparations of the corticosteroids that we could get at much higher doses that were in the initial submission. And none of those interfered with the studies.

DR. SANDERS: I actually have another question, and I don't know if this is the appropriate time, but it has to do with the exclusion criteria, or should I wait? All right. And the question has to do with why was von Willebrand's disease an exclusion?

DR. ROMASCHIN: This was very early on in our studies. There is in the literature and in our -- in the particular patient that we studied, we got a non-response.

And there is some information that patients with von Willebrand's disease have a compliment disorder as well. So we simply out of safety excluded those, because it was so hard to find to study that we felt that this was a reasonable exclusion.

CHAIRMAN WILSON: Next, Dr. Reller.

DR. RELLER: I have a question for Dr. Walker, Dellinger, or Foster. What would you recommend to the clinician, or as a clinician would you do differently? What action would you take based on a positive or negative test?

DR. WALKER: Well, I will answer the easy part of that, and get some help with the other parts. I think the issue is that we believe that a positive value right now adds no information with respect to the presence or absence of infection.

So we are going to make no claims on what a positive value means. With respect to a negative value, I think both Dr. Marshall and Dr. Dellinger have suggested that the EAA would help close that gap between suspicion of infection and the reality of infection.

And give presumptive information in a very early time frame. I think perhaps that may be reflected in a different focus or direction of investigations. A patient changes their status, and is potentially septic.

I guess when a patient changes their status in the intensive care unit, infection is probably what comes first, and therefore investigations in the management are essentially directed towards that.

But as we have identified, there is a time delay in knowing the answer to that question. So what we are suggesting is that on day one with that information present that the chances of having a Gram-negative infection in that patient are relatively unlikely.

And that then perhaps more focus would be placed on both, particularly diagnostic procedures, that would help elucidate what the other potential causes are.

So if a Gram-negative infection is unlikely, it then makes you investigate or suggest that you investigate perhaps more vigorously other potential causes.

I think both Dr. Marshall and Dr. Dellinger have suggested that there is a wide range of possibilities that would explain this sudden change in patient status. So that would be my comment. John, or Phil?

DR. MARSHALL: This is Dr. Marshall speaking. I think it is a very hard question. I think that an analogy might be appropriate. Suppose we had a patient who at the time that we suspect infection we have evidence of a chest x-ray infiltrate, and the temperature of 38.2 degrees.

We do a white cell count, and depending on the white cell count, our behavior may vary. Suppose the white cell count is low, and we may see that patient as maybe immunocompromised, and want to treat them with antibiotics.

If it is normal, we may say this chest x-ray infiltrate is probably simply fluid. If the white cell count is higher, our center of gravity would be shifted towards perhaps doing a diagnostic test to look for broncho alveolar lavage, or something to look for a pathogen and the like.

I think in the same way an endotoxin activity assay at day one is simply an additional piece of information that might shift the fulcrum.

So if that test is positive, I think as Dr. Walker says, we simply -- there is too much noise.

Seventy percent of the patients will be positive, and we can't draw conclusions from that any more than we can draw conclusions from a white count of 12,000.

On the other hand, if the test is negative, we may then be inclined to say that this is more likely to be a Gram-positive infection, or a non-infectious cause, such as a drug reaction, a transfusion reaction, DVT, or pulmonary embolus.

But obviously the decision that is made is not made on the basis of any one of those parameters, but the integration of those parameters into an overall clinical probability that will probably include 6 or 8 different variables from the clinicians perspective.

DR. DELLINGER: From a Day 3 standpoint, it would be great if we could totally rely on the negative culture for Gram-negative organisms, and that would be wonderful if we could just use that isolated from other clinical factors.

But we really can't. The essence of it is that on day three, or when our culture results are back, we use that as a very important piece of information to decide that we don't need to be concerned about GRAM-negative infection.

But there are many patients in which just a negative culture is not enough based on the whole clinical picture, and in that circumstance there would be another significant percentage of those patients that combined with the negative culture and the negative EAA that would give us the comfort to say that we are not dealing with GRAM-negative infections.

And there are likely even to be -- I mean, there is likely to be -- I mean, it says rule out, but there are going to be some patients where the total clinical picture would be that the clinician, even with the negative assay and the negative cultures, might still decide to continue antibody coverage.

DR. RELLER: I understand everything that has been said, and that cultures are not enough to rule out GRAM-negative infection, and the presence of GRAM-negative infection. Is this test enough, and what does enough lead to?

Is it enough to stop the antimicrobial therapy directed at the GRAM-negative? Is it enough to not get a CT Scan? I mean, enough to take what specific action? What does it add to what we have in terms of enabling, or either doing something, or not doing something specifically?

DR. DELLINGER: I think, and I am going to sort of repeat what I said a little bit, but I think that is a great question, because we make a decision at the bedside based on 9 or 10 pieces of information that we think are all important.

This would be one more piece of information that would be important in the decision making, and when we make the decision about continuing antibiotics for GRAM-negative infection or negative assay.

But I don't think we ever make the decision about continuing coverage, or stopping coverage, based on any one single variable.

DR. SOLOMKIN: But how much weight would you give this? Like if you had a negative CT Scan, you would give that substantial weight. Would you give this as much weight as, for example, a negative CT Scan?

DR. DELLINGER: Joe, I don't know the answer to that. In fact, if you look at things like pulmonary embolism, where we use an amalgamation of accepted lab tests, and other variables, to decide whether we do or don't have PE.

But yet any one single one of those would not be enough. You know, ELISA D dimer, is that useful? Yes. Other pieces of information are useful, and I don't know whether any of the statistical people or John could actually put a measure.

But I imagine it is going to vary from patient to patient how important that piece of information is for that particular patient. But I think it would be a piece of information that would be important, and varying in importance from patient to patient.

DR. WALKER: Can I just make one comment on that as well? And that is that if we go back to the question about a CAT Scan, most of those other investigations tend to be in the direction to prove an infection, and not to rule out an infection.

And I think that there is this subtle change in thinking in this that in fact -- and again as both John and Phil have said -- that any clinical diagnosis is made up of a number of different bits of information, which are weighted differently and often in different patients.

At the moment, we have very little information that in any way pushes us away from the diagnosis of an infection. So we believe two parts of this, and the first is that with respect to the presence of GRAM-negative infection, this could give you some relatively early information, which makes at a level of 94 percent the actual, eventual diagnosis of an infection unlikely.

And if given in the fact of that, it may alter one's pursuance of a diagnostic, particularly in the area of a GRAM-negative. But clearly in every situation it is only one piece of information.

On the other hand, it is new information, and it is information in a timely manner. And it is information that I think we can say has been pretty rigorously evaluated in the clinical situation.

And I would reiterate that this clinical test -- I'm sorry, this clinical study, is a very heterogeneous group. It is the kind of people that we see in the intensive care unit with a number of disease processes going go.

The reason for admission as you have seen in the PMA were varied. There is very little screening of these patients coming in. So this test has actually proved robust in a situation where there are multiple potentials for the presence of GRAM-negative infection.

DR. RELLER: But to follow up on this rule out emphasis that has been made. I mean, the sensitivity of this test is in the order of 80 percent as portrayed. Let's not rule out with the sensitivity in the prevalence of the negative predictive value in the 90 to 94 percent.

I mean, it is highly dependent upon the prevalence of the entity that one is seeking to rule out. So that if you look at 90 or 94 percent, you know, that gives you one impression. And 80 percent, is that sensitivity sufficient to exclude an entity.

DR. WALKER: I understand your question, and I think that we have to take those statistics into this group of patients, and this group is a very complex group of patients and it is very difficult to make a clear diagnosis.

I think we have established some of that. With respect to the certainty, I think if we look at it from -- we can look at that from a number of different points of view. The clinical suspicion of infection actually results in 92 percent of those patients being false-positives.

And the ability to have with the assay then is to convert some of those to true negatives, and it is unlikely that you are going to be able to convert them at a hundred percent negative predictive value, and we recognize that.

On the other hand, it is a piece of information that converts a significant number of those people to perhaps a differential diagnosis which is altered that may result.

And I think that part of our thrust in doing this is the current techniques that we have of a patient changing a status, and putting those patients on antibiotics, taking cultures, and waiting for three days, has been proven very effective in the last 20 years.

We really have not made much impact on this overall conundrum. So I think the thing that I would underline is that this is one piece of information that adds confidence in this situation, and certainly we have seen in the practice is that while 80 percent of these patients are on antibiotics on day one in this study, 80 percent of those patients are also on antibiotics on days 3, 4, 5, 6, and 7.

So the current practice would seem to be that despite the fact that a negative culture comes back, there actually hasn't been an acting upon that. But I think both John and Phil addressed the point that the adjunctive piece of information you get is in the presence of a culture.

Also, the CEC adjudication did not just look at cultures. It looked at the entire clinical picture, and made a decision on whether that patient actually had a Gram-negative infection going on. So that is the added piece of information.

It just added confidence in that ability to perhaps not only confirm the diagnosis, but in fact alter therapy in conjunction with the cultures.

CHAIRMAN WILSON: We have time for three more questions. First it will be Dr. Danner, and then Dr. Janosky, and then Dr. Ing.

DR. DANNER: I think I am having a problem with your number, the negative predictive value number. I am going to call it 91 percent and not 94 percent.

But that number, how clinically meaningful is that number given the definition that it is based upon? Your numbers are all calculated based on the assumption that a negative culture, a culture that doesn't grow, means no infection.

That's not true. That's not true in any ICU that I have ever practiced in. A negative culture doesn't mean no infection. There are infected patients in ICUs who are on antibiotics, and they come to you from the floor on antibiotics, and you are unable to recover any organism.

So that value of 91 percent, though it is a correct number based on the way that you define what an infection was, I question the clinical reality of that number, in terms of whether someone is really infected.

Getting back to something that Phil said, Phil said, well, on day three, if I get a negative culture, and then I have this test that is negative, then that gives me more confidence to act clinically based on that because I have two pieces of information.

The first piece of information though, your negative predictive value, is based on the fact that the culture -- that the whole calculation is confounded, and they are not independent of each other.

DR. WALKER: I understand your question. In our interactive discussions with the FDA, it was clear for all the reasons that we have talked about today, and that is the importance of diagnostic information in these patients who are so critically ill, that our test essentially had to be related to cultures.

DR. DANNER: Now, I understand the reason for using that as your standard, but you have a tarnished gold standard, and to take that number of 91 percent and feel that you now have a great level of confidence that a negative test, your test, means anything, is I think clinically dangerous.

DR. WALKER: I have two issues that I would say on that. Number One is that the patients who are not -- did not have cultures, they were reviewed not simply for that culture at that point.

These patients were reviewed for their entire stay within the intensive care unit. So in those patients, not on one day, and not in any site, did they ever have any positive culture of any kind.

So I understand your quandary. The CEC looked very carefully at the entire spectrum of the culture reports over the entire time. Their adjudication was the presence or absence of infection on day one based on the temporal relationship between the cultures being taken and the overall process.

DR. DANNER: I understand how all of that was done. You still can't get around the fact that your number of 91 percent or 94 percent, or whatever you want to call it, is based on assuming that somebody with a negative culture has no infection, and that is not true.

DR. MARSHALL: I would like to comment on that, because what you are touching on is exactly the rationale for establishing a clinical evaluation committee.

You are right that you are in a Catch-22 situation, and that if you define the presence of infection by cultures alone, then how do you deal with the possibility that you may have an infection that is culture negative.

We had a CEC that reviewed all of those cases, and this is an expert group of people with expertise in infection in critically patients. Their adjudication was that something like 40 percent of those patients that were adjudicated by CDC criteria as being infection, in fact when you look at the whole clinical package, were not infected.

So the reality is that as much as the fear is always there, that that hundred percent of patients with suspicion of infection actually harbor infections, but for some reason the organism wasn't isolated.

And they are antibiotics and the wrong samples are taken, and specimens were lost, and when an expert group of people with knowledge of the diagnosis and management of infection in the ICU review the data, their conclusion was that the CDC criteria overestimated, rather than underestimated, the cultures.

DR. DANNER: Yes, but it is still only looking at that subgroup with positive cultures at any site. So you still have a problem of the people without positive cultures weren't even part of that evaluation.

So that number -- and I just want to point out to everyone on the committee that number of 91 percent is a very soft number, and in terms of clinical relevance of that number, the true number is something less than that.

I don't know how much less than that that it is, but it is less than that.

DR. WALKER: From a statistical point of view, the use of the negative predictive value is a challenge in this because of the definitions that we were forced to accept, which were the definitions of infection. So I understand what you are suggesting.

And that the negative predictive value, which would be the normal way of looking at that, is challenged in this situation for a number of different reasons.

To reassure you from your point, I would say the following, and that is that if all of our negative patients came from that group that had negative cultures, then your comment would have more validity, and that is not the case.

In fact, a significant number, more than half of our patients, actually came from the group that had negative EAA, but they had positive cultures, in that group that was reviewed with the positive cultures. So I don't believe think that we --

DR. DANNER: I don't think that actually answers the question. The other thing is that in terms of our clinical data, a lot of your in vitro testing is interesting, but you in fact need to show us the stratification of the clinical data based on people on antibiotics, and off antibiotics at the time that the culture and that your test was done.

And in terms of the false positives that you are getting, what I would like to know is people who clearly had fungal infection, or GRAM-positive infection, and that is all that you could identify in them.

Therefore, i.e., people with just clear cut Staph aureus line infection, or pneumococcal pneumonias, things where Gram-negatives were not involved, and how did your tests perform in those patients.

And how many of them were falsely positive, who had no GRAM-negative infection, and I have not seen that kind of -- you know, a lot of stuff has been geared towards the in vivo testing, and I have not seen a lot of the stratification from the clinical trial that in fact the committee needs to be able to evaluate how this test performs clinically.

DR. WALKER: Those are actually a couple of questions. Could you just repeat the first of the questions. The in vivo, we can present some more of the data if you would like, but I would like to know specifically what you would like.

And with respect to the false positives, those are false positives in the structure in which we have been asked to look at this test. Those are false positives between the relationship between endotoxin being elevated and GRAM-negative. They are not false positive endotoxin elevations.

DR. DANNER: I don't think you actually know that. You can say that you know that, but you don't in fact know that because there is not a gold standard in relationship to endotoxcemia.

So you can't in fact say that those are -- that those people really have circulating endotoxin, and that it is not something else that is turning your test on.

CHAIRMAN WILSON: Oh, I think we had very good evidence. If you have looked at our publication, The Journal of Immunological Methods, and I think in the presentation from Dr. Romaschin today, two things.

Number One is that I think this assay is highly specific for endotoxin. You are looking at very clear -- both studies, as to what organisms have response to, and we have done obviously the best that we can in comparison to other tests that are out there, like the LAL test.

DR. DANNER: Right. And none of those tests -- in none of those tests can you be sure that what the test is measuring is endotoxin, the actual physical molecule in the blood.

DR. WALKER: That is clearly true with the LAL. It cross-reacts with GRAM-positive and with fungal. We have clear evidence that ours does not do that, and that has been published in the --

DR. DANNER: Yes, but in the clinical setting there are other things that could be activating those, activating the cells, and I don't think --

DR. WALKER: But the specificity of the assay rests upon the specificity of the antibody,a nd that is a very well characterized antibody that is very specifically related to the Lipid A portion.

It is very highly conserved in every one of those organisms as we have identified, and so I don't think that we are challenged because there isn't a gold standard either of infection or endotoxin for us to compare ourselves to.

DR. DANNER: Just show the data. Show the data for people with GRAM-positives in the blood, and people with Candida in the blood, and how the data for how your test perform. Just show the data.

CHAIRMAN WILSON: I would like to say that at this point that we need to move on for other questions. Dr. Janosky, please.

DR. JANOSKY: The question is more likely appropriate for Dr. Marshall. I might be incorrect, but let's start there. At this point, I want to gather some more information. I am very interested in the issue of prevalence, and how different prevalence levels will affect what you are reporting as your outcomes.

Do you have data to show either those values by the sites or by patient characteristics; and if you do, I would like to see that, please.

DR. MARSHALL: The data for endotoxin, per se?

DR. JANOSKY: Data for your calculation of a negative predictive value based on different prevalent values. And you could look at those based on either your three largest sites, or you could look at that based on patient characteristics, and I did not see those data presented. So I would like to see those, please.

DR. MARSHALL: I don't have those data off the top of my head. You are right, that it is going to vary, and it is only going to be valid for the sites.

I can comment with some sense of modest embarrassment that there was one site that seemed to have more -- we seemed to have missed more cases, and that was in fact the site that I come from. I think 5 of the 8 missed cases were in fact from the site that I was at.

One of the sites had no missed cases, with roughly comparable prevalence of GRAM-negative infection. But I don't actually have the specific numbers for you.

DR. JANOSKY: Well, what were the ranges of prevalence? We can talk about this a little later, but this is one of the issues that I am very concerned about. So perhaps it will give you some prep time to gather some information.

DR. MARSHALL: I would have to actually review the numbers to give you those. You want prevalence of GRAM-negative infection by site?

DR. JANOSKY: Exactly, and you have three recent sites, and then you also have patient characteristics, and if you could give me the prevalent values; and then what are the NPVs for those.

DR. MARSHALL: And patient characteristics, you are talking about demographics, the severity --

DR. JANOSKY: Well, we don't have this in our packet here, but I did see a presentation up there that showed the location being one of the variables, and lung was the largest, I think?

DR. MARSHALL: Yes, that's right.

DR. JANOSKY: As well as some of the other variables.

DR. MARSHALL: And you would like to see the location broken down by site?

DR. JANOSKY: At least for the largest locations to get those NPV values, because I am very interested to see what the effect would be on that.

DR. MARSHALL: All right.

DR. JANOSKY: I don't want to take up time now.

CHAIRMAN WILSON: Dr. Ng.

DR. NG: I have a comment and a question. The comment that I would like to make and perhaps hear the rebuttal in the discussion phase, but looking at your data and your analysis, 8 percent of your patients ultimately had confirmed GRAM-negative infection.

That tells me up front that if I were looking at your study group that I would have a pre-test probability that 92 percent, the flip side, lack GRAM-negative infection.

The negative predictive value of your test is 91 percent, and another way to state that -- and this gets to Dr. Reller's question, and I am not sure how to use this test in a clinical setting or how it affects patient management.

That although the goal of your test is to rule out disease, clinicians want to kind of think about both the sensitivity and the specificity. So when I go ahead and calculate a likelihood ratio, which is sensitivity over one minus specificity, and I am considering the odds that the patient that I am looking at has the disease, the likelihood ratio is no greater than 1 to 1.2.

In other words, if someone comes up with one to one odds of having a GRAM-negative infection, and I do the test, I end up with posterior odds of 1 to 1.2, and that doesn't seem to get me too far. So I would like to hear your comments on that type of analysis.

DR. WALKER: I would just make one comment about the numbers, and just so you are clear that if you are making the comparison between CDC numbers, the incident of infection in that group was 13 percent, and are a negative predictive value of 91 percent, and the other group 92 percent and 94 percent.

And I think I would ask Dr. Willan to make a comment about the challenge of using ordinary statistics, including likelihood ratio, in a group where we have a significant number of false positives, because that is the problem that we run into.

CHAIRMAN WILSON: Could you identify yourself, please.

DR. WILLAN: My name is Andy Willan, and I am a Professor of Biostatistics at McMaster University, in Canada. I think what we have done in this data analysis is concentrate on NPV and not on regular ratio -- the positive test is not going to help us rule anything in.

So things like concentration ratio would depend considerably on the specificity of the test as well as a sensitivity. So we don't expect this test to have a good likely ratio for a positive test.

DR. NG: And this is just my final comment, but I do have a question, but my final comment is that I am left with a pretest probability of anywhere from 83 to 87, to maybe 92 percent, and this test gets me to 91 percent.

But my question for the group is the precision of your assay is about 15 percent, plus or minus 15 percent. Would you please comment on how your results would be affected if you factored that in, in terms of your true negatives?

DR. WALKER: We have a slide on precision, and I don't suspect that is going to particularly answer your question. So I think we will have to provide you with that answer.

Just to reiterate the statistics, which we don't disagree with. We have looked at this quite carefully. And the issue is really a degree of confidence, and I think that is really what we are pointed at, and that clinical suspicion has a pretest probability of infection of a hundred percent.

These patients are concerned to be a hundred percent. Most of them are not infected. At the moment, everyone treats them as if they are infected, because they have nothing that gives them any confidence in that three days that they are not infected.

So the issue of comfort or confidence in this is related to the fact that a negative EAA is associated with a low incidence of GRAM-negative infection.

So out of those patients that just by definition clinically you have assigned them to have a suspicion of being infected, well, only 8 percent of them on CDC, or 13 percent -- I'm sorry, 87 percent with CDC, in fact are going to have an infection.

And there is nothing at the moment that tells you which of those patients, and there is no confidence that we have. Our test does convert a significant portion of those patients from essentially a clinical false-positive to a probable or to a true negative.

So that is the advantage of it, and at the moment there is nothing else out there that in any way adds that degree of information in that period of time.

So while we recognize that, we recognize that the likelihood ratio is a challenge given the fact that there is so much noise of endotoxin in the background, and that is very well accepted by most investigators.

So that the likelihood ratio is clearly going to be affected from a numerical point of view by the specificity of only 33 percent. So while I recognize what you are saying, and we have grappled with that with respect to how we would express these results.

And I think that Dr. Danner is saying exactly the same thing. How do you express these results in a way that are going to be useful for the clinician.

I do think it is clear that the results of the MEDIC trial have shown that in a very diverse group of patients from a number of different centers in the world, which on paper represent the kind of challenge we get on a regular basis with a high mortality rate of 28 or 30 percent.

And with the incredible use of antibiotics in these patients, and the numbers of cultures -- and I would go back to Dr. Danner's point. And that is that every single one of these patients was cultured on a regular basis.

And the cultures, although we have mandated on day one, and we have a slide that shows that even during the entire course of this observational study, where we in fact didn't direct them, these patients were cultured multiple times every day.

So I think it is clear that there is a diagnostic dilemma, and I think that we believe that our assay is new information, and it is important information, and it is timely information.

And it is a piece of information that has to be taken into account with all of the other aspects that are being used in patient management, and that is what we would propose.

CHAIRMAN WILSON: Okay. At this point, I would like to ask the panel to hold any further questions until the open committee discussions this afternoon. I would like to have the FDA give their presentation now.

All right. While they are setting up, let's go ahead and take about a five minute break here.

(Whereupon, at 11:39 a.m., a recess was taken, and the meeting was resumed at 11:46 p.m.)

CHAIRMAN WILSON: Okay. At this time, we would like to go on with the FDA presentation. Again, I would like to ask the panel members to hold any questions until after the two presentations have been completed.

The first presentation on EAA performance characteristics will be given by Marian Heyliger who is the senior scientific reviewer for the Bacteriology Devices Branch.

MS. HEYLIGER: Thank you, Mr. Chairman. Good morning members of the panel. We are in agreement with the facts are presented by the sponsor. I want to remind you that the PMA came in as an expedited review, but we are still reviewing the PME. It is still currently under review.

But we brought this application to you to seek some input form you in order for you to help us determine the assay's role in clinical lab diagnosis. So we are going to take a look at the assay from a slightly different perspective.

I will touch very briefly on the following topics, which are the spectrum of sepsis, lab diagnosis, medical trial results, the described population, assay limitations, and the conclusion.

The endotoxin activity assay has as its intended use the measurement of endotoxin activity in human whole blood as an aid in ruling out the presence of GRAM-negative infection in ICU patients suspected of infection.

But in the past, however, GRAM-negative organisms were the most common blood culture isolates against robotically E. coli Klebsiella pneumoniae.

However, the spectrum of sepsis is changing, and the theory perhaps that is circulating endotoxin is responsible for a lot of the morbidity and mortality of sepsis probably is being challenged by the fact that many of the organisms now being isolated are GRAM-positive organisms, like Staph Aureus and enterococcus, and coagulase-negative staphylococcus.

And in addition we see Candida and Fungi. This information comes from the National Surveillance System in Richmond, and from CDC in Atlanta. Now, identifying patients with sepsis from clinical criteria can be difficult, and so making a lab diagnosis perhaps is an important adjunct.

Traditionally, blood cultures have been regarded as the gold standard for establishing the presence of bacteremia, but we all know that its value is questionable, and that true sensitivity cannot sometimes be determined.

There is a delay in results from blood culture, and sometimes about 24 hours. The sponsor has proposed that the endotoxin activity assay as a rapid diagnostic, offering an advance to aid the clinician in diagnosis, and giving timely results of less than four hours.

The pivotal study done by the sponsor was the MEDIC study. For each patient in the MEDIC study, there was an order for one or more diagnostic cultures.

Let's look at the one study culture results which you have seen before, and so I will probably go through it very quickly. There were 73 patients with GRAM-negative infection, and 54 of them were determined to have GRAM-negative growth as defined by the CDC criteria.

And 33 were determined to have GRAM-negative growth as defined by the next level, which was the CEC adjudication committee. There was disagreement with standard infection definitions as provided by the CDC criteria.

And, of course, there was difficulty in determining the patient's infection status. If we look now at the second slide, here we have the results using the CDC classification for GRAM-negative infection.

Out of a total of 408 patients, that was the endotoxin patient level. If we look at the top line, the 120 patients out of 408 had a negative endotoxic activity value.

Of those, 117 patients had no GRAM-negative infection, but there were 11 that fit the CDC criteria for GRAM-negative infection, and these probably could be regarded as the false-negatives.

There is a presumption here that a negative endotoxin activity value correlates with the absence of GRAM-negative infection. If you look at the row below, the second row, there the endotoxin activity value is over .4, which is regarded as a positive EAA value.

There we had 280 patients with that result, of which 43 fit the CDC criteria for GRAM-negative infection. That left a total of 237 patients who had no GRAM-negative infection, but a positive EAA result. These can be regarded as the false positives.

The negative predictive value, as we have mentioned before, the negative predictive value is 91 percent. If we look at the next table, which shows us these 408 patients, now we are determining these results using the CEC classification.

It is the same 408 patients. We have now 120 of them showing no GRAM-negative infection, and 8 of them with GRAM-negative infection, and all 120 had EAA values less than .4.

So we see here that our false negative has now dropped from 11 to 8 when we look at the CEC classification, as opposed to the CDC. If we look at the second row where you have a positive EAA value over .4, we see of the 280 patients, that 255 had no GRAM-negative infection, and 25 had GRAM-negative infection.

Our false positive rate now is 255. So we see a decrease in the false-negative rate from 11 to 8, but we also see an increase in negative predictive value to 94 percent.

Now, let's just consider something with a false-positive population. The test itself showed a sensitivity of 80 percent based on GRAM-negative infection, and the previous table showed us that the false-positive cases were 237 by CDC criteria, and 255 by the clinical evaluation committee.

The false positive rate was not reviewed by the sponsor due to a lack of specificity of endotoxin production. It is well documented that endotoxin could arise from sources other than GRAM-negative infection.

But we would like to know that should the false-positive results be addressed should they be included in the assay evaluation. Do they reflect the assay's non-specificity.

The next slide shows us the false negative population. This is a population that we need to look at, bearing in mind that one of the key parameters of the assay is the negative predictive value.

The false negative population consisted of 11 cases. It is broken down into two slides. The first slide is used for the first five cases, and the second slide will cover the rest.

If we look at the first slide, we see that infection was determined from various sites; lung, blood and urine, CNS. A variety of organisms grew; pseudomonas, klebsiella,and serratia.

We know, too, that the endotoxin activity value on day one in every instance was less than .4. As regards to mortality, most patients lived, and then when we come to the description, we see that in four cases that both the CEC and the CDC agree that the result was a false-negative.

In case 1, and case 1-A and 1-B, it was from the same patient, brain serratia, and from a lung specimen, and pseudomonas from blood and urine; and the blood culture was GRAM-negative, and the EAA values remained negative, and this was regarded as a true-false negative.

When you come to 2-A, which is the third case, this patient, with serratia from the lung, the EAA value was less than .4, but the CEC and CDC disagreed here because the blood culture was negative, but the sputum growth was light.

And so the CDC determined that the person did have infection, but the CEC thought that it was colonization. When we look at Case 3, this was a case of klebsiella meningitis, and so that was clearly a false negative.

Case 4 was an endotracheal aspirate, and that was mis-classified. It did not meet CDC criteria for pneumonia and the blood culture was negative, and so on.

The last one was a false-negative, and if you look at the next slide -- and I am not going to go into these in any detail. But again the picture is the same. There was disagreement in three cases, and false negativity in two, and negative EAA values for all, and growth on all cultures.

The endotoxin infection diagnosis is clearly evident. Now, if we look at the limitations of the endotoxin activity assay, we could probably explain that from the fact that there might be a non-hematogenous site of infection so that endotoxin is not detected.

And bacterial probably might not be shared into the blood flow. It could be a remote site of infection, with no circulating endotoxin. Perhaps endotoxin has not achieved access to the systemic circulation.

There might be positive bacterial cultures in the absence of endotoxsemia probably due to colonization, since colonized activity doesn't always affect the systemic effects of infection. Of course, there might be other contributing factors to explain the false-negative population.

Now, in conclusion, what I want to say is that the primary objective of the endotoxin activity assay was its reliability to exclude the diagnosis of GRAM-negative infection in critically ill patients with suspected infection admitted to the ICU.

Only day one study entry data was used in the assessment of the endotoxin activities performance, and we wonder could infection outcomes be better determined beyond day one.

And the NPV of 91 percent or 94 percent as demonstrated in this study, could that indicate a role for this assay in clinical lab diagnosis. And this assay is currently under review by members of the department as indicated.

I would now like to introduce the next speaker, our statistician, Mr. John Dawson.

MR. DAWSON: Thank you, Marian, and thank you, Dr. Wilson, and members of the panel for the opportunity to present the FDA's statistical perspective on this application.

Much of what I plan to say has already been discussed; Dr. Reller bringing up the point about the negative predictive value being so close to prevalence, and Dr. Danner talking about the gold standard and whether there is one here, and Dr. Eg's likelihood calculation.

The problem that we have with the negative predictive value, first of all, is that it does require a gold standard for unbiased destination. It is a function of sensitivity and specificity.

And by gold standard that means that you have got to have a really reliable way of avoiding both false negatives and false positives, and diagnosing a disease condition, and it is questionable as to whether that exists.

If it does exist and we take the negative predictive value at face value, that 94 percent, the confidence interval on that 94 percent includes prevalence, and that shows up in one of the sponsor's slides, and it was a calculation that I duplicated.

Sample size has a role in this, and had the sample size been something in excess of 2500 instead of 408, the confidence interval on that 94 percent would have had a lower limit that went above the 92 percent prevalence.

And in which case you would then be back to the likelihood calculation, and you would have to ask yourself does that two percent margin over prevalence really constitute clinically utility.

And I say that even understanding and appreciating the sponsor's point of view that nobody is going to focus only on that one particular test. But the fact that the confidence interval includes prevalence means that it is no better than a random test, which literally means that you could do as well with a table of random numbers, as with the assay result.

In light of the question as to whether there really is a sufficient gold standard, what I want to suggest -- and I am just going to go to the next to the last slide in my presentation.

And basically what I am suggesting is that these terms of art -- sensitivity, specificity, and predictive value -- we need to respect the fact that those are probabilities, and that imposes a discipline as to what kind of calculations you can do and apply those terms to.

And if you don't have a gold standard, then it is a misuse of that terminology, and it is misleading to the user if that is present in the labeling.

But a simple way around that is to replace the statement that the sponsor makes, "A negative EAA result is consistent with the absence of the disease in 94 percent of the patients."

And to replace that with a statement such as, "A negative EAA result means there is a 94 percent probability that this case would be found disease negative by CDC criteria or by CEC, plus clinical adjudication."

So basically what that 94 percent does that is of some use I think is that it indicates what the relationship would be between a patient that the test is applied to and what the diagnosis would have been, or the disease status determination, if that patient had been in the study.

Now that is basically predicting the outcome of study truth when you are looking at a given patient, rather than saying that the study has really give us a confident way of assessing the likelihood of disease.

Lastly, I just want to point out to Dr. Janosky that the sponsor recently provided some site-by-site, two-by-two tables, which I have looked at, and I did look at the negative predictive value, and it was consistently in the mid-to-upper 90s across sites.

What I didn't do, and what I think you want to do, is to compare that site-by-site with prevalence. But they did provide that, and it was kind of very recent. Thank you.

CHAIRMAN WILSON: Thank you. Do any of the panel members have questions for the two FDAers? Dr. Nachamkin.

DR. NACHAMKIN: So that we can get back to the study design, and whether one can have confidence in the numbers presented by the sponsor. And, Mr. Dawson, the comment that you make in terms of replacing negative predictive value with this other comment, can you actually say with any confidence that this test will rule out disease in 94 percent, when they had such a small sample size?

In fact, the confidence interval goes down to about 84 percent. So isn't that misleading to say that we are confident in 94 percent, where in fact it may be as low as missing 15 or 20 percent of the patients?

MR. DAWSON: Right. We normally look at an effectiveness measure, in terms of its lower confidence limit. And taking the 94 as the point estimate, and calculating the 95 percent by the binomial confidence interval, the lower limit I got was 88 percent.

So we would basically look at that and say that this has shown something in the neighborhood of 88 percent or better. But it definitely could be as low as 88 percent.

CHAIRMAN WILSON: Dr. Charache.

DR. CHARACHE: I'm coming back to Dr. Dawson's comment about commenting that a negative EAA result means that there is a 94 percent probability that it would be found disease free.

And reminding ourselves that it doesn't mean disease free. It means negative culture, and all the problems with being able to culture the side of pathology or interpret a pulmonary culture if that is the side of pathology.

Or to know if the cultures were taken when the patient was on antibiotics or not on antibiotics. I think we have to be very careful about talking about this in terms of prediction of disease, and simply say prediction of culture negative, and we don't know the conditions under which the cultures were taken.

CHAIRMAN WILSON: Dr. Baron.

DR. BARON: I have a question for Marian. When you looked at the exact EAA values of the false negative population, it is sort of striking to me that many of them were between .3 and .4.

And I am just wondering that when you saw all of the data, which I did not see, is there perhaps an equivocal zone on this result, that if the sponsors were to lower their positive threshold that we would not see these false negative patients?

MS. HEYLIGER: Well, I believe that in the equivocal study that the sponsor did actually lower their cutoff to .3, but I think when they actually did the medical trial that they used .4, because there is no equivocal zone in this assay.

MR. DAWSON: Let me just introduce a word of caution about that, after the fact changing a cutoff. We are often tempted to do that because we can see better performance if we change the cutoff.

But what that tends to do is to give you an unvalidated cutoff, and tends to give you an overly optimistic picture of performance. So we are very careful about that kind of adjustment after the fact.

CHAIRMAN WILSON: Dr. Charache.

DR. CHARACHE: I wondered also if the FDA had had the opportunity to look at some patients who were not culture positive that had the same clinical presentation to see how the criteria of the clinical assessment panel would have been, but they had thought that the patients did or did not have infection, if that data was available to you.

MS. HEYLIGER: We have not reviewed that data. It is important to remember that the claim that the sponsor is making requires that we only review the data from the study, day one, from day one of the study.

DR. CHARACHE: And I am wondering about day one if there was an opportunity to see whether patients -- well, how the clinical evaluation committee would have judged them, because that is a very important criteria as to whether patients were considered to be false negatives, or too negatives.

MS. HEYLIGER: Right. And all I have on that is just from the 11 pieces that I have presented on the slide. That was the only data that I presented.

CHAIRMAN WILSON: Dr. Baron.

DR. BARON: But you asked the question could infection outcomes be better determined beyond day one, and I believe I remember from the study protocol that they tested EAA every day for at least a week.

MS. HEYLIGER: Right, but this data was not -- was not included for the claim, because the claim for the study is ruling out GRAM-negative infection, but it is only -- but the data is only to be reviewed for day one of the study.

And that is why we asked the question; whether in fact you could get better outcomes if you looked at data from other days of the study. Perhaps the manufacturer has some of this data, but it is the data that they want us to review for the claim is day one of the study.

DR. BARON: Yes, but they are making their decision point on day three.

CHAIRMAN WILSON: Dr. Durack, you are next.

DR. DURACK: Mr. Dawson, I wonder if you could comment from a statistical point of view on the possible value or non-value of repeating the test, and what if a negative test as we understand it now were repeated on day two and day three, or twice in one day? Any comment?

MR. DAWSON: Not from a statistical point of view. It is often something that we see, that a protocol will call for that, and that if you get a discrepant result between two tests, one of which is the accepted standard, then you need to repeat it.

I don't think that was an element of this study.

DR. DURACK: I am just saying if it were done.

MR. DAWSON: Okay. If it were done, then -- well, what is the question?

DR. DURACK: Would you get increasing predictive value by repeating or negative predictive value?

MR. DAWSON: I would assume somehow with more information that you could get more out of it. I am not sure right off the bat what that would be.

CHAIRMAN WILSON: Dr. Nachamkin.

DR. NACHAMKIN: Yes. Just to clarify. I don't quite understand why we only have to consider the day one data, when in fact all the presentations made by the sponsor said that this test isn't a day one test. This is a day one and day three test.

MS. HEYLIGER: Well, initially when the sponsor presented -- can I state that?

DR. GAFFEY: Dr. Claudia Gaffey with the Division. The culture was taken on day one, and the decision -- the result of the question is that it comes on day three. We were asked to review the data on day one.

The presentation that was shown today was not actually included in the submission, the graph showing the day one, day two, and day three. We knew that the results of the question would come after day three. However, these are the cultures that were present that were taken on day one.

DR. NACHAMKIN: But the way the test was presented was that this was not a test just solely to be used to rule out infection. It is a piece of information to be used with other variables over that course of time.

And it was specifically repeated that after a couple of days, if you received negative cultures, that that in combination with negative EAA tests, would better help to rule out infection. So again we are getting conflicting instructions here as to what to consider.

DR. GAFFEY: Well, on day two, there were other diagnostic tests that would or could have probably done it. But that is the way the review was done, and that was the way that we were directed to proceed. I agree with you.

CHAIRMAN WILSON: Dr. Danner.

DR. DANNER: In terms of the false negatives, the organisms -- I am wondering if the organisms shown here are the same organisms that you see in the true positives?

You know, there is a lot of pseudomonas in here and serratia, and things, and is there a random distribution?

MS. HEYLIGER: Well, unfortunately, I can't answer that question because we did not get the data from the positive population. The only data that I have been provided with is the data from the false negative population.

Remember that the manufacturer was not keen on reviewing the false positives because of a lack of specificity.

DR. DANNER: Well, I think that is important because as I think that kind of came out in some of the presentations, antibody specificity and detection, and things like that, may differ across different species which in fact have different endotoxins, and not one molecule, but many molecules.

And might there be some types of GRAM-negatives that would be missed more often than other types? Is there someone from the company that can answer that?

CHAIRMAN WILSON: Yes. Would someone from the sponsor like to comment on that? Dr. Walker.

DR. WALKER: Is it possible to have another one of our slides shown.

DR. DANNER: Maybe if you could just say what the percentages of serratia and pseudomonas is, and --

DR. WALKER: Well, first of all, Dr. Danner, we are dealing with a population of 33 patients, of which eight are in one category, and 25 are in the other.

And we have a slide that shows exactly that, and it answers your question. And specifically that the distribution is the same in the two groups, both the true positives, and in all of the ones that are infected.

DR. DANNER: So there is nota any pseudomonas in the true positives?

DR. WALKER: Yes. So there is not one organism that would appear that we are repeating. Am I at liberty to answer one of the other questions that was asked?

CHAIRMAN WILSON: Go ahead.

DR. WALKER: If we had the opportunity to show a slide, because it may throw some light on what we are discussing, and the issue that was brought up previously, and that is that we do have a slide that was provided to the FDA.

But given the challenge in the last month on getting documents across borders, and through Federal groups, it is not surprising that we have not been able to challenge it here.

But there is a slide that actually shows a group of patients that are dichotamized solely based on endotoxin assay. I mean, it is relatively interesting, and that is -- or in other words, it goes back to your question as to what do these patient populations look like, endotoxin positive or endotoxin negative.

And is there a discriminating function related to what we are doing, and I would love to show you this slide, because in fact the essence of the slide shows that the populations are virtually identical.

And it is very challenging to separate those patients on any of the normal parameters that we use, but the only thing that is very different in that in those that are endotoxin negative have a very low incidence of GRAM-negative infection.

And those that are positive have a much higher incidence of GRAM-negative infection, in keeping with our sensitivity. So, 80 percent of the patients are in the category of greater than .4 have a GRAM-negative infection.

DR. DANNER: Well, they are culture positive.

DR. WALKER: Yes.

DR. DANNER: They didn't grow in your cultures --

DR. WALKER: Yes, and I go back to your comment on that, because it is a very valid comment, and it is the same as the other comments that have been changed. And I think in Mr. Dawson's discussion on challenges of negative predictive value, and applying that to this particular case, I think have some merit to them.

And the issue of what we really have shown is agreement. I mean, we have shown agreement more than we have probably shown negative predictive value. We have shown agreement in the confines of the protocol that we put forward, and the protocol where the FDA was very anxious that we link endotoxin and infection.

And so I think that the points are true, because using negative predictive value in this situation, and as Mr. Dawson said, requires a gold standard, but it tends to go in both directions.

Whereas, we at this point cannot say that more information is added to those people with infection with a GRAM-positive. I'm sorry, with a positive endotoxin assay.

DR. NACHAMKIN: Could I just ask one question?

CHAIRMAN WILSON: Yes, go ahead.

DR. NACHAMKIN: You group these patients as ICU patients, and again the data that we got in our folders is very limited, in terms of patient data. What kind of ICUs were these? Was this a medical ICU, a surgical ICU? What is the mix?

DR. DELLINGER: There was -- well, most ICUs tend to be mixed certainly in the United States, or -- well, I'm sorry, in -- Well, a medical ICU would be called --

DR. WALKER: The Medical ICU at Abrahams in Denver, which I think would be called Medical ICU.

MR. DELLINGER: Medical.

CHAIRMAN WILSON: Could you come to the microphone, please.

DR. DELLINGER: The intensive care units at Chicago were medical, but many -- you know, maybe five percent of our patients go to surgery, and if it is not cardiovascular surgery, then tend to come back to us.

So there is a population of surgical patients, but it is certainly that the great predominance are medical. And I think the same thing for Brown, but I can't --

DR. NACHAMKIN: So basically your claims further narrow the population at risk to patients in a medical ICU. It doesn't include patients in other ICU settings?

DR. DELLINGER: No, that is just U.S. I think that the predominance of the units in the study would have been some surgical and mixed med surg.

DR. WALKER: Yes, the vast majority of the patients who were admitted came from mixed units, and that includes the unit in Brussels, and the units at the Toronto General Hospital, and then Sunnybrook Hospital.

And if you look at the reasons for admission, again which are characterized in the large PMA submissions, it is very clear that there is a broad entry criteria, some of which are post-op, and some of which are surgical complications, a lot of which are general ICU population.

DR. NACHAMKIN: I think that this is something that we are going to have to look at more carefully, particularly in relation to the prevalence of disease, and the types of populations, because when you start stratifying these, you are going to start getting particularly small cells.

And so it is unclear whether or not this initial dataset is going to be adequate to address this or not.

DR. WALKER: Well, I think it is a very representative dataset from these ICUs, and it is over a significant period of time, where a thousand patients, for example, were screened initially to go into that.

And of that thousand, 43 percent were suspected of having an infection, and that may be on the day that they come in, or it may be on a day during the course of it.

And most of these patients I think have been -- well, we had no ICU that was strictly purely a medical ICU that didn't take surgical complications, or a surgical ICU that didn't take medical patients.

So the breadth -- in fact, I believe that the breadth of the reasons for admission to the institution or to the ICU in fact adds credibility and robustness to the assay, and it is in fact very generalizable to a very broad population.

Dr. Danner, this is the slide that you asked about and that is the difference between the two groups of patients. So if we dichotomize them based solely on the difference in EAA, essentially this is what the results look like.

And I would take the opportunity to simply also say that this difference between day one and day three, if you get a culture back right away, we would only have day one, all right?

So the reason that we have had to say that is that we have had to recreate when facts occur in a clinical situation, where you take a culture on one day, and you don't get the result back for a period of time.

So this is an observational chart. We have not recreated -- well, we have in fact observed what goes on on a regular basis in the intensive care unit, and it is impossible to do it any other way, and that we get our assay back on day one, just because it takes that length of time.

Culture reports tend not to come back, and in the culture reports, which almost 2,700 cultures were done in this group of patients, the average time for a positive result to come back was three days.

Interestingly, the average result for a negative took longer than that, and I think that is one of the other utilities that we missed in that, and that is that negative cultures, of which over two-thirds of the cultures were negative, take longer to get back, and it takes a longer period of time.

And for that gap, it may in fact be even more significant. But just for the panel to be crystal clear, that we had to pick a moment in time where we would try to make this link between endotoxin in the blood stream, and the presence of GRAM-negative infection in a patient.

And I would also reemphasize that that is the one moment in time where we actually could do this junction. We really can't do it over the course of the seven days, because unless cultures are mandated on every single day, and endotoxin is done every single day, which is not how the study was agreed up.

The issue is that the study -- you know, there were cultures mandated on day one and EAA take on day one. So it is at that point in time, when the moment of suspicion occurs that all these things happen.

The fact that we say day three, you say day three because that is how a clinical practice works, and it is only on day three that you can link the -- when we say day three, that really means when the cultures come back.

And it goes back to Mr. Dawson and essentially what we are saying is on that moment in time there is an opportunity to link the two together. You are linking the culture reports together and you are linking the endotoxin activity together, but you are linking them at that point when the culture result is available.

CHAIRMAN WILSON: Okay. We have time for about three more questions, but before we go on to that, I would like to ask the sponsor that they have shown a number of slides this morning that were not included in the handouts, and we would like to get copies of those for the panel members this afternoon if we could.

In order, it would be Dr. Nolte and Dr. Solomkin, and then Dr. Charache.

DR. NOLTE: Actually, I have a couple of questions, and I think they are quick. One is about the EAA break point of .4, and I have heard several people comment on whether that is the appropriate break point, and I would like to know whether any of the datasets have been analyzed at different break points for positive and negative, and how that impacts the calculations. Is that data available?

MR. DAWSON: Well, the key is what they go into the clinical trial with that is based on some preliminary work up. As I said before, it is always tempting to look for other cut-offs, and statisticians tend to discourage that.

DR. NOLTE: But --

DR. WALKER: Could I answer that? That is a very important question. In the development of this assay, and as I think Marian Heyliger has said, that there was an issue of a .3 being used, and that was when the assay was originally developed by us as a research tool.

And it was re-agents made up every day, et cetera, and we converted this to a manufacturable assay with robust reagent from last year at room temperature. We fought that we should reassess that, because it had clearly changed, and there were changes with respect to certain modifications.

So we ran a pilot trial, and we ran a pilot trial, which was mentioned previously, and it was reported to the FDA. And in that trial, we observed the distribution from our sites that we were going to use, the infections and the threshold, and we then defined that threshold at .4, and we went forward and tested that in the pivotal trial.

So we in no way reshaped the endotoxin activity cut-off level. We set that at .4, and we ran through the trial based on that.

DR. NOLTE: And that evaluation was prior to the clinical trial revealed no value to establishing an equivocal or gray zone for this test?

DR. WALKER: We felt that there were confidence limits at that level that made us happy to go ahead with that as a level. I mean, clearly, as with any other assay, a tuning up and down influences the characters, and the parameters in the assay.

So we believe that the MEDIC trial -- we know that the MEDIC trial was run prospectively with a threshold cutoff of .4, and that would remain in our claim based on the data from the trial, and based on what Mr. Dawson said, really the agreement between that and the culture reports.

DR. NOLTE: And one quick question just so I am clear. The criteria that the CEC used to evaluate these patients was an agreed upon criteria? I mean, is it anywhere in the documentation, or is this 4 or 5 guys getting together and deciding who is infected and who is not?

DR. WALKER: No, it was more formalized than that, and that is that it was based upon -- I mean, there are -- the CEC, as you know, has become a common component of a number of different trials because of the challenges of having information that is in fact clinically useful.

So in this there was -- I mean, we have had -- we have looked very carefully at the definitions that were used, and the people who have been involved in this have been involved in a number of the other CECs.

But what we did was agree upon -- and again drawing from reports that have been in front of the FDA and other areas, what ventilator-assisted pneumonia might look like.

DR. NOLTE: There was one set of standard criteria.

DR. WALKER: Well, no. There was one set -- well, you are absolutely right, and there is one set, and not only that, that was validated. So we sent them out and with the criteria, and we had feedback from that criteria, and then sent out again to have them reevaluate it.

So we had a validation of our protocolized CEC definitions, and then sent out all the data together, and given those specific instructions are given to each individual.

DR. NACHAMKIN: Is this a nosocomial infections definition? I went to their website and printed out CDC definitions of nosocomial infections. Is that the document that you are talking about, in terms of CDC definitions?

DR. WALKER: We have a number of documents.

DR. NACHAMKIN: Because it wasn't referenced in your documents as to which one it was.

MR. DAWSON: Is this 1988?

DR. NACHAMKIN: 19 --

MR. DAWSON: In the title?

DR. NACHAMKIN: This is 1996.

MR. DAWSON: The one that was in the PMA was 1988.

DR. NACHAMKIN: This is by Garner, and it is called, "CDC Definitions of Nosocomial Infections."

MR. DAWSON: It was an earlier one.

DR. NACHAMKIN: This is the earlier one.

MR. DAWSON: This is the one that was in the PMA study and was dated 1988.

DR. NACHAMKIN: 1988.

MR. DAWSON: 1988 was in the title.

CHAIRMAN WILSON: Dr. Solomkin, you are next.

DR. SOLOMKIN: I think somebody made the comment that 80 percent of the patients received empiric antibiotic treatment. I want to know if the 20 percent that didn't, if any of those were false negatives?

DR. WALKER: I think as Dr. Marshall had suggested, there were also -- I mean, the other issue is not just antibiotics, but in fact antibiotics that are appropriate for the organism.

So in a number of the false negatives the antibiotics in fact were inappropriate to the organism that was identified. But your specific question is that if we look at the false negatives --

DR. SOLOMKIN: If you look at patients who did not get antibiotics, or who in other words were clinically considered to have a very low probability of infection, and not warranting empiric treatment, were any of those patients false negatives?

DR. WALKER: I don't know the answer to that question.

DR. MARSHALL: Although I don't have summative data, I can comment that at least one of those patients was a patient with a hemophilus influenza bacteremia, and a wound infection with the same organism, who received no antibiotics over the course of his stay, and was adjudicated a false negative by the assay and survived his ICU stay without complications.

CHAIRMAN WILSON: Okay. And the final question is Dr. Charache's.

DR. CHARACHE: Yes. I've asked if they would put up the slide again, this one. I think it is easy to see when you look at the ones that were defined as false-negatives that there is a species bias, and none of the serratia from these 33 were detected by the tests, and most of the pseudomonas -- well, there were four pseudomonas that were missed.

And there were no E. coli that were missed, et cetera. So there is a species bias on the false negatives.

DR. WALKER: I'm not clear how you have come to that conclusion.

DR. CHARACHE: I have come to the conclusion that when I count the number of serratia ocelots in this particular slide, and there are three, when you list the false negatives, all four seratias there -- and one of those four patients also had pseudomonas in the blood.

So there were no seratias that were true positives according to the definition of the 25 that were true positives. And it is the same rationale for the pseudomonas. There are also no E. coli on the missed ones.

And in the H. flu, there are four H. flus, which is very usual for an intensive care unit. But that is not a bias and that 2 of the 4 were missed, and 2 of the 4 were there.

But the same evenness of distribution is not seen for pseudomonas, or serratia, or E. coli.

DR. WALKER: I am going to check on that. These are the 33 confirmed infections, and out of that there are 8 false negatives. But that is what you have taken your calculation from?

DR. CHARACHE: That's right. I am subtracting the species that were on the false negative table from the ones that are on the total table.

CHAIRMAN WILSON: Okay. Thank you. At this time we would like to move to the open public hearing. No one has contacted the FDA in advance to make comments, but I would like to have ask if there is anyone from the public who would like to come forward and make comments at this time.

(No response.)

CHAIRMAN WILSON: Okay. There being no public comments, then the open public hearing session is now closed. I would like to go ahead and break for lunch now, and I would like to reconvene as close to 1:20 in the afternoon as we can so that we can try and keep on schedule. Thank you.

(Whereupon, at 12:34 p.m, a luncheon recess was taken.)

A-F-T-E-R-N-O-O-N S-E-S-S-I-O-N

(1:36 p.m.)

CHAIRMAN WILSON: Okay. We would like to reconvene the meeting at this time. This is the part of the meeting which is an open committee discussion of the issues that the FDA will present in the form of questions.

This portion of the meeting is open to public observers, but the public observers may not participate except at the request of the Chair. Before we move to the questions, I would like to ask Drs. Solomkin and Danner if they would like to make any comments.

I would like to have Dr. Solomkin go first because he has to leave early.

DR. SOLOMKIN: Thank you. The comments that I have are really in part are primarily confined to the use of neutrophil priming in this disease state.

Priming are in patients like -- or at least in some of the patients that would go into this kind of group, they are reasonably well-defined abnormalities and oxidative function, and priming has not been well studied, but there is some evidence at least in some of these groups that the cells are already primed.

And I am a bit concerned that with these two variables, which is underlying abnormalities in oxidase function from disease; and then secondly the preexisting priming based on either an endotoxin LBP interactions, or other interactions with other substances, such as psydokine, that is -- well, that it would make the likelihood of false negatives very probable.

And the concern that I would have with that is that the patients that -- and they are not basing this on any data that I am aware of, but the patients that I would be particularly concerned about would be the more critically ill patients, where information from this test might really be important and actually affecting their outcome. So I think I would really restrict my comments to that.

CHAIRMAN WILSON: Okay. Thank you. Dr. Danner, do you have any specific comments that you would like to make?

DR. DANNER: Well, I think -- you know, again, everything is riding on the value of a negative test, which for a clinician is a hard issue to wrap your mind around anyway, and to essentially ignore a positive test, because a positive test in regards to diagnosing GRAM-negative infection in this situation is just not good information to base it on.

So when we saw the distribution of the GRAM-negatives shown before, I am concerned that certain types of GRAM-negative infections may be less likely to be picked up by this test than others. I think in vitro testing across a lot of different endotoxins is very different than testing in a person.

And you can find differences in endotoxin in terms of its biological activities just based on how you isolate it, and how much protein is in association with it, and a whole variety of other factors.

So if you are having an outbreak in your ICU with a particular type of organism, and you have been relying on this test, it may be that with events like that, even with whatever you believe this negative predictive value to be, it may change depending on the circumstances and over time.

And I think that is very hard to gage. I also wonder about other sort of interactions with the tests, since the tests do rely on components that are actually in the blood, and I guess with the controls that are done in the three tubes that controls for a lot of that, and with things like complement depletion, and other things that occur during disease, how that might affect the performance of the tests.

And I would like to see this issue of the antibiotics -- you know, if you are basing the negative predictive value on whether your culture is positive or not.

And if you have a population in your ICU where people are largely on antibiotics, and that culture result is a poor gold standard in that setting, and how does the performance of this test change, depending on whether you are looking at a population that has been pre-treated with antibiotics or not treated, or heavily pretreated with antibiotics, like in bone marrow transplant populations and things like that. So, I don't know. That is probably more than what you wanted to hear.

CHAIRMAN WILSON: Okay. Thank you. At this point, I would like the FDA to put up the first question for discussion. Okay. The question reads, "Performance parameters used to describe this assay includes sensitivity, specificity, positive predictive value, and negative predictive value."

And the question is are the diagnostic end-points used in these calculations, CDC criteria and clinical evaluation criteria, appropriate to support these terms, or should alternate descriptive terms be used.

At this point, I would like to open this up for discussion for the panel members. Dr. Nachamkin.

DR. NACHAMKIN: I don't think there has been any compelling evidence presented just with this limited data of the ability of this test to rule out

-- and this has been mentioned before -- is any better than without knowing that information.

The other problem is that even though the test is not indicated, or the response is not indicating that a positive test is going to be used in a diagnostic setting, I am finding it hard as a lab director to figure out how do you separate out -- and if you did this test, the implication of not a negative test.

So if we were to report this out as endotoxin is absent, and use whatever terms that you want, and that's one thing. But if it is present, what do you do? Do you say nothing?

You say that endotoxin is present and we don't know what it means. I think those are dangerous types of things to be reporting out of the laboratory and not knowing how clinicians are going to react.

And I don't think the sponsor has actually done any -- has not addressed those issues in terms of decision making by clinicians in response to these things. They have assumed that everybody is going to take it at the value that the sponsor thinks it should be. But I don't think that is how it would be used in practice.

CHAIRMAN WILSON: Dr. Charache.

DR. CHARACHE: I think two thoughts, and they come back to Dr. Danner's comments. These values that are expressed, predictive values, negative predictive values, sensitivity and specificity, are all stated as predictive of infection, when in fact there is no documentation that it is really predictive of infection.

It is predictive of a positive culture according to certain criterion, in terms of the significance of the positive culture. And because of that it wouldn't help to talk about percent agreement if you are still talking about positive culture, as opposed to something else.

I think also when you talk about using terms such as percent agreement that it becomes very critical that you look carefully on what you want to agree.

If you look at percent agreement on the test as a whole, you have to add all your false positives and whatever to get false negatives. If the goal of the test is to get a no answer, then your percent agreement should be agreement only with the negative test and not lumping the two together.

I mean, if you want a positive answer, you look at the positive side of the column. If your aim is to look at the negative answer, then you look at the negative side of the column.

At the same time we also have to realize that of those that were culture positive, 8 of the 33 were false negatives by the assay. So that also then we have to figure out how to express, in terms of agreement. So that would be agreement on positive cultures.

So you can't just say agreement without defining what it is that you would want to agree as to.

CHAIRMAN WILSON: Okay. Dr. Nolte.

DR. NOLTE: A couple of things. I am still a little confused about the difference between the criteria used, the CDC criteria and the clinical evaluation committee criteria, and how that influenced the outcome of the clinical evaluation, because the CDC did remove a number of potentially GRAM-negative infected patients, and the overall number of infections here is sort of vanishingly small.

So I still am waiting for some clarification on that. I don't think there is any choice but not to use the conventional parameters -- sensitivity, specificity, positive predictive and negative predictive value -- because we have no gold standard here.

So I think that we have to think about these in other terms, and what those other terms are I think is what we have to come to grips with here.

CHAIRMAN WILSON: Any other comments or questions on the first question? Dr. Baron.

DR. BARON: I will just make a quick comment that sort of rides on what Dr. Nachamkin said, which is what does a laboratory do with a positive result. We are struggling constantly in our pharmacy and therapeutics committee about when and how to allow these new anti-endotoxin type therapeutic availabilities to be allowed to be used, and I am very concerned that a positive result in this sort of test, even though it is something like 80 percent of the patients are not infected, would be used as an indicator for anti-endotoxin therapy by a clinician.

CHAIRMAN WILSON: Okay. Any other comments on the first question? Dr. Durack.

DR. DURACK: I am just trying to think logically about this question here, and the key one of course is the one that we have been talking about, negative predictive value.

And I believe that Mr. Dawson made a very clear statement, and if I could read it. "For NPV, determining disease status must be gold standard truth."

So it seems to me that if that is correct, and we don't have a gold standard, then you can't really deal with the NPV. So I see perhaps a choice here.

Either we have to take something like the CDC criteria, and clinical evaluation criteria, and create a quasi-gold standard which would be acceptable -- and I believe that has been done in some other circumstances, but maybe the FDA could correct me and say, well, while we don't have a perfect gold standard, we will have an alternative that is as good as we can get.

And then perhaps be able to talk about NPV, and in the absence of a quasi-gold standard, which is agreed upon by all, I think we have to use alternative terms. And I am just trying to get at the logic of that question. And I doubt that is helpful, but I am trying.

CHAIRMAN WILSON: It does help. Dr. Gutman.

DR. GUTMAN: Well, the question is on here in part to understand your point, and your point is exactly the point of the question, which is that we are trying to seek from the panel a feeling for whether the CDC criteria and the clinical evaluation criteria are strong enough or robust enough, or defined well enough, or clear enough, that we could consider it a tarnished gold standard and support sensitivity and specificity claims.

And even though Pat may not love this, when we don't have truth, then we tend to compare it to a non-truth, and instead of using the term sensitivity and specificity, trying to encourage our sponsors to use percent of agreement, or percent of positive agreement, or percent negative agreement, whenever seems to fit, with the notion that people reading that will understand that it is no more or no less than what it says.

That you are agreeing with something else, whether it is clinical end points, or another imperfect lab test, or whatever you say you are agreeing to.

So it would be helpful to us to know whether the CDC criteria or the CEC criteria from your perspective are close enough to a gold standard that would allow us to cross the line and say that we don't really have a gold standard, but this is good enough, or whether you think it is far enough away that we really should be talking about percent agreement, or whether you have some other option we have not thought of.

CHAIRMAN WILSON: Dr. Baron.

DR. BARON: One of the considerations then, and let's say looking at the CEC criteria, would be to examine the patients who did not have positive cultures to try to figure out if by the CEC criteria, in the absence of a positive culture, that patient would be deemed to be a true infected patient with a GRAM-negative organism.

That would be expensive, and a lot of time and money, but I think that part of the objections to many of the panel members has been on antibiotics or other circumstances that we are missing some patients as well.

CHAIRMAN WILSON: Dr. Charache.

DR. CHARACHE: I just would like to be clear. I don't object to the percent agreement concept. I just want to be very precise on what we are agreeing to, and it seems to me that here we are agreeing that it is not disease or no disease, and it is a positive culture.

I think we do need to know what the significance of the positive culture is along the lines that Ellen has talked about, and also in germs of microbial specificity, and we probably should exclude patients who couldn't have a positive culture because they had been started on antibiotics.

And we don't have any of those parameters and I am not sure that they are available, although they should be in the records of the study protocol that would permit review.

CHAIRMAN WILSON: Other comments? Yes, Dr. Beavis.

DR. BEAVIS: Just the use of -- you asked for our thoughts on the use of the term or the expression of a negative predictive value, and this has been repeated by other panel members, but I think this study highlights two of the difficulties with that expression.

And one is the utility of negative predictive value in a situation like we have here, where it is a low prevalence. And the second is that what you have when you have a high negative predictive value, but it is essentially equivalent to a priori chance of having --

DR. GUTMAN: Well, that's okay, too, but that is a different question. I mean, that comes further along.

DR. BEAVIS: Right. And that is two of the difficulties I think with that in this particular study.

CHAIRMAN WILSON: Dr. Nachamkin.

DR. NACHAMKIN: Yes, but I would suggest that it doesn't matter what you call it, because clinicians are going to interpret it the same way. And if we say that this has a 94 percent agreement with lack of culture positivity, it is going to get interpreted as, or perhaps it may be interpreted as no infraction.

In fact, that is consistently what the sponsor is promoting, that this is a test to rule out infection. And they haven't mentioned that this has anything to do with cultures, per se. The whole document focuses on infection.

And so I am not convinced that changing a term is going to change the outcome of what the result is.

CHAIRMAN WILSON: Additional comments? Dr. Reller.

DR. RELLER: I think we can get bogged down in terminology. We have a test with either of these criteria is incentive in 20 percent of the cases, and including a patient where the utility, if there be any, is an extreme sensitivity to be able to rule something out with a sufficient degree of confidence to take appropriate clinical action, and to not do something, or to do something else based on a reliable negative.

And we have a patient with Klebsiella meningitis who is negative, and I just don't see it. I do not see sufficient confidence in a negative result, and quite apart from all the ambiguities and complexities for the laboratory and the clinician in dealing with a positive result.

But just on the basis on what was proposed and requested, a negative result -- I don't see how we can make it something that it is not. It does not give sufficient confidence to dictate appropriate action.

CHAIRMAN WILSON: Dr. Solomkin.

DR. SOLOMKIN: I think to an extent that I don't really want to directly respond to that, but one of the issues that was raised earlier in regards to looking at the disease, the site of infection breakdowns, to look at these parameters by site may actually provide some information to that.

Because certainly it is very likely, for example, that meningitis would not be associated with high levels of circulating endotoxin; whereas, other infections, perhaps a GRAM-negative pneumonia, may very early on have very high levels of pneumonia.

So it may really be a value to go back -- and as was suggested earlier -- and look at it on a site-by-site basis. And the other issue that I think has to do with most sepsis studies has to do when in the course of the disease you are sampling the patient.

And that really hasn't been controlled for very well with this. I think generally that this was ICU admission, but that is very -- that can vary all over the place, from the emergency room to someone who has been in the hospital for two weeks.

And perhaps taking a look at the data that they have, or perhaps even getting some more data, that we look at and break out those variables that might add to this statistical discussion.

CHAIRMAN WILSON: Okay. Let's move on to the second question then. The second question states that the sponsor stated that the negative predictive value is the key parameter in the assay, and the first part of the question is the NPV of 91 percent adequate and acceptable for this assay.

And the second part is that is the positive predictive value of 15 percent adequate and acceptable for this assay. We are asked to consider the use of a device and how it affects patient management and treatment decisions, and the varying prevalence of GRAM-negative infection in different ICU populations. Comments? Dr. Baron.

DR. BARON: Well, as I had suggested earlier, it seems to me that now that the sponsors have a much larger pool of results in which to evaluate that they could relook at their threshold for positivity, and redo their ROC.

And if they lowered the positive threshold, then they would certainly improve their negative predictive value, and if that is the parameter that they want us to concentrate on, I think that would be one way to go about doing that.

CHAIRMAN WILSON: Dr. Nachamkin.

DR. NACHAMKIN: Again, this is rehashing a lot of things that have been said already, and I guess I am really uncomfortable with the statistical analysis here.

There is such a wide confidence interval on this 94 percent or 91 percent that I am not comfortable that that is in fact what the number is. I think it is going to be much lower and it is going to depend on -- it was mentioned as prevalence and perhaps unit specific.

And this may differ quite from a surgical versus a medical intensive care unit. I think there needs to be a lot more study of this test, and with larger patient numbers to get a better handle on what this range is.

And essentially with 400 patients and 35 infections, I don't think that you can make any judgment as to what the negative predictive value is. And as I mentioned before the positive predictive value is clearly an unacceptable test for predicting infection, and the sponsor doesn't disagree with that.

The question is what do you do with it, and that is a different issue.

CHAIRMAN WILSON: Dr. Charache.

DR. CHARACHE: I would be concerned about a test in this group of patients, which essentially missed 1 of 9, and in fact it is really probably closer to 25 percent of the true culture positive patients.

CHAIRMAN WILSON: Any additional comments? Okay. If we could have the third question. Okay. This question states the primary outcome of the MEDIC study was the documentation of GRAM-negative infection, and the difficulty of determining GRAM-negative infection was shown by the implementation of a clinical evaluation committee to provide a second evaluation of a patient's infection status.

And the question reads should a device performance be evaluated using the CDC criteria, the CEC criteria, or both; and is the use of clinical laboratory information from day one of the study an inappropriate end-point to characterize performance.

I think the first of these questions was largely addressed under the question number one. I think we have discussed that and so let's focus on the second part. And is the use of clinical and laboratory information from day one an appropriate end-point.

DR. DURACK: Just to comment on clarity as we debate this last one. We have to be very careful to distinguish between characterizing performance and characterizing value. I think that is pretty obvious, but we do have to separate the two.

And clinical value and performance may not be the same. I guess we are looking primarily at performance.

CHAIRMAN WILSON: Additional comments? Dr. Baron.

DR. BARON: Well, I understand why they chose to perform the test on day one, and at the same time that cultures were taken, but the data that I would really like to see is how did those patients' test results look on day two and day three, and maybe a combination of those three days, assuming that all these patients are on therapy because they are highly suspected of having a GRAM-negative infection.

And it would be nice to see what happens on therapy. Maybe you could say if your endotoxin comes down dramatically on those three days, then on day three when I am going to make my decision about whether to keep the patient on therapy or not, if the endotoxin stayed the same, then obviously the antibiotic wasn't doing its job.

You know what I mean? There should be more information that would be helpful, as opposed to just the single first day data.

CHAIRMAN WILSON: Other comments? Dr. Nolte.

DR. NOLTE: Well, in reality if this test were to be approved, and in use, the interval of testing would be what? I mean, is this something that is going to be done on admission to ICU? Is it going to be done daily, and depending upon how the patient is doing?

And so having some information -- I mean, clearly, it is going to be used without any guidance from the sponsor, in terms of how it is going to be used. It is going to be used repeatedly in patients I expect.

So having that information I think is an important part of coming to some decision about this test.

CHAIRMAN WILSON: Dr. Sanders.

DR. SANDERS: My comment to that would be that those kinds of things could get hashed out in the package insert. It talks about the clinical utility, and even interpretation of what to do with the positive, versus the negative, and so those are things that could ultimately be fine-tuned. I think there is a bigger picture here.

CHAIRMAN WILSON: Dr. Charache.

DR. CHARACHE: I am also now coming back to the microbiology. We said it missed four of the five of the pseudomonas. No matter if we put it in the package insert, the clinician does not see the package insert.

And they really won't know that if it is a pseudomonas or a serrata, or perhaps some other species, it is not going to have the same activity as it will if it is E. coli.

CHAIRMAN WILSON: Dr. Danner.

DR. DANNER: Two comments. One is this issue of repeated tests. When you have a test which on one draw is positive in I guess about two-thirds of the patients, you worry that if you do repeated tests on the same day or over several days, how many tests do you need to do before everyone has at least one positive test.

And without repeated measures, you really don't know the chances that that might happen. When we have evaluated endotoxemia in our ICU through different technology, we found that endotoxemia as we were measuring it could be quite intermittent.

And I would actually say in terms of people without GRAM negatives, GRAM negative infection being positive for endotoxin, even though our technology was very different than what was used here, the results and the confusing picture that emerges from trying to measure endotoxin in the blood is really not that different.

You know, the data and sort of that confusion in the people that are positive even though they have a Staph aureus infection and things like that, have been part of this literature for a very long time.

So I think that is a concern. And in terms of what performance criteria, I think the problem that you can't work out in the package insert is that there may not be a performance criteria that makes any sense for this.

If you take people coming into the ICU who -- the physicians taking care of them, the intensivists and infectious diseases attending seeing them, who give them a diagnosis of septic shock, say that this person came in clinically, and I believe that this person has septic shock.

And the literature suggests that you can only culture what you think is the causative organism, maybe 50 percent of the time. Sometimes in some studies less.

So the criteria that all of this has been based on, the positive culture as being indicative of infection, there is a group of patients who clinically are believed by the physicians taking care of them to be infected, and to have a very severe manifestation of infection, but yet not have a recoverable organism by that criteria.

So what that means in terms of the performance of a test like this, I don't know. I don't know how you could really accurately gage performance.

CHAIRMAN WILSON: Other comments? Dr. Sanders.

DR. SANDERS: I just want to go back to the issue of the package insert, and by no means was I meaning that the clinician would actually read the package insert and base his or her clinical judgment on that.

That would be used as a guide for the laboratory personnel and the laboratory director to then aid the clinician with the ultimate interpretation.

CHAIRMAN WILSON: Okay. Any additional comments on number three? Dr. Nachamkin.

DR. NACHAMKIN: This wasn't presented, but I think in some of the study documents you asked if the -- and this is directed to the sponsor. But that you asked the physicians their pre-test estimate of infection, and you had some kind of scale if I am not mistaken.

Did you actually look to see how well physicians just predicted the absence of infection based on your interviews with them?

CHAIRMAN WILSON: Dr. Walker, would you like to respond?

DR. WALKER: This may be an example of the same issue that the panel is grappling with, and that is the challenge that we are faced with in these patients in the intensive care unit as to what is real and what isn't real, and what we can know about a patient.

So that was our proposal as well for the same reason you had thought. Our challenge in that was compliance amongst the physicians. So, in fact they did not fill that form out adequately enough for us to make significant.

And it really goes back to the question of this issue is the patient infected or not infected, and that essentially became the question. And in fact the issue was not even site specific.

We don't have information that was useful to interpret that. We infer that they -- I mean, they put the patients into the study based on a decision that the patient was and they would act upon that.

But I think that what is being highlighted by the panel discussion is two things, and that is that the suspicion is high and the reality is low, and there is a big gray area in between.

CHAIRMAN WILSON: Thank you. Any additional comments? If not, let's put on the fourth question. The fourth question states did the endotoxin assay meet the primary objective of the MEDIC study; that is, to exclude the diagnosis of GRAM-negative infection in critically ill patients admitted to the ICU of suspected infection.

And we are asked to consider the bioavailability of endotoxin in the setting of GRAM-negative sepsis, and some organisms shed more endotoxin than others.

And the issue of the binding of proteins to lipopolysaccharide, and clearance of endotoxin from the circulation; and finally the limitations in the devices ability to detect endotoxin from non-hematogenous infection sites early in the course of infection. Comments? Dr. Baron.

DR. BARON: There were 10 false-negative patients, and 11 false-negative sites in 10 false- negative patients, and I am just looking back, but there was something like 33, and so it doesn't look good.

CHAIRMAN WILSON: Other comments? Okay. If there are none, then let's have the fifth question. The question asks what recommendations and suggestions should be provided to improve the labeling for this assay. Does anyone have any suggestions for that? Dr. Nachamkin.

You can cut me off if this is not related to that question. It has to do with the specification that a certain tube be used for the assay. You specifically said that the EDTA tube in a given catalog number had to be used for this assay.

Did you look at other suppliers and it was just called a sterile tube. Don't these things have to be certified as endotoxin free, and is that product the only one that is endotoxin free? And has it been tested, and did you test other suppliers of EDTA containing tubes?

DR. WALKER: That's a good question, and I am going to ask Dr. Romaschin to answer that question. The evolution of this -- I mean, we did start with those extraordinarily expensive certified endotoxin free tubes.

But for a number of the reasons that Dr. Romaschin mentioned, our ability to use generalizable tubes is now confirmed.

DR. ROMASCHIN: Yes, we chose EDTA tubes for two reasons. Number 1, the previous studies by Robert Allen, who is one of the pioneers of neutrophil chemiluminescence suggested that in order to preserve compliment activity over reasonable periods of time, that was the optimal tube.

Secondly, all the BD lot numbers that we have ever tested have tested negative for endotoxin by LAL assay. We have not tested other suppliers, but certainly all the sources of EDTA tubes that we have tested have been negative. That is the only comment that I can make.

DR. NACHAMKIN: So that implies that in your proposed labeling that you would have to specify only that a BT tube could be used currently?

DR. ROMASCHIN: Yes, unless we tested other ones.

DR. SOLOMKIN: But the implication is that that is really saying they are endotoxin free because he said they have tested all of them. So I would think that the language would have to be using tubes that have been shown to be LAL negative.

CHAIRMAN WILSON: Dr. Durack.

DR. DURACK: Just a specific point about labeling. I would suggest that adding to the proposed labeling, the point about antibiotics, which has been studied, has interfering substances and the only drug mentioned at the moment is steroids, and specifically mentioned in the proposing labeling.

I think there should be other common drugs, such as aspirin and common cardioactive drugs, which could well be added to the list of interfering substances that do not interfere.

CHAIRMAN WILSON: Dr. Baron.

DR. BARON: It looks like the test performs better for sepsis in blood, as opposed to like pneumonia. So maybe the labeling could be a little bit more specific about the type of infectious disease that the negative test really feels comfortable ruling out.

CHAIRMAN WILSON: Dr. Nachamkin.

DR. NACHAMKIN: I would just disagree with Ellen, because I don't think there is enough numbers for any particular type of infection to say that you can rule out any of those.

There is some suggestions, but there is only how many cases of bacteremia or there is very few. So I would not base a specific label on those small numbers.

DR. BARON: Yes, I didn't mean exactly just to go for it from this point, but that that would be a potential way to circumvent some of the problems that we have discussed if the sponsor went back and relooked at their data, and came up with other suggested labeling requirements.

CHAIRMAN WILSON: Any further comments? Dr. Reller.

DR. RELLER: I think one should defer the labeling on how to use a product until one has a product to use.

CHAIRMAN WILSON: Any further questions? At this point, I would like to ask the FDA if their questions have been addressed completely, or if they have any other points that they would like us to address?

DR. GUTMAN: No, you have give us plenty. Thank you.

CHAIRMAN WILSON: Okay. We are a little bit ahead of schedule now as we have caught up, and so at this point I would like to move to the open public hearing, and if anyone would like to make a comment, if they would please come forth. If not, then we will close the open public hearing.

At this point then, let's move on to the sponsor's response, and if the sponsor has any additional comments that they would like to make before the panel at this time.

DR. WALKER: Thank you very much for this opportunity to respond to some of the questions that were raised. We would like to take them essentially in the order that they were presented this morning, followed by some of the discussion that has gone on in the panel discussion today.

I will address the first one and that is the CDC criteria, which were based on the CDC website, and while they are based predominantly on the article that was referenced in the PMA, because at the time of the creation of the protocol that in fact was the article that was available.

Clearly, we stay up to date with both CDC and FDA, et cetera. On the other hand, we would clearly accept that it is reasonable to look at that and look at our criteria, which are in part of the PMA to determine if there are any differences whatsoever between those two.

With respect to the role of the CDC, I thought we should probably have one more discussion on that, and then I am going to ask Phil Dellinger just to make a comment on this, because it is clear that we have been struggling with the two issues.

One is the application of statistics, and secondly the evaluation of end-points, for a very long time in the intensive care unit. And particularly struggling with them with this assay development and conducting this trial over the last 5 or 10 years.

So I think it is important that we have a little bit more discussion on this issue of in fact the role of the CDC.

DR. DELLINGER: As a point that I think has been made multiple times by both panel members, as well as people here from the sponsor, is perhaps that I think that all of us would agree, or I hope, that there is no gold standard single test to be able to say someone does or does not have GRAM-negative infection.

When we started doing large multi-center clinical research trials throughout the world, it was very important to try to get as close a gold standard as we could get for who was actually infected.

And unfortunately we were unhappy with any type of template that could be applied in a purely objective fashion, as far as the data that was on a template, and to say that if they had one of three, or two of four.

The performance was just not good. It was good, but it was not to the level that we wanted in clinical trials. The clinical evaluation committee was developed and actually studied in a prospective scientific manner to see if a group of experts, not using any pre-designated criteria, but capable of using any criteria that they wanted to us -- the CDC criteria, culture positive, white count, whatever -- could sit as a group of experts, and with a pre-defined system of adjudication if there was disagreement, could decide whether someone was or was not infected.

This has been shown to produce the best performance to this date for predicting who has infection, or at least let me say the community considers that as currently the best way to say that someone does or does not have an infection.

But it really doesn't use any pre-defined criteria that could be presented to this group. But I think there is consensus that this group of experts does provide the best predictability of infection, and that is what was used in this case, the CEC, and that was the approach that they took.

DR. WALKER: A question was asked earlier today about the effect of precision on the assay, and I think we should look at that from two different aspects. The first is the actual precision that was recorded in these 10 centers around the world.

And then the statistical impact of that, and so I am going to first ask -- could we have Slide 36, David. And, Alex, would you speak about the precision of the assay?

DR. ROMASCHIN: So, yes, there were two points that were brought up, and the first point was what is the overall precision of the assay, and in effect we calculated this on a weighted basis from all of the clinical trial sites.

We just drew the precision and weighted it by the number of patients enrolled at the site, and it turned out to be 11 percent CV, which is the range that you would expect for a manual immunoassay type of procedure.

And Andy can comment on what effect this would have on the estimation of the negative predictive value.

DR. WILLAN: Well, negative imprecision is one of the reasons why the test properties aren't perfect. I mean, it is one of the reasons why the NPV is not one, and it is one of the reasons why the sensitivity is one.

So that is how the effect of this imprecision affects the statistics. I think a question was asked and I missed it early on.

DR. WALKER: So essentially the statistical -- the results that we have put forward with NPV confidence limits takes into account all the precision challenges that are apparent in the assay. So that this is not an addition, but rather this is factored into all of the statistics that we are presenting, because these are the statistics with a precision of 11 percent or a CV of 11 percent.

We had a discussion about false negatives on a number of occasions today, and I think it is important that we address those, because unfortunately because of the way that these numbers have been presented in our struggle in order to present the reality of the situation in the clinical intensive care unit, that we have used both CDC and CEC, and there is a difference in that.

So, Dr. Baron, I'm sorry, but the numbers that you have quoted, in fact you took the worst number from one side, and the best number from the other. It is not exactly a fair comparison.

But there are some very specific important issues about the false negatives that I think we need to address, and I am going to ask Dr. Marshall to begin the discussion on the false negatives.

And this has to do with two aspects of this, both the allocation of them, and as well we will have Dr. Romaschin talk about the ability of the endotoxin assay to pick up different endotoxin strains.

DR. MARSHALL: Okay. Thank you very much, Dr. Walker. First of all, the template that we used for this clinical evaluation plan was actually derived from a study that we published about 3 years ago in the New England Journal of Medicine, looking at two different strategies for stress ulcer prophylaxis.

And those data show very clearly that depending on the definitions used that the prevalence of the disease varies quite strikingly. We used that particular model and saw very similar kinds of results.

And for the reasons that Dr. Dellinger has outlined, opted to take the clinical evaluation committee as the best available estimate of true prevalence.

And I have to say that as somebody who works in an intensive care unit the reality is that we ignore information all the time; a positive triplopia, a positive Fletcher. A high-elevated blood sugar doesn't mean diabetes, and a positive culture doesn't mean infection.

And if we are only -- you know, if we are sensitive that it is 80 percent, it really begs the question how do you determine sensitivity in the absence of a gold standard.

What I would like to speak to about though is the issue of the two organisms that were raised as potentially missed by the assay. One was Serratia, and we have gone back and reviewed the numbers.

There are 3 of 11 missed patients who had Serratia infections, and 2 of 43 patients who weren't -- I'm sorry, two of -- well, yes, 2 of 43 in the CDC criteria had Serratia.

With pseudomonas, it was 5 of 11, versus 10 of 43. The numbers are very small. They are not large enough that we do a Chi score on them, and come up with statistically significant results.

I think this speaks to two questions. One is are we actually measuring endotoxin, and I believe that the data that Dr. Romaschin has shown shows that they were both highly sensitive and highly specific for endotoxin.

The second question is when we detect it in conjunction with an organism does this mean an infection, and these in fact are two organisms that typically show up late in critically ill patients, and whose pathogenicity is uncertain.

So it is equally plausible that these were not infections, as it is that they were missed infections. I would like to address one other issue, and that was I think a very important one that was raised, and that is about Klebsiella meningitis.

This was not a patient who came into the emergency department fomically septic and proved to have Klebsiella meningitis. This is a patient who had been in the ICU and had an intracranial screw in place, and cultures from an intraventricular device yielded the Klebsiella. So it may have been a device-related infection as with that particular aspect.

DR. WALKER: Alex.

DR. WILLAN: I just wanted to address this issue again of sensitivity, and with regard to these two organisms. Serratia marcessions, whether the LPS is presented in pure form or whether you grow the bacteria and then sonicate them, or extract them, and put the material in the circulation, the serratia is the most sensitive LPS that the assay detects.

So the fact that it was missed isn't -- I do not believe because the endotoxin was not in the circulation. The question is whether it has shed or not.

But certainly that is one of the best organisms that we can detect, and similarly pseudomonades are very easy to detect in this assay. So I don't think that these are issues of analytical sensitivity.

DR. WALKER: Alex, while you are there, Dr. Solomkin asked a question about neutrophil priming in these patients, and I think it speaks to the veracity of the assay in this entire patient population.

And outside of this particular use, and in effect leading up to the discovery of this particular assay was a great deal of work by Dr. Romaschin in neurobiology. So I think it would be appropriate for him to make a few comments on your comment.

DR. WILLAN: We were equally concerned about the priming effects, particularly in people who already have multiple priming influences, but we are incredibly surprised at most neutrophils, even ones that have been banged around by cytokines -- in fact, in our assay there is a built in control for this.

And this is a plan that I want to stress, that we challenge the assay with pre-formed immune complexes, and set that as a maximal signal. So if the capacity to be primed is lost. We don't get a signal.

And surprisingly that occurs in a very, very small percentage of patients, less than 1-1/2 percent of all the patients we have studied. So despite the fact that many of these patients have actively activated neutrophils, neutrophils have a 200 to 300 full capacity to be up-regulated.

And many of these patients never ever reach that capacity, and we control for that as part of the assay. And when that capacity, we call it a non-assay.

DR. WALKER: Okay. Thank you. Dr. Marshall, one of the other questions that was brought up really by Dr. Danner this morning was what is the distribution, and what is the relationship of endotoxin activity in patients with GRAM-positive infections, and as you recall, we only have one patient with a confirmed fungal infection, but a significant number of GRAM-positive infections.

DR. MARSHALL: Thank you very much, Dr. Walker. First of all, I do have data that were asked for about the number of patients whose false-negative GRAM-negative infections were not on antibiotics, and that was 4 of 11 that were not on antibiotics at the time.

The levels of GRAM-positive, we have some data, and these have just been calculated for me now. The mean level of endotoxin activity in patients with GRAM-positive infection, the end was 46, was .56. So clearly we were detecting endotoxin in patients who had GRAM-positive infections.

And in fact the likelihood ratio data that we have for GRAM-negative infections, the likelihood ratio .71, and for GRAM-negative, a .56; and for GRAM-positive infection, meaning if you had either GRAM-positive or GRAM-negative infection, you were more likely if you were endotoxin, you were less likely to have either of those than otherwise.

But our claim is not -- this kind of becomes counter-intuitive when the claim is not being directed towards the possibility that endotoxin can make a diagnosis of GRAM-positive infection, although the comment has been made that infection may increase the availability of endotoxin from the gastrointestinal tract. So I think those are the data for GRAM-positive.

DR. WALKER: I would just like to direct to other questions, or two other responses to Dr. Danner, and it has to do with the actual pathophysiology of endotoxin, because this truly is a fascinating area.

And while we don't want to -- we are not allowed to wander off into areas of discussion of endotoxin as an entity in itself, we are focused here on the relationship between endotoxin and infection, which is our claim in front of the FDA, which brought up a couple of points.

And that is that in your work, with which we are quite familiar, the issue of intermittent release of endotoxin, and clearly we have been concerned about that.

But we did a great deal of clinical studies and pre-clinical studies looking at this through a number of patients for a long period of time.

And we did not find the fluctuation of endotoxin on a regular or hourly basis. If we had seen that, clearly I think we would have redone the protocol.

We have put a great deal of weight on one assay. In other words, one daily assay, with the idea that a useful assay wouldn't be perhaps having to be repeated three or four times in a day.

So that has been our findings, and I also would have to say -- and I am sure that you will have some comments on this, but in our -- I mean, we began working with endotoxin using the ALA assay.

And with all due respect that has not been FDA approved because it has not proven to be useful in the clinical setting. Now, we also found that it was not useful in a clinical setting.

And in our pre-clinical studies of a great number of septic patients, what we found was that the LAL level was actually the lowest in the patients that were the sickest, with most likely to be septic, and in fact in our studies with the highest level of endotoxin level.

We also found that the LAL assay as you well know is not something that you can do on a regular basis. It has to be batched, because you have to develop a standard curve. So it is not actually in the same category as ours, which is a test that can be run within a short period of time.

And as Dr. Romaschin said, one at a time, or in a batch if necessary. But we found a great deal of variation in repeating the same samples using the LAL assay.

We found that conditions could change very little and find a great deal of difference in the actual level that was reported by the LAL assay. So that has been our experience with that.

It has not been our experience with our own assay, because we repeated numbers in the PMA, and that is that within run, between run, precision, et cetera.

So that may not be a complete explanation, but it is more information in the area of the pathobiology of endotoxin release.

DR. DANNER: I would like to on record to say that I am in no way advocating the LAL assay. As someone who has used it for research purposes, I agree with all the comments that you made about it, and the difficulties with using that test.

And clearly the real advantage of your test is the fact that it can be done so quickly, and not require the standard curve and the other preparation, and the things that one has to do to handle false activity, the suppression of activity, and all the other problems attendant with the limulus lysate assay.

I guess ultimately though this comes down to even though your test is a faster test, and even if we assume that it is always measuring endotoxin when it goes above .4, which are still to my mind assumptions. I don't know all of the possible conditions that might occur in clinical blood that may make that not true.

The question is whether this is really useful clinically, as opposed to being useful as a research tool, as a research tool to -- well, for instance, make some determination for investigational agents directed against endotoxin, or as somehow investigations into bacterial products and sepsis.

And the issue that Barth brings up I think is the real question, is would -- if you did a study where you did your test, and you gave the result to half of the clinicians, and to half the clinicians you didn't give the test, would the patient be better off or worse off with that information, and I don't know the answer to that at this point today.

DR. WALKER: It sounds suspiciously like a post-marketing study, and I couldn't agree with you more that that would be interesting. I think you have to understand that we have been hampered in our ability to understand a lot of the biology that goes on because we have not had a reliable test for endotoxin.

We are not standing in front of you today to say that we have a test that is going to unravel all of the intricacies and the unusual aspects of endotoxin.

We are saying that we have an ability to measure endotoxin, and it has usefulness in a clinical situation.

DR. DANNER: But your test for endotoxin has a lot of the same -- you know, which may be just part of the biology in it, but it has the same problem, where people with GRAM positives have positive endotoxins almost as much as people with GRAM-negatives, which was a problem seen with the old limulus lysate assay.

And that's really where I was making a comparison between the two tests. In other words, it is not like you have a test that is only positive when you have GRAM-negatives, a GRAM-negative infection.

And again that could be related to the underlying issue of what puts endotoxin in the blood. And Ron Elin, who worked in this area a long time ago, back in the '70s when he was at NIH, and then later I worked with him on some of the studies that I did, pointed out that the amount of endotoxin in one GRAM-negative bacteria is so small, in the phemtogram (phonetic) range, that you would need more bacteria in the blood than you normally get in order to detect the positive test.

So even in the setting of a GRAM-negative infection, the endotoxin isn't just because you have bacteria in the blood. It is from shedding and coming from other sites, or maybe crossing the -- well, you know, we don't even know that if you get GRAM-negatives out of the lung that you have pseudomonas pneumoniae, is the endotoxin that is circulating from that pseudomonas, or is it just from other bacteria in the blood, and it is not even pseudomonas endotoxin.

I don't think anyone can answer that question.

DR. WALKER: You have actually restated our situation in a particularly positive way and I am grateful for that, because you have actually brought up the points.

We don't disagree with anything that you have said the bottom line is, because our claim is not what the presence means. Our claim is what the absence means, because you have very accurately said that there are many potential causes.

We would like to unravel those. I do believe that we have shown enough evidence that shows that we are very specific in our ability to pick up endotoxin.

We have not found anything that interferes with this assay that causes a positive response in the situation that you have described; neither a drug or another form of organism.

So we have not found one that has done that. And the issue of where it is coming from is a very good question, and I can't answer that. And I would go back to the question that Dr. Baron said, saying you don't want this to be used to treat anti- endotoxin, or at least an indication for anti-endotoxin therapy.

First of all, there is no FDA approved endotoxin, anti-endotoxin therapy. It doesn't exist. It would be nice if it did, and it would probably save some lives, but so far it doesn't.

And the issue is that we don't -- you know, we are not making claims on that. Endotoxin is a peculiar individual, and up until now it has not -- it has alluded any successful measuring device. We believe that we have a successful measuring device.

So the actual intricacies and the contributions to illness and endotoxin may have to be -- and to be honest with you, are yet unknown.

DR. DANNER: Getting back though to your negative predictive value, you still have a problem there because you didn't miss a clinically significant number of people who did have GRAM-negative infection.

So basing clinical decisions on that test and saying that this is less likely, well, it may be a little less likely, but there still was a significant proportion, a clinically significant or relevant proportion, that were negative, but had infection.

And then the other thing is that all of those numbers are still based on the tarnished gold standard of a positive culture in a population that I am sure was heavily pretreated with antibiotics prior to some of those even initial cultures.

And so if you take the other side, then I would say that there were infected people in your population that just didn't have a positive culture. So your negative predictive value would even be lower than what you are currently estimating it at.

DR. WALKER: Well, we don't know that. We never used to think that.

DR. DANNER: Well, as a clinician, I believe that is absolutely true, and if somebody cane to me with your test, I wouldn't change everything that I am doing based on the information I heard today.

DR. WALKER: I couldn't agree with you more, and actually I would even go further and say I wouldn't change anything that I was doing.

DR. DANNER: That I disagree with.

DR. WALKER: I just wanted to make a comment on that, because I think we have had a -- I think it has been a very good discussion about the tarnished gold standard of the diagnosis of infection. If somebody has a better one, I would be happy to put our test up against it.

We are challenged, and we have to deal with what we have, as does the panel members, and as does the FDA. There is not a perfect assay. And if there was a perfect assay, I think we would have a much easier -- I'm sorry, a perfect diagnosis for infection.

And I am sure that we would have a much easier course. If there was another course for measuring endotoxin, we would have a much easier course. This molecule is difficult to measure as we have talked about.

Now, I really want to reiterate that we are not saying that you are going to change your view of the patient management based on this test. I would hope to think that you don't base very much patient management based on one test.

In these complicated patients, we very seldom make a decision based on one test, unless that one test --

DR. DANNER: If this test isn't changing my management, then why am I buying it for the patient? Why am I ordering it, looking at it, and charging the patient for it?

DR. WALKER: There is a couple of questions in there. I mean, I think we have talked about what information this assay may offer early on, and while Dr. Reller has said this is a non-test from a statistical standpoint, I would beg to differ.

And that's because clinical judgment has resulted ina 92 percent false positive rate. As we understand the actual incidence of a truly confirmed infection is low and that's the case. That's the truth.

We have to deal with the facts as they come. On the other hand that is not how clinicians behave. And so far there is nothing that the clinicians can believe in that gives them any comfort at all that that patient isn't infected.

Whereas, only 8 percent of them are. So the issue is how do you find that vast majority of patients who aren't infected. So the ability of our test at that point is to convert that 92 percent of false positives clinically and reduce that to 128 patients out of 128 were true negatives, and their course then would be altered by virtue of the fact that those patients are unlikely to have an infection.

And how it is going to be altered depends upon the algorithm and decision making, and the entire clinical situation to pick out the patient. And certainly in a patient with fulminant and GRAM-negative infection, we don't need to test to rule that out.

DR. DANNER: Again though you are going back to the -- you are saying a hundred percent of these were suspected of being infected, and only -- and 92 percent weren't infected.

You can't say that. That is absolutely not supported by the literature of what patients are like in the ICU, and what people think clinically. And if you go and do your tests, and do the culture, and stop antibiotics on those other 92 percent, that is the wrong management of those people.

DR. WALKER: I understand what you are saying, and I think that has been discussed in the application of the NPD, and to be honest with you, I am not disagreeing with you.

I think we have information to add, and it may not be best expressed as a negative predictive ruling out in its entirety GRAM-negative infection, and I think the proposal put forward with respect to agreement is something that clinicians can relate to.

And as a non-statistically bent clinician, the concept of a negative predictive value is not particularly different with respect to essentially an agreement with a clinical situation. So I understand what you are saying about that, and I think we should reflect that.

CHAIRMAN WILSON: Dr. Janosky.

DR. JANOSKY: Dr. Walker, there must have been an oversight, because I didn't hear an answer to the question that I had asked this morning.

DR. WALKER: Your question on prevalence?

DR. JANOSKY: Yes.

DR. WALKER: Well, there would be two answers to that question. And that is that if they don't want to use the negative predictive value to evaluate this patient, then in fact the prevalence becomes difficult to evaluate.

But we can give you -- we actually provided the NPV from different sites, and also the prevalence of the infection from -- well, we have that.

DR. JANOSKY: Good. That was the information that I was looking for.

DR. WALKER: Yes.

DR. JANOSKY: Good. Can I just make a comment to an earlier discussion while we are waiting for that?

CHAIRMAN WILSON: Please.

DR. JANOSKY: I feel a little uncomfortable with the way that the word agreement is being used, as sort of a catch-all, and that we can't do these statistical criteria, and so let's use agreement in sort of a lesser way.

Well, agreement in and of itself also has methodology appropriated with it, and if you just use it as a catch all because we can't do the other. You are placing all that methodology and saying it doesn't exist, where in fact it does exist.

So the term agreement in the way that it is being bantered around here is actually being used in correctly. So I would caution us in thinking that that is the way to deal with this issue of not using NPVs, sensitivity, specificity, and keeping track of all the methodology that does go with the assessment of the agreement.

It is something that we can come back to later, but it is just an issue; and if I could see those numbers. Do you have them?

DR. WALKER: As soon as the computer warms up.

DR. SOLOMKIN: Let me just ask you one quick question, Dr. Walker. I may have missed this and so I apologize, but what is the sense of a positive test in a normal population, ambulatory, and no reason to suspect disease?

DR. WALKER: In the instance of a positive test in a normal population walking around, it approaches zero. It is about one percent. It is interesting though that we have done this in smaller studies, looking at the incidence of endotoxemic in other areas.

And it is interesting in that the incidence is far greater than that in certain patients. And we certainly see things that increase the level of endotoxin in an ambulatory patient, an interesting one of which is cigarette smoking, and it is an interesting observation in our cardiac patients.

DR. NACHAMKIN: While we are waiting, I just have a technical question. In looking at the analytical specificity studies, I noticed that for the GRAM-positive organisms that were tested, you mentioned in a document that it was a pulled extract of a variety of different positive organisms, and

they actually weren't tested individually.

So it is not clear to me that that is a reflection that in fact is specific enough. Secondly, you use serratia as a source of antigen to test the specificity for fungal pathogens. Why didn't you use real pathogens, like candida, cryptococcus, et cetera, for those studies?

DR. WALKER: We have done further studies in both of those areas, and I will ask Dr. Romaschin to more fully elucidate those.

DR. NACHAMKIN: And one last thing. Do you have any evidence that if you mix GRAM-positive organisms with GRAM-negative organisms that you can mask the reactivity of the GRAM-negative organisms in your assay?

DR. ROMASCHIN: I can comment on the fact that we have tried heat-killed in live aspergillus and candida albicans, and they don't give a response. With regard to the GRAM-positives, we have also tested them individually, and not as a mixture.

We have tested each of those bacteria individually, and if we use mixtures of bacteria we have actually not done those studies where we have used GRAM-positive and GRAM-negative added mixtures.

DR. NACHAMKIN: So once again maybe it is minor in context of everything else, but is it possible that the patients that were actually missed had some other GRAM positive organisms, whether they be colonized or infected, that could have masked the reactivity in those patients?

DR. WALKER: In the clinical situation obviously polymicrobial infections are not uncommon, but our assay has not been disadvantaged by that particular.

So that we actually have -- and if you look at the distribution of the GRAM-positive organisms, 38 of those had an endotoxin activity greater than .4, and 10 had less than .4, which is essentially the split that we would normally see within this patient population.

So I don't believe that there is any reason to think that there should be any interaction between the GRAM-negative, probably LTA or something like that.

DR. NACHAMKIN: But you haven't looked at that specifically?

DR. WALKER: Well, I can't say we have not looked at it completely. We are in the process obviously of further developing a GRAM-positive assay, looking specifically at a typical or suitable antigen, like LTAs.

So we have clear studies done on that, and the actual mixing of LPS and LTA I think we have not done. But we have in the clinical situation, in the vivo situation, we have had situations where there have been polymicrobial infections, and we have not found those to be consistently in one category or the other with respect to known diagnosis.

DR. WALKER: Were you able to see the prevalence?

DR. JANOSKY: No, there is nothing up there.

(Brief Pause.)

DR. JANOSKY: So there are two sites that had approximately a hundred patients in each, or excuse me, the three sites. Which ones are those?

DR. WALKER: Site Number 1 is Toronto General, and Site 5 is Brussels, and Site 10 is Sunnybrook.

DR. JANOSKY: Okay. So it is 11 percent, 6 percent, and 7 percent? Is that correct?

DR. WALKER: Yes.

DR. JANOSKY: Based on -- is that the CDC and CEC?

DR. WALKER: They are both up there.

DR. JANOSKY: Okay. And CDC is on the right. Okay. So based on CDC, the numbers are quite different; and based on CEC, the numbers are quite different across sites; and those are prevalence values, correct?

DR. WALKER: Yes.

DR. JANOSKY: Do you have the same things for your negative predictive values?

DR. WALKER: That was supplied to the FDA, which was an NPV on a site-by-site basis.

DR. JANOSKY: Do you have that where you could tell us those numbers? I know that I had looked at it at some point.

DR. WALKER: I'm wrong. I take that statement back again. Obviously, it would be difficult to have an NPV on a number of those sites where in fact the incidence of GRAM-negative infection was so small.

DR. JANOSKY: So your prevalence values are different, quite high actually?

DR. WALKER: Yes.

DR. JANOSKY: And your NPVs are not?

DR. WILLAN: I doubt very much that it would be another one by chance wouldn't you say?

DR. JANOSKY: What are you referring to when you say that? I'm sorry.

DR. WILLAN: Well, I am looking at the three sites where there is more than a hundred patients; 11, versus 6, versus 7. I don't think that is statistically significant.

DR. JANOSKY: Well, your ends are so small, and so you are probably not going to pick it up.

DR. WILLAN: Well, they are over a hundred and they are not that small.

DR. JANOSKY: That would be considered small if you are looking at different and in low proportions like --

DR. WILLAN: Yes but the fact is that they are not statistically significant. You can't draw a conclusion that they are different. You either say you don't have the evidence or you conclude that they aren't different.

DR. JANOSKY: If you are not giving me the NPVs, I can't really tell what impact it has.

DR. WILLAN: Right.

DR. JANOSKY: And you are telling me that you don't have them available to us right here; is that correct?

DR. WILLAN: I am just saying that I don't think that you have reason there to believe that they are different between sites based on that evidence.

DR. JANOSKY: We have reason to believe they are different; maybe not statistically different.

DR. WILLAN: I don't think that those two statements are different. I think you are contradicting yourself.

DR. JANOSKY: Well, as we both know being biostatisticians, there is a difference between saying something is different and saying something is statistically different.

And those numbers are different. They might not be statistically different at different points, but that is a statistical argument.

DR. WILLAN: They will never be exactly the same would they? They would never be exactly the same.

DR. JANOSKY: By chance, they could be or they could not be, but that's a statistical and theoretical argument that perhaps shouldn't have an argument. Let me ask --

DR. WALKER: Let me say that there was a rigorous examination of the characteristics of the patients at each site, and I think you are familiar that with the trials in the critical care setting is often having to use multiple centers, and to pool the data in order to have meaningful results.

But in each of these sites, all the characteristics, all the demographics, have been looked at very carefully, and provided to the FDA, and reviewed, so that the pooled data would appear to be appropriate.

DR. JANOSKY: Okay. Let me ask one final question in terms of some of this issue. What if I would postulate that the actual sample size for this particular study was a hundred or slightly over a hundred, 125?

So you are actually basing your outcome on this particular study on about 125 patients, because you are using a negative -- you are saying that less than a .4 actually is an inclusionary criteria for the study, because you are not taking a look at anybody who has greater than .4.

You are saying discount all of those data for anybody who has test value greater than .4, and we only want to pay attention to those that have a test value of less than .4, because that is your conclusion, that it is only based on that particular group.

So if that is so, then I would postulate that the sample size that you are using for this particular study is slightly over a hundred. It is about 125.

DR. WALKER: The same size calculations were reviewed with the FDA for all of the reasons that you have suggested, and the sample size was set upon identifying a number of patients with a negative -- I mean, we have to have a large enough net to find an appropriate sample size of patients who we predicted would have a low endotoxin activity.

Obviously, we didn't know that, and in our pilot studies and in our pre-clinical studies, it appeared to be about a third of the patients. So in order to make meaningful statistics on the agreed upon sample size was that we needed to have about a third of our patients to fall into that category, which is essentially what they did.

And so basically what you are saying is that out of the 408 patients, 128 of them had negative values.

DR. JANOSKY: Right. I am not questioning the sample size estimation a priori. I did take a look at that and I am not questioning that. I am just questioning the number that you used to say that were actually studied, because the results are only based on that negative group.

The results that you are talking about, in terms of let's pay attention to the negative predictive value, if that is what we are going to call it, is only based on slightly over a hundred, and you are telling us to discount all the others because you don't want those to play into our decision, and so you are saying don't pay attention to the negative predictive value and all those other groupings.

DR. WALKER: From both a statistical standpoint and from a pathobiology standpoint, we are saying that we simply cannot attach significance with respect to infection to a level above .4, and that is the question that we were essentially asked to prove by the FDA.

Is there an association between a negative value and the absence of infection. So what you are saying is right, and I am not arguing the numbers. The numbers are the numbers as they are.

But to say that we didn't study the patients is inappropriate, because we have studied them in a number of different ways, and we have presented data on all of the groups. It's not that we have just presented data on the 128 cases. We didn't throw the others away.

We presented the data to characterize those patients in many different ways. So the sample size that we used the NPV on, you are absolutely right.

CHAIRMAN WILSON: Dr. Reller.

DR. JANOSKY: I have not finished my statement.

CHAIRMAN WILSON: Well, go ahead.

DR. JANOSKY: My statement was saying perhaps less than .4 should be used as an inclusionary criteria. So in other words that was actually the group of patients that you were looking at to answer your question of NPV, but you needed to screen quite a lot more than that.

DR. WALKER: Absolutely.

CHAIRMAN WILSON: Dr. Reller, did you want to make a comment?

DR. RELLER: Some of my earlier comments were succinct and some would even say blunt maybe, and maybe overly so. But I would like to put a different light, in terms of how I look at the decision making process that we have heard today.

I recognize how terribly difficult these patients are to take care of, and another way of looking at which standard is used against which to compare results of the EAA, CDC versus CEC, the CEC group I actually like.

If you look at it in one way, it is an evidence-based standard. You have got people taking the best available evidence they have, flawed as it may be, and coming up with a decision, and those people are very experienced.

The sort of people that you would like taking care of you if you were in Slide 2 in that unit, and they assessed 33 patients. CDC criteria put 54 patients, and 11 of them in the CDC categorization were missed, and eight in the -- if you want to look at it as an evidence-based group designation as having GRAM-negative infection.

And that is where I have my reservations of 8 out of 33, with conscientious, experienced people assessing. No one is under an illusion that they were the only ones infected, but as best as we can tell, they had an infection, and 8 out of the 33 were missed.

So that gives us -- and then coupling that with Mr. John Dawson's comments that we have a test that leaves me with facing a decision is not appreciably different from where I was as an expert evaluating these patients in the first place, the 95 percent confidence interval, with the numbers of patients involved overlapping.

So what I would do if I were to do this test is what -- and I have to paraphrase this because I don't remember the exact words. But Eric Castle in his book that was reviewed, Annals of Internal Medicine, in talking about the seduction of technology.

And that is that making clinical decisions is intrinsically making very tough ones without having all the data necessary to make them. And sometimes we order things and do tests that simply shift the ambiguity to the test from where it resided with the clinician in the first place.

And when I see something that leaves me with a probability, a likelihood, a post-test probability that is not appreciably different from where I was before, I wonder if I am not just adding something else, but I am still in the same dilemma that I was before I started.

DR. WALKER: The challenge is not in the statistics, but the challenge in the patient in front of you, and at the moment, there is nothing to change that ambiguity or that challenge in the diagnostics.

And while we now that you are absolutely right, that 8 percent of them are going to have GRAM-negative infection, and 92 percent are not, we simply don't know which of those 92 percent are not going to.

And I think that the issues of false negatives are an issue that are included clearly in the information that the clinicians would utilize. And false negatives are not uncommon in most tests in the intensive care unit.

Cultures have them, and chest x-rays have them, and therefore the utilization of this has to clearly be part of a whole armamentarium of tests, and it is new information. It is novel information. We have linked it to this particular issue with respect to a reduced likelihood of having an infection.

And it is clear that in 120 of those 128 patients that it is the absolute truth. Now, I don't disagree with any of the other statements. It's just that the challenge that we always have as clinicians is the application of statistics to the patient, to the one patient in front of you.

And while statistics deal with a hundred patients, the clinician has to deal with the patient in front of him, and so information at that point early on that might shift a -- and shift, not change, but shift a focus of particularly diagnostics, may result in better patient management, and that you may twig to something earlier on with that extra piece of information.

And we are not suggesting that it be used in isolation of other equipment. It is very important that that is not in any way being put forward.

CHAIRMAN WILSON: Dr. Charache.

DR. CHARACHE: I am going to make three comments. First, I think Dr. Janosky expressed very clearly what I was trying to drive at when we talked about the use of the word agreement.

I think you really have to be very clear of what you are agreeing to, and it has to be so specific that I think in this case we would get down to numbers that were to small to be helpful.

Secondly, I do agree -- I would like to suggest -- and Dr. Marshall indicated that perhaps the numbers that I was concerned about were not applicable. I think you are going to want to check them.

I was working from this table that you gave us, which is the CEC numbers of the positives, and the other table which we had in fact were all 10 patients, and 11 positive events in 10 patients. So they don't mesh all of the Serratias that didn't agree were X'd from this table.

DR. WALKER: We would be really happy to go over those with you. The error does not exist on that. There actually is an error in the other document.

DR. CHARACHE: But even so, there were 10 E. colis here, and there were none missed, and there are missing in other events. So I think you will just want to check on that.

DR. WALKER: Yes.

DR. CHARACHE: And then finally I think I would like to express appreciation for the fact that you, Dr. Walker, and your group have tackled an area which is as complex as this.

Ad I certainly respect the format in which you presented your data, which made it very easy to see exactly what had been done from my perspective, and as amplified here, and I do hope the panel discussion will be helpful to your group as you go forward.

DR. WALKER: Thank you very much.

CHAIRMAN WILSON: AT this point I would like to ask the FDA if they would to make any further comments, and if they have a response?

DR. GUTMAN: No. We have no further comments.

CHAIRMAN WILSON: Okay. Then let's stick to the original schedule, and let's take a break now and let's reconvene at 3:20 for the vote and recommendations.

(Whereupon, at 3:04 p.m., the meeting was recessed and resumed at 3:22 p.m.)

CHAIRMAN WILSON: At this point, it is time for the panel members to make their recommendations and final vote. And Ms. Poole will go through the voting procedures for us.

MS. POOLE: Good afternoon. The Medical Device Amendments to the Federal Food, Drug, and Cosmetic Acts, "The Act", as amended by the Safe Medical Device Act of 1990, allows the Food and Drug Administration to obtain a recommendation from an expert advisory panel on designated medical device pre-market approval applications that are filed with the agency.

The PMA must stand on its own merits, and your recommendations must be supported by safety and effectiveness data in the application, or by applicable publicly available information.

Safety is defined in the Act as a reasonable assurance based on valid scientific evidence that the probable benefits to health under conditions of the intended use outweigh any probable risk.

Effectiveness is defined as a reasonable assurance that in a significant portion of the population the use of the device for its intended uses and conditions of use when labeled will provide clinically significant results.

Your recommendation options for the vote are as follows. There are approval if there are no attached conditions. Approvable with condition. The panel may recommend that the PMA may be found approvable subject to specified conditions, such as a physician or patient education, labeling changes, or further analysis of existing data.

Prior to voting all of the conditions should be discussed by the panel. And not approvable, the panel may recommend that the PMA is not approvable if the data do not provide a reasonable assurance that the device is safe or if a reasonable assurance has not been given that the device is effective under the conditions of use prescribed, recommended or suggested in the proposed labeling.

Following the vote the chair will ask each panel member to present a brief statement outlining the reasons for their vote. Present today as voting members are Kathleen Beavis, Valerie Ng, Natalie Sanders, and only in the case of a tie, our Panel Chair, Mike Wilson.

To reach a quorum, appointed to temporary voting status pursuant to the authority granted under the Medical Device through the Advisory Committee Charter, dated October 27th, 1990, and as amended August 18th, 1999, I appoint the following persons as voting members of the Subcommittee of the Microbiology Devices Panel for the duration of this panel meeting on October 11th and 12th, 2001.

And they are Ellen J. Baron, Robert L. Danner, Frederick F. Nolte, and L. Barth Reller. For the record, these people are special government employees, and are either a consultant to this panel, or a consultant and voting members of another panel under the Medical Devices Advisory Committee.

They have undergone the customary conflict of interest review. They have reviewed the material to be considered at this meeting, and it is signed David W. Feigal, Junior, M.D., Director, Center for Devices and Radiological Help, October 10th, 2001.

CHAIRMAN WILSON: Thank you. At this point, I would entertain motions. Dr. Charache.

DR. CHARACHE: I don't think I am a voting member. Can a non-voting member make a motion or should they not?

MS. POOLE: They may not.

DR. CHARACHE: Thank you.

CHAIRMAN WILSON: Okay. So for voting we need a motion from one of the voting members of the panel. Dr. Reller.

DR. RELLER: I move that we consider this PMA non-approvable.

DR. BARON: I second.

CHAIRMAN WILSON: We have a motion and a second. Is there discussion? If not, all the voting members who are in favor voting aye?

(Ayes.)

CHAIRMAN WILSON: Those opposed?

(Ayes.)

(Vote Taken.)

CHAIRMAN WILSON: I would like each of the voting members to give the reasons for their votes, starting again with Dr. Nolte. I will start at your end.

DR. NOLTE: Basically, it boils down to the confidence that you have in the negative results in ruling out a GRAM-negative infection, and from the sample size from which we are asked to draw conclusions about that is too small.

And basically without that confidence there is very little -- it is very difficult for me to understand how this information is going to be used to change the management of patients in the ICU.

CHAIRMAN WILSON: Okay. Dr. Reller.

DR. RELLER: The request was for using this test as a rule out and I do not believe the sensitivity assessed by the various approaches taken enables one to use the test in that way.

So that it does not give added -- I don't have the confidence that it adds to the pre-test probability, and it being ruled out.

CHAIRMAN WILSON: Dr. Danner.

DR. DANNER: I don't believe that the data presented to the committee adequately gives you information that allows you to interpret this test appropriately, and to change any kind of clinical decision or management of patients.

CHAIRMAN WILSON: Thank you. Dr. Beavis.

DR. BEAVIS: I do not believe that the data that we received showed clinical effectiveness. That is, that the results would provide clinically significant results that would make a change in the patient care rendered.

CHAIRMAN WILSON: Okay. Dr. Ng.

DR. NG: I believe that the data as presented in fact showed that the strength of the negative predictive value was in fact directly related to the low prevalence of GRAM-negative infections. I see no clinical role of this test in clinical management.

I also feel that the neglect of the importance placed on the sensitivity was a failing in that there is great importance attached to missing one out of five GRAM-negative infections with this test.

CHAIRMAN WILSON: Dr. Sanders.

DR. SANDERS: I had concerns about the safety of the test and that clinicians may rely upon a negative result as an indication to alter therapy and may not take into consideration other pieces of information that might be of benefit to the patient.

CHAIRMAN WILSON: And Dr. Baron.

DR. BARON: I would like to say that I think that this test could be a very useful test in a research setting, which would not necessarily require FDA approval. But that for a clinical laboratory that the test would not significantly add diagnostic failure to clinicians.

CHAIRMAN WILSON: Are there any comments that any of the other members of the panel would like to make at this time? If not, Dr. Gutman, any comments from the FDA?

DR. GUTMAN: No.

CHAIRMAN WILSON: Okay. That will conclude this part of the meeting. I would like to thank all of the members of the panel for their time and effort today, and I would also like to particularly thank the sponsor for all the work that they had done in the presentation today.

We do have to break now. We have go give the next sponsor time to get set up. We are going to try to reconvene if at all possible at four o'clock. Thank you.

DR. GUTMAN: Can I ask before we recess if we could go around and ask the panel members for their advice on what might be done to make it approvable?

CHAIRMAN WILSON: Sure. That would be fine. Let's start with -- Dr. Janosky, do you want to start?

DR. JANOSKY: Most of the issues that were brought up today I think could be addressed, and they could be addressed using them in the design of the study. In particular, some of the issues that should be paid attention to would be the differences among patients, and getting a fair enough sample.

I understand how difficult that can be, to look at differences either across organisms or across sites, or by personal characteristics, or by prevalence at different sites, just to show that there is something, irrespective of what is going on in some of the other issues. But that would be the one that I would concentrate on.

CHAIRMAN WILSON: Dr. Nolte.

DR. NOLTE: Basically, it is a tough issue for all the reasons that have been talked about here. I mean, really it boils down to whether we are talking about building a better test for endotoxin, and I think the sponsors have done that.

It really boils down to what that test means in an ICU patient population, and equating the presence of endotoxemia with infection. And I have heard a number of experts, and I by no means am an expert on taking care of ICU patients.

But I have heard a number of you talk about that today, and that is not a direct equation. That is not -- you know, X doesn't equal Y. So you really have to reexamine the whole paradigm in terms of how you put together a clinical trial in order to convince a diverse panel like this of the value of an endotoxin test in this setting.

CHAIRMAN WILSON: Dr. Reller.

DR. RELLER: There have been many things mentioned earlier and I don't have any further suggestions.

CHAIRMAN WILSON: Dr. Danner.

DR. DANNER: Well, although I applaud the effort of the company, and I would agree with Phil Dellinger, who now has had to leave, that this is an unmet need and something that would be useful if there were such a test.

I am concerned that this test is not that test, and that no matter how you test this technology that you are going to keep hitting up against the same limits of it.

So I guess if you -- I would advise you not to pursue making or trying to make this approvable because I am not sure that it can be, or I am actually reasonably sure that it can't be.

But if you were, you would need to show that it was clinically relevant to having this data impacted positively on patient care and improve patient care in the ICU.

I think that is a very tall order. It would require a very large study and I think even with the correct numbers I would be very concerned that it just wouldn't pan out.

I also would add that one of your comments earlier about people having converted their tests to positive when they smoke makes me concerned that perhaps you are not always measuring endotoxin, because I don't smoke, but I know a lot of people who do, and they don't get fever when they smoke.

And your test is sensitive at the picogram level and people are like rabbits, and tiny, tiny doses of endotoxin give them fever. So if your test is detecting endotoxemia during smoking, I am concerned that it is detecting something else other than that.

CHAIRMAN WILSON: Dr. Beavis.

DR. BEAVIS: I don't have anything else to add. Thank you.

CHAIRMAN WILSON: Dr. Ng.

DR. NG: I would like to agree with Dr. Danner. I think you have an excellent assay. I think the problem is that the physiologic variables are going to handicap it, and I don't think you can ever overcome those with however you design a future study.

CHAIRMAN WILSON: Dr. Carroll.

DR. CARROLL: I agree with other panelists' comments, but in particular I think the nature of testing for endotoxemia is just very difficult, and I just want to reiterate what has already been said about that.

CHAIRMAN WILSON: Dr. Sanders.

DR. SANDERS: Well, I thought that this was very ambitious and was actually looking very forward to this discussion, because if we could have a test that would allow us to reduce our use of very ototoxic, nephrotoxic, and hepatotoxic drugs on very sick people, and reduce the cost of their care, and shorten their ICU stays, that would be very wonderful.

However, I wasn't convinced that this particular product at this time, given the low prevalence, and even the changing nature of toxemia in the ICU, was the product that would allow us to do that.

CHAIRMAN WILSON: Dr. Baron.

DR. BARON: Well, I don't know if this would work, but maybe if you limited the scope to a certain kind of infection, sepsis, or something where you could fine-tune the test a little bit better than just taking all-comers into the ICU, the data might end up to prove more correlative.

CHAIRMAN WILSON: Dr. Nachamkin.

DR. NACHAMKIN: I agree with Ellen Jo that if you perform a larger study and increase those numbers of specific infections -- pneumonia, bacteremia, et cetera -- that you might be able -- and again you would have to wait for the data, but you might find some better correlation of your test, and the ability to rule out a certain type of infection.

So that is the only situation that I see where further development might be warranted. But if it is just going to be applied to just the general population, I agree with the rest of the panel, and that I am not confident that they are going to go very far with that.

CHAIRMAN WILSON: Dr. Charache.

DR. CHARACHE: I also feel that you have taken an extraordinarily difficult group of patients to try to sort out with an extremely sensitive assay, and I am not certain that that is a population that is going to prove rewarding.

At the same time I am intrigued with the chemistry that you are employing, and I am wondering if it might not be helpful to look at some of your false positives, and see where or what the cause of them might be.

And whether the technology might not be extremely valuable if applied in a slightly different manner. I am wondering about the excitation of the complement pathway that you may be looking at, or whatever else it is that is giving you the signal that you are receiving.

And I might look at some patients who have that type of activity going on, like a lupus patient, or whatever, and look for your false positives, where you can't say, well, maybe this patient has endotoxin from the GI tract, and maybe I am measuring something that is not endotoxin.

But perhaps working it through some of the discrepant results might be a clue on how to solve and clean up the assay.

CHAIRMAN WILSON: Mr. Reynolds.

MR. REYNOLDS: When I looked at your initial package, one of the things that intrigued me was in your cross-reactivity study, the negative that you got with vibrio cholerae.

In looking at your data, it seemed that there are certain groups of organisms that tended to give you negatives. And I am just wondering if you have really looked at those false negatives or done more work with vibrio cholerae to pinpoint what causes a negative test.

Because if you clean that up, and eliminate those false negatives, I think you have might a useable test.

CHAIRMAN WILSON: And Dr. Durack.

DR. DURACK: Well, certainly from the point of view of an infectious diseases clinician, I would be very happy if you succeeded in the future. Just four points to what you heard. I certainly would advise resolving the negative predictive value gold standard issue before going forward and to find an acceptable way of handling the gold standard issue.

I think you could relook at the cutoff and make sure that you do have the best cutoff, and whether the .3 would be a better cutoff. And increase the numbers and look at the subgroups, and perhaps define value in one important, or more than one, important subgroups.

And finally define a way to demonstrate how a clinician in practice would use the result in a way that would add value to the clinical decision making.

CHAIRMAN WILSON: Thank you.

DR. GUTMAN: Thank you very much.

CHAIRMAN WILSON: Thank you. Again, we will try to reconvene as close to four o'clock as we can.

(Whereupon, at 3:42 p.m., the meeting was recessed, and resumed at 4:07 p.m.)

CHAIRMAN WILSON: Okay. At this point, I would like to reconvene the meeting. The next item on the agenda is new business, and I would like to remind everyone that this is a pre-market notification, also known as a 510(k) submission, that is being brought before the Panel today.

The FDA is going to ask for recommendations and advice, and there will be no final vote on a 510(k) submission. This pre-market notification submission is for a in vitro diagnostic device for detective and measuring urinary tract infection by semi-quantitative analysis of volatile compounds released from urine samples.

I would like to ask the panel to hold their questions until after the initial three presentations from the sponsor, and I would also like to remind the audience that only panel members can ask questions of the speakers.

If the sponsor is ready, I would like Mr. James White to give the initial introduction.

MR. WHITE: Thank you. I would like to thank the FDA and this gentleman here, and Members of the Panel, for inviting us here today. What I would like to do is go through the Osmetech team here, and then talk a little bit more about the clinical investigation that we have here, and then go through the agenda.

My names is James White, and I am the CO of Osmetech, accompanied by David Grindrod, who is our chief operating officer; and John Plant, who is the project leader of the urinary tract infections work that we have been doing.

And he has been working on this for the last three years, and has done the day to day work with both the FDA and also some of the clinicians that we have been working with.

We also have Paul Travers, and he has had around 12 years experience with conducting polymer technology that we used, and has been instrumental in taking it from its initial university background and beginnings really to the commercial product that we are about to discuss today.

The clinical investigators that we have used on the vapor performance and reproduced work, in terms of performance studies, we have been working with Gary French, who is the head of clinical microbiology at St. Thomas' Hospital in London.

And Patrick Murray, from Baltimore and the University of Maryland, who will present to the panel today the clinical studies and also the conclusions.

Andrew Onderdonk, who has been working with Brigham and Women's Hospital, and he has worked with us on the performance and reproducibility studies.

Andy has been working with the company for the last five years, and has been instrumental in taking us from the industrial company that we started as, and through to the medical diagnostic that we are focused on today.

In terms of the agenda, I will give a quick overview of the company, and also the regulated history. John Plant will talk about the device description, and within that a little bit more about the technology. And also some of the studies that we have done, prior pivotal studies.

And then Patrick Murray will go on to talk about the conclusions, and then I will field questions after this as well.

The company was set up as AromaScan back in 1993 from some technology from the University of Manchester Institute of Science and Technology, in England, and the founder of the technology is a gentleman that we still work with very closely today.

Back in those days the company was very much focused around industrial applications, but back in 1998, we really changed to reflect a move away from being an analytical instrument company to a health care diagnostics organization.

In terms of the regulatory history, we started talking with the FDA back in January of 2000. I had a number of very helpful meetings and talking about clinical protocols, and certainly some of the intended uses that we would like to think that the technology would be used for in the health care area.

The main conclusions really from the conversations that we had were that it firmed up the regulated pathway, in terms of 510(k) for the clinical pivotal files, and it also confirmed the number of study sites we would be using for the performance trial, which was three sites; and the reproducibility study, we would be using two sites.

And also there was a confirmation that the UriscreenTM would be our predicate product, which is similar in terms that it is an indirect test. However, there are a couple of differences beyond that, in terms of we are an automated device for clinical laboratories; whereas, there is a home test, which is a manual test.

We finished our performance and reproducibility studies towards the beginning of this year, 2001, and then submitted the 510(k) in April. And really between April and August of this year, we have been fielding a number of questions, and have got all the answers back to that.

And really what we would like to do today is set out through the presentation that both John Plant and Pat Murray will give, is really some of the responses to those questions; and also the other four questions that the FDA have posed to us.

And really the presentation, plus the appendix that we have attached to that, hopefully should go through some of the answers of that for you. So at this stage, I would like to pass on to John Plant.

MR. PLANT: Thank you, James. My name is John Plant, and I am the health care UTI project team leader employed by Osmetech. I would like to start the presentation of the OMA-UTI device description by looking at the intended use statements.

The Osmetech OMA-UTI instrument is an automated in vitro diagnostic device intended for use by clinical laboratory health care professionals as an aid to the detection of bacteria associated with urinary tract infections.

The OMA-UTI indirectly measures bacterial infection by semi-quantitative analysis of volatile compounds into the headspace above a urine sample.

The OMA-UTI is a screening device intended to reduce the need for unnecessary culture.

The OMA-UTI device is not a substitute for culture since it does not identify the organisms present. The next slide, please. The OMA-UTI device measures the presence of bacteria indirectly by detecting volatile bacterial metabolites from the headspace above the urine samples.

The technique is semi-quantitative, giving a positive or negative results at the threshold of 1 times 10 to the 5 colony forming units per Ml as determined by -- and to the left is a photograph of the OMA-UTI device.

I will walk or talk through a typical analysis sequence in a few moments. However, briefly, the operator's interaction with the device is to load the samples, the critical samples into the carousel, and then load the sample codes by the keyboard and start the system.

After that the rest of the system is fully automated. The diagram on the right-hand side is a line drawing of the OMA-UTI instrument, with the covers removed and it just shows a bit more detail of the specific parts of the instrument.

Essentially because we are delivering a heads space from the sample, the whole of the unit as you can see there can be reduced to the sample vial containing the urine.

A needle, which delivers humidified gas into the sample and displaces the head-space through a transfer line, and then to our sensor technology. The rest of it as you can see is to automate that process and to control it.

Two other points to make is that the sensor is housed in a temperature controlled environment, and which prevents environmental changes in temperature affecting the sensor response.

Secondly, the gas that is delivered through the sample is humidified, again to eliminate the environmental effects on the sensor. The software controls the correct operation of the device.

And it checks the temperatures, and the flow rates, and the humidities which are all monitored, and should react as to the specifications. If not, the system shuts down. The system is designed to fail-safe in the event of a failure.

If you look at the Osmetech technology, this is an example of the sensor, which is the heart of the system. The diagram or the picture on the left-hand side shows just a small segment of this array.

The sensor array is an array of four different polymer types, which then repeats across the whole array. The three black squares that you can see on the photograph are the sensors themselves.

The management process is made by applying the voltage to the sensor by the gold electrodes. And as you can see the gold electrodes is at the top of the sensor, and then also the wire bottoms, which take it on to the ceramic substrate that we use.

The voltage is applied and the change in resistance is measured as the sensors are exposed to the urine head-space. The resistance of the sensors change depending on what is absorbed on to the surface of the sensor.

The chart on the right shows the four polymers responding when exposed to the culture sample. The sensor is exposed for three minutes, and so the section that you can see here with the two sensors are strongly responding is where it is exposed to the sample.

The output from the sensors is processed using principal components analysis to give either positive results or negative results. The next slide, please.

During the UTI it uses controlled chemicals, and these are the same chemicals that we have identified as the volatile metabolites and the bacteria that is associated with the UTI. We use both negative and positive controls.

Once the new sensor is put into the device, a reference run is performed using in the factory, or if on site, by a Osmetech service engineer.

This sets up the principal component reference map for that particular sensor, and also it checks the algorithm with using an algorithm that the sensor has sufficient sensitivity and sufficient performance to do the job.

After the reference map has been made, the calibration is then run by either a Osmetech service engineer or it can be run by the clinical lab supervisor.

The calibration procedure sets the classification thresholds, and then a sample giving a responsibility classification threshold, which above is reported as positive and below it is reported as negative.

And that sets up the configuration for the system to be used by the operator. The operator must perform a system check using the same control chemicals prior to every sample batch to ensure suitable performance of the system.

And once again, when the sample batch is finished, then the operator must then return to a further system check before running any future sample batches.

This slide shows a typical sampling sequence. The samples arrive in the clinical laboratory and are stored at 2 to 8 Celsius. We have conducted studies on the untreated samples to show that they have 24 hour stability at 2 to 8 Celsius. There is actually stability of the metabolites in the sample as has been demonstrated.

Once there is a sufficient batch of samples to run the operator prepares one mil into the Osmetech vial containing additives. These additives, the acid and salt, promote the release of the metabolites into the head-space.

The operator then loads the carousel, and inputs the sample codes, and from then on the sampling is automated. Currently the first results are available within 6 hours.

Again, we have conducted studies on the treated samples to ensure that stability is sufficient for a full carousel run. At the end of the batch a report of the results is printed out. Next slide, please, David.

Summarizing the studies that have been conducted in support of the 510(k) submission, there has been a proof of principle study conducted at St. Thomas' Hospital in London, the U.K.

And during the principal study the presence and absence of blood in the urine, and specific gravity of the urine, were both measured and shown not to effect the Osmetech results.

Further, in-house branch testing using a water matrix looked at nine compounds covering urine and their effects on the OMA-UTI results, non-inter-fed with the OMA-UTI's ability to detect positive samples.

However, there was a suggestion from the dates that sodium nitrate enhanced the sensory response. There have been two clinical studies of the OMA-UTI to look at device performance and reproducibility, and Dr. Murray will take you through those now.

DR. MURRAY: He never lets me keep the toys that he has. I would like to thank the panel, the FDA panel, for the invitation to present the clinical studies that I was able to participate in. If we could have the next slide, please, David.

There are two objectives of the clinical studies that we performed. The first one for the first study was to evaluate the performance characteristics of the OMA-UTI system, and to compare that with standard microbiologic culture, and I will define that in a second.

That was considered our gold standard, and then we also compared the performance of the OMA-UTI system with the predicate device that the FDA selected, and I will present some of that data in a second.

The second objective was as part of the reproducibility studies to look at inter-site reproducibility of the system. The next slide, please.

The design of the study was developed in collaboration between the FDA and Osmetech, and what I have done here is summarize some of the important points of the study design.

Informed consent was not sought for any of the urine samples that were processed in this study, and the reason for that was that we wanted to collect consecutive urine samples and not introduce a bias in the types of samples that were being analyzed.

Samples were not screened for any medication, including antibiotics, and the reason for that is that we recognized that the reports of the presence of antibiotics on requisitions that were submitted with the sample would be unreliable and so that we would have had to review the medical charts.

And again since we did not have informed consent, we couldn't do that. So we recognized the fact that if antibiotics were present, and since we are measuring a metabolic byproduct of an organism, the presence of antibiotics could bias against the performance of this system, and that was accepted as part of the study design.

Samples containing preservatives, such as boric acid, were excluded from the study. They are obviously easy to identify, and it was recognized that if you had an inhibitor present that we would anticipate that the samples would be negative.

And there is no claim that the system could work with samples in the presence of boric acid. The demographics of the population that was studies was comprehensive. As I said, we did not exclude any patient population.

And so samples were collected from the emergency department, and from the various clinics in the medical centers, from the intensive care units, and from general surgery and medicine floors.

The confirmatory test was the standard urine culture, and the definition for a positive specimen was the presence of at least one organism and concentrations of 10 to the 5 organisms per ml or greater.

We recognized again that if you had a mixture of organisms and if the composite was greater than 10 to the 5, we could anticipate that we would have some positives with this assay.

Again, by definition, those specimens, since no one organism was greater than 10 to the 5 organisms, they were defined as being negative. Finally, the patient treatment, or any management of the patient, was not influenced by the results of the OMA-UTI test.

Again, these results are not reported to the physicians. We were processing excess urine that was submitted with a routine urine culture, and so again patient management was not influenced. Next slide, please.

As has already been indicated, there are three centers that participated in the perform study, the first study that was performed. Dr. Gary French, at St. Thomas' Hospital in London; Dr. Andrew Onderdonk at Brigham and Women's Hospital in Boston; and myself at the University of Maryland Medical System, in Baltimore.

A total of 1,038 samples were evaluated, and let me present the data for those samples in this slide here. Of the 1,038 samples that were submitted, there is a total of 147 samples that were culture positive, and that is roughly 14 percent of the samples that were submitted were culture positive.

And we feel that is representative of most studies that have looked at a general population of patients. We had 891 samples, or 86 percent, that were culture negative by the definition that I gave.

If we look at the sensitivity of this test, 119 samples were OMA-UTI positive of 270 samples that were -- I'm sorry, but 119 samples were positive of the 147 samples that were culture positive, or the sensitivity was 81 percent.

The specificity was 83 percent. That is, 740 samples were OMA-UTI negative of the 891 that were culture negative. The overall accuracy of the test, that is, where we correctly identified both the culture positive and the culture negative samples, the overall accuracy was 83 percent in this study.

The negative predictive value was 96 percent. That is, 740 of the 768 OMA-UTI negative samples were culture negative; and the positive predictive value was 44 percent, or 119 of the 270. Next slide, please.

What I would like to do is to further examine the tests where we had both false positive test results and false negative test results. There is a total of 151 false positive test results that were analyzed.

And again remember that we have defined the culture as being negative if there is no single organism that was greater than 10 to the 5 organisms per milliliter.

When we analyzed the false positive results, we found that approximately half of the results were associated with multiple organisms being present in culture.

We found that there were 11 specimens that had a single organism present and culture in that organism by definition had to be less than 10 to the 5 colony forming units per milliliter; and with 66 samples, we found no organism was present in culture.

For the 28 false negative tests, when we analyzed those results -- and it has to be again pointed out that there is no assessment of antibiotic use, which we would anticipate in the presence of antibiotics that this test would not perform as well as in the absence of antibiotics.

But also because we did not review the clinical records, there is no assessment of the clinical significance of some of the organisms that were present in concentrations greater than 10 to the 5.

And as an example, we had a number of organisms that by most definitions would most likely be clinically insignificant as the cause of urinary tract infection.

And these include -- there was one isolates of corynebacterium, and one isolate of coagulase-negative staph lococci; and two islets of lactobacillus; and four islets of enterococci. They were all present in large numbers in culture and were not detected by the OMA-UTI system. Next slide, please.

What I have done in this slide is compare the performance of the OMA-UTI with some predicate devices, and what we have listed here in the first column is the statistical data for the OMA-UTI system, and I have already reviewed that.

And the second system there is the Bactis 160 system, or the Combact System. That system detects microbial presence by labeling the organisms with a fluorescent dye.

And concentrating them on a filter, and then scanning that filter or counting the number of particles that are present on the filter, and then making an estimate of the number of organisms in the urine sample. So that is not a system dependent on growth of organisms or the metabolism of organisms.

The last three systems that were evaluated were all dependent on enzymatic activity, and they are all constitutive enzymes present either in the organisms or in the cellular material that may also be present in the urine.

The first system is the Uri-Screen, which is a system that measures catalase activity; and again that could be catalase activity present in organisms, in leukocytes, or in squamous epithelial cells that may have contained the urine sample.

The other two systems that I have listed on this slide were -- it is data that was presented to the FDA with the Uriscreen data as the predicate devices for the Uriscreen system. So that's why some of the numbers are -- that the number of samples are the same there.

The Multistix Reagent Strip measures leukocyte esterase, or the presence of leukocyte esterases, which is obviously not an enzyme in bacteria, but rather associated with the leukocytes that may be present in an infection.

And the last system is the nitrate reductase test, which again measures an enzyme produced by many common bacterias, such as urinary tract infections.

The clinical trials, I think it is important to recognize that there were some differences. As I have indicated in the study that I presented here on the OMA-UTI system, consecutive urines were selected and the same was done for the Bactis system.

In contrast, the other three asymptomatic tests selected only symptomatic patients. And so as an example, if you had a test for leukocyte esterases, and you are essentially measuring inflammation, then you would expect that an inflammation would be more common in systematically infected patients, as opposed to asymptomatically, but significantly infected, patients.

So I think there is a significant difference in study population for some of these studies that we are looking at. In each of the studies, with the exception of the Bactis system, three sites participated in the clinical evaluation.

So they are essentially the same, and in all five studies that are presented here the definition of a positive urine culture is the same here.

The number of samples are on the next row, and you can see that the asymptomatic test had relatively few samples that were evaluated. There was a large of samples evaluated with the Bactis system and a reasonably large number evaluated with the OMA-UTI system.

The sensitivity as I have already indicated was 81 percent for the OMA-UTI system; and it is slightly less than what we see for the Bacis system, and the Uriscreen system and the leukocyte esterase system, and significantly greater than what we see for the nitrate reductase test.

The specificity is 83 percent for the OMA-UTI system, which is comparable with what was seen with the Bactis system, and superior to what is seen with the catalase test and leukocyte esterase test; and much less than what is seen with the nitrate reductase test.

And I think that is sort of an interesting observation. If you think about it and use the example of the nitrate reductase test, it is a relatively insensitive test, and that is well recognized in published reports in the literature.

If you have an insensitive test, then you would expect that your sensitivity obviously is going to be low, but your specificity, that is, calling samples negative, should be high and that is exactly what you see there.

So maybe a more reasonable assay or statistic to analyze is the accuracy, and that is the bottom row, and for the OMA-UTI system the accuracy was 83 percent.

Data was not available for the Bactis system, but looking at the numbers that are presented, we would estimate that it should be comparable to the OMA-UTI system.

The problem is that we don't know what the prevalent disease is, and so we can't make those calculations. The accuracy for the catalase test and for the nitrate reductase test is essentially identical to the OMA-UTI; and the accuracy of the leukocyte esterase test is significantly lower than what was seen with the other systems. Next slide.

The second study that was performed was the reproducibility study, and again it was performed in two studies, the Boston center and the Baltimore center.

Samples in this study were pre-screened by microscopy to select for a higher proportion of positive cultures, and the reason for that was that if we were looking at reproducibility, and we analyze that 86 percent of our samples are negative, I think it is not very useful to say that we have a very reproducible test with negative samples.

We wanted to also look at how reproducible the assay was with positive samples, and this was -- this modification of the protocol was discussed with the FDA.

The samples when they were collected in the individual laboratories were divided into two aliquots. One aliquot was refrigerated, and the second aliquot was sent to the companion laboratory, and obviously since that is an overnight shipment, and so each site tested all samples 24 hours after collection and after they had been refrigerated for 24 hours.

So the testing that was done in the Baltimore laboratory was being done at the same time as the testing that was being done in the Boston laboratory.

A total of 249 samples were run, and 85 were positive or roughly 35 percent of the samples were positive, and 164 were negative. There was 93 percent agreement between the two sides for the study results, and the Kappa statistic assessing the strength of that agreement was .86 percent.

And as indicated on this slide, based on the analysis and Kappa statistics, that would be considered a very good strength of agreement. Next slide, please.

So, in conclusion, we felt that we met the objectives of the study. The assay was accurate and had an accuracy of 83 percent. It was substantially equivalent to the predicate device and three other devices that were analyzed that are in common use.

And we found that the testing was reproducible; and that 93 percent of the assay results yielded the same results in two laboratories. Next slide.

One possible clinical paradigm on how this system could be used is that if there is a high index of suspicion that the patient had a urinary tract infection, that is, if the patient was symptomatic, we feel that in that situation it would be appropriate to culture the patient and not delay processing by doing some sort of a screening or accessory test.

If there is a low index of suspicion, and let's say you are screening the population of diabetic patients, or asymptomatic patients, then it could be appropriate to use this test.

And if the test results were negative with a high negative predictive value, the testing could stop at that point. And if the testing was positive, it would be appropriate to culture the sample.

With that, let me stop, and I will turn it back to James White.

MR. WHITE: Thanks, Pat. At this stage would you like to see questions? If you would direct them to me, and then I will pass them to the maybe more appropriate people that we have with us here today.

CHAIRMAN WILSON: Okay. Dr. Nachamkin.

DR. NACHAMKIN: Am I correct in that to do this test the samples would have to be refrigerated during the transport to the laboratory?

MR. WHITE: I will pass that over to John Plant.

DR. NACHAMKIN: And then along with that, many laboratories do a lot of their urine cultures from samples coming from off-site from outpatient clinics or whatever, frequently in preservative.

So boric acids are a very commonly used method of transport, particularly when you are going to be doing cultures. So those are two questions.

MR. WHITE: John, the questions were -- let me kind of play them back -- to the samples need to be kind of transported in refrigeration; and also given that there are a number of samples that are transported in preservatives like boric acid, what is the impact on that?

MR. PLANT: Well, firstly, there is no requirements for storing the samples refrigerated to the clinical lab. And secondly we have labeled the device that samples in boric acids are not to be used.

DR. NACHAMKIN: So with regard to refrigeration, you said that the urine is stable under refrigeration for up to 24 hours. What is the time interval from the time that it is collected to when you advise that it be tested without refrigeration?

MR. WHITE: Let me summarize that. Well, Pat.

DR. MURRAY: I am not really sure that we really can completely answer your question for logical reasons, and that is that as you know we would not leave a urine that is going to be cultured at room temperature for a significant period of time.

We do know that in specimens that were sent to the laboratory, where there can be a delay of two hours or even more than that, that the assay performed well.

And there is data that I guess John could share on stability beyond that when it is refrigerated, but none of us would recommend holding urines for long periods of time before it is processed.

The way the study was designed was that urines would be submitted to the laboratory, and we would go ahead and do our routine cultures, and we would set those aside then in the refrigerator and batch them and do the testing with the sample.

CHAIRMAN WILSON: Dr. Durack.

DR. DURACK: A question I think for Dr. Murray. I may have missed it, but did you include yeasts in the positive, or were yeasts excluded? Did we learn anything if they were included?

DR. MURRAY: We included all organisms that were greater than 10 to the 5, and we have a slide -- David, can we show that slide? I can answer that maybe when we see numbers a little bit better.

This slide here is a listing of all of the organisms that were greater than 10 to the 5 in the performance study. And as you can see at the bottom of the slide, there were 10 yeasts that were detected there.

Of those 10 yeast, 5 were detected with the system. So they would recognize that the system is not as sensitive for yeast. Now, because of the small numbers, statistical analysis didn't demonstrate that there was -- that this difference was statistically significant.

But I think inherently that it didn't perform as well with yeast samples, and that could be just the selection of sensors that were used. I should also point out that there is no claim for yeast. The claim, if I am not mistaken, is for the detection of bacteria.

DR. DURACK: Right. And were there any of the bacterial subgroups that showed any unusual difference from the standard sensitivity?

DR. MURRAY: The next slide I think would probably address that. You can sort of scan down the list and see that there is really a scattering. The largest number that were not detected by the OMA-UTI was in Escherichia, 10 of the 71 strains were not detected.

If you do the statistics that is a sensitivity of about 85 percent, or slightly higher than the overall sensitivity of the system. But in the statistical analysis there really wasn't any difference. There wasn't any one organism that clearly failed to be detected.

DR. DURACK: Thank you.

DR. MURRAY: Irv, you had a second question and I can't remember what it was, but I was going to address that.

DR. NACHAMKIN: Well, I was just a little concerned about the boric acid issue, only because -- and thinking now in my own laboratory, we get all of our outpatient urines in boric acid. So in order to use a test like this, I would have to now switch entirely to non-boric acid.

DR. MURRAY: Well, the bottom line is either you switch and eliminate boric acid and use the test; or you don't switch and use boric acid, and you don't use the test. They are not claiming that this system will work with boric acid, and you wouldn't expect them to.

DR. NACHAMKIN: Right. Are there any other preservatives that will preserve the culture integrity of the urine that might work in this?

MR. WHITE: Could I get Paul Travers to talk about the boric acid.

CHAIRMAN WILSON: Could you introduce yourself, please.

MR. TRAVERS: I am Paul Travers, and I am the sensor development team leader for Osmetech. When we make the decision to exclude samples that have been stored in boric acid for the trial, we have some preliminary information which suggests that the boric acid might interfere with the assay.

To test that, we actually include the boric acid as one of the interfering substances in our bench testing of interfering substances trial. And in that particular study the boric acid did actually interfere with the assay.

DR. NACHAMKIN: It did not?

MR. TRAVERS: It did not. It did not interfere with the assay of pseudo-samples, which is what we could prepare on the bench. So we were cautious because of this preliminary evidence that it looked like it might be a problem. But subsequently it didn't appear to be a problem.

DR. NACHAMKIN: So it is possible that if you were doing another trial using boric acid in transporting the urine that it might work?

MR. TRAVERS: Yes, I believe so.

CHAIRMAN WILSON: Thanks, Paul. Dr. Carroll.

DR. CARROLL: Yes. I am a little confused by the Group B Strep issue. I think in the information that was provided to us that it says that you had not really studied volatile gases emitted from Group B Strep. Yet, you have some clinical data on those.

And this relates back to your clinical paradigm. One of the groups or patient populations that may be screened for asymptomatic bacteria is the pregnant female.

Often times pregnant women with Group B Strep urinary tract infections are not symptomatic. So could you just clarify whether this will reliably detect Group B Strep or not.

We only have three isolates up there and so I think it is difficult to make that determination from the clinical data.

MR. WHITE: Can I get Andy Onderdonk to come and talk to that one for you, in terms of the Group B Strep.

CHAIRMAN WILSON: Could you introduce yourself, please.

DR. ONDERDONK: Yes. My name is Andy Onderdonk, and I am the Director of Microbiology at the Brigham and Women's Hospital. Although we did not study that group specifically, you know, because there was not consent, obviously we received samples, at least at Brigham, and I am sure that some of those were women being screened.

The volatile compounds that this system detects are produced by Group B Step. So one would anticipate that that organism should be detectable with this system, and I think the minimal data that you have here, where you have 3 and 2 were detected, speaks to that point.

CHAIRMAN WILSON: Dr. Nolte.

DR. NOLTE: What are the volatile compounds that are being detected?

DR. ONDERDONK: I will just let the CEO answer that one. That is proprietary information, I think.

MR. WHITE: Andy is correct, and that the volatile metabolites that we are detecting are proprietary, in terms that there are a number of key ones which are given off, and that surely is the basis of the test.

DR. NOLTE: The other part of that question is that one of the other speakers alluded to the fact that patients -- there was some concern about antibiotic therapy influencing the outcome of the test results.

Is there any reason to think that issues are any different for culture as they are for this system? I mean, are you detecting volatile metabolites as a result of the organism growing in the urine? Help me to understand what we are detecting.

DR. MURRAY: In this system, you are detecting organisms that are being produced by the metabolic activity of the organism.

DR. NOLTE: So if organisms are growing, then they are producing metabolites?

DR. MURRAY: I think we all have had the experience when we look at a urine culture that the area where the urine was initially inoculated there is no growth, and it is when you streak away from that area that you do get growth. And in those types of urine specimens we make an estimate, and not based on the total number of organisms, but an estimate of what the total number would be from that plate.

And you can have the center of the plate has no growth because there is still antibiotics there, and you have growth and is quite heavy. Well, you know that is greater than 10 to the 5.

In this system, because the antibiotics remain in contact with the organisms, the organisms will stop metabolizing and you would expect that until the antibiotics are removed it is going to affect the results of the test.

But again it is something that we can theoretically address, but until you actually look at the antibiotics the patients are receiving, and look at the performance of the test, you can't verify that the antibiotics are affecting it. But I would certainly assume that it would.

CHAIRMAN WILSON: Dr. Beavis.

DR. BEAVIS: Dr. Murray, I don't think you want to go far. I had a couple of questions for the data that you presented and one was from the chart, titled, "Performance Characteristics."

And on that chart you were reviewing the false negative tests, the 28 specimens that were false negative. And you say that there was no assessment of clinical significance, and then you list eight organisms.

And the organisms -- you listed four organisms from eight different specimens, and the four organisms that you listed were the coryne bacterium, the coagulase-negative staph, lactobacillus, and the enterococcus.

What about from the other 20 specimens? Were they also organisms that we would typically think of as skin flora?

DR. MURRAY: No. Most of those organisms would have been ones that we would consider a uropathogen. Whether they were truly significant uropathogens there, or organisms that had been present in the urethra and contaminated the specimen, and then grown during the glazing and processing we don't know.

That would have been a clinical assessment.

DR. BEAVIS: But it would have grown greater than 10 to the 5th?

DR. MURRAY: Yes, that's correct. That's why the performance of the OMA-UTI system was considered a false-negative. We defined all positives based on the culture itself.

DR. BEAVIS: So you could say then that of the 28 that you missed, eight of these weren't what we typically think of as skin flora without having to do the clinical chart review. But the other 20 were ones that we more typically think of as uropathogens?

DR. MURRAY: Right.

DR. BEAVIS: Okay. And I had another question --

DR. MURRAY: And also -- I'm sorry.

DR. BEAVIS: No, go ahead, if you wanted to clarify or --

DR. MURRAY: No.

DR. BEAVIS: Okay. And I had a question from your next chart. It was the table on that chart labeled performance characteristics and clinical comparison to culture.

And this is something that Mr. Plant said, and then I think you elaborated on it, which was with the intended use of this test is. Mr. Plant said that it is to reduce the need of unnecessary cultures, and it is not a substitute for culture.

And then I think you had mentioned that for the positives that one would want to culture to be able to identify and so forth. So if one wants to be able to detect the positive cultures by this system so that they could then be plated out, the sensitivity of this is 81 percent, but the Uriscreen is 95 percent.

And I am bringing this up because even of the accuracy of the two, and that is when you add the ones that are in agreement as to the true positives, as well as the true negatives, they are. You know, the agreement is there.

But I guess I view this device as more of a screening device, and in that situation the sensitivity seems to be of a bit more importance. I was hoping that you could clarify that for me.

DR. MURRAY: Most people would consider the negative predictive value to be the most useful, so that you could eliminate negative cultures. And the paradigm that we shared at the end of the presentation said that if there is a strong suspicion that the patient has an infection, at least I don't believe the specimen should be screened.

I think that if there is a strong suspicion that the specimen should be processed. So what we are really looking at are the large number of specimens that we all receive in our laboratory would be small, where there is a small index of suspicion that there is disease, but to still submit those specimens.

But what we would like to be able to do by any screening system, or by any aid, is to eliminate as many of the negative ones as you can, recognizing that you will be culturing -- if the test is not highly specific, you will culture excess numbers of specimens.

DR. BEAVIS: Yes, I guess my thought is that I would rather culture extra specimens that are going to be culture negative than miss some that are positive.

DR. MURRAY: Right. It really gets back to your first question, and I had started to make a comment and then decided that I would wait a second. And that was that an additional five of those specimens as we have already talked about was with the yeast that were missed.

And again Osmetech has not claimed that with the yeast with the sensors that are being evaluated here. So the overall sensitivity -- again, we showed the chart with the performance characteristics compared to the other ones, and ultimately what you are asking is the overall sensitivity of the tests.

And the overall sensitivity of the tests is not as good as some of the tests that have been approved, and you can look at the Bactis system or the Uriscreen system has you have pointed out.

On the other hand a very common test that is used s the nitrate reductase test and virtually all of us when you go into a physician's office, that is the dipstick that is being used, and it has a terrible sensitivity. And it is less than flipping a coin.

The other comment was that maybe the reason why the performance of the sensitivity data here is not as high as the Uriscreen is that the samples were not preselected for symptomatic patients.

DR. BEAVIS: Now, that was something that I was hoping I could follow up on if that is all right, because you mentioned that maybe the best utility of it is for the asymptomatic patients and directly culture the symptomatic patients.

But in the data that you presented, I didn't see that it was broken out in a way to see how this test works in the asymptomatic patient.

DR. MURRAY: But again we couldn't do that without getting informed consent, and if we got informed consent, then we would have had a very selective population of patients that we were analyzing.

And the feeling when this was discussed with the FDA was that they wanted to see the overall performance of the system with the types of patients that would have samples submitted to the clinical labs.

DR. BEAVIS: Thank you.

CHAIRMAN WILSON: Dr. Charache is next.

DR. CHARACHE: I have a couple of questions about the study and the study design. But I will say that with your dipsticks, the nitrate reductase, the directions are that you can't use that in the absence of the leukocyte esterase, but the two together give the predicted value, not either alone.

So that is just gratuitous. But I was looking -- I was very interested in the distribution of the species that were available, in part because the publication was overwhelmingly E. coli, and there wasn't much else there that we could look at.

Whereas, the data that you just presented now did give a nice display of other pathogens. But I did ask if there was any other data and the FDA sent me a long listing of the results.

It does seem to me that most of the E. coli came from one of your three sites. In other words, there was not an even distribution of the results.

The one side had a lot of contamination, and very little E. coli, maybe a half-a-dozen out of a couple of hundred. So I am wondering if you could tel us about the results by study site, and what the differences were between them.

And also how the contaminants were addressed. If you considered them culture negative, what happens if you look at that as a group to consider the false positive and false negative rate, and what did the contaminants do to your overall study.

And did it matter if their fecal contaminants are normal skin flora. I think that might also help understand how they fit together. So I am wondering about studies in different centers and also about the impact of the contaminants.

And my final question has to do with the fact that if you have 40 samples, and each one has to be tested for three minutes, that's a couple of hours at 30 degrees.

And I am wondering also about the effect of the first parts that are spending less time multiplying, rather than those that follow at the end of this two hour multiplication possibility.

DR. MURRAY: We will let John answer all of those. Actually, John does have the data where he has analyzed that.

MR. PLANT: We have looked at the breakdown of the false positives through the carousel to see whether there was more false positives at the end of the carousel rather than at the beginning, and in the second half of the carousel rather than int he first.

And there is no statistical difference between the two halves of the carousel. That was on occasions when there was a full carousel run of 40 samples.

DR. CHARACHE: I'm not concerned about the false positives, because you might just get a diagnosis of an E. coli, and you might get a positive because it is multiplied. But it would be below the detection limit had you done it earlier.

DR. MURRAY: The samples were -- to address that question, what they did was that they analyzed the 40 spots in the carousel with samples, and they repeated the testing of the same samples throughout the carousel, and then analyzed that to see what the sensitivity and specificity was.

And there was no difference in sensitivity or specificity for multiple samples, whether it was at the beginning of the carousel or at the end of the carousel run, because that was a concern.

DR. CHARACHE: Thank you.

CHAIRMAN WILSON: Mr. Travers, did you have a question?

MR. TRAVERS: Just as a follow-up on the question that you just made. Can I clarify whether you were worried about the bacteria going before we load them on to our system, or when they are loaded on to our system?

DR. CHARACHE: When they are loaded, and I think that may have been answered.

MR. TRAVERS: Yes, and basically we believe that the conditions that we put the samples in to promote the analyzing of the head-space are basically not very nice for bacteria.

DR. CHARACHE: And then I was wondering about the differences in results between the three study sites that you had, and my final question actually has to do with the volatiles that you are measuring. Are there species which these volatiles should not pick up that you might be concerned about?

MR. WHITE: Paul, do you want to answer that last one.

MR. TRAVERS: Yes. We've done several studies just looking at growing single species to see whether or not they produce these volatiles. And we have identified some yeasts which do not produce these volatiles.

They are not universal markers for every kind of infection that possibly could be present. It is a screening tool and it will pick out the ones that do produce these markers.

One of the markers is a general marker and is produced by lots of different organisms. The other marker is not. It is specific to one particular type of organism.

MR. WHITE: And John Plant will talk to you a little bit more about the differences between sites as well.

MR. PLANT: Just regarding that question, I don't think we have all of the data that you have asked for, but we looked at the sensitivity and specificity between each site and there was no statistical difference between sites for both sensitivity or specificity.

CHAIRMAN WILSON: Okay. Mr. Reynolds was next.

MR. REYNOLDS: Just to clarify something for me. Now, is it my understanding that if a patient is symptomatic the recommendation is that you don't screen this test, and that you go directly to culture?

MR. PLANT: Yes.

MR. REYNOLDS: That presents a major problem to me in the laboratory since most of the time I don't know what patients are symptomatic.

DR. MURRAY: It was a paradigm that was proposed, and actually Andy and I have discussed this, on how you would use a screening test in a laboratory. And you have a couple of options.

One is that you could screen every specimen that comes into the laboratory, and that is the way that this study was done. And the statistics were presented based on that.

The other is that you can have a physician make a decision whether they want to have a screening test performed, and presumably that would be done if there was a low index of suspicion of disease.

Or if they wanted to have a culture performed, or I guess you could have the contamination of doing a screening test and a culture. But presumably if you are going to do a screening test, and if the screening test is negative, you are going to stop there.

So that was a suggestion. But the way the data was presented was for all samples. Personally, I don't think you should have a symptomatic patient and ignore those symptoms.

I think a culture would be an appropriate or at least treatment would be an appropriate step. And I think it is misleading for us if we had just selected symptomatic patients and presented data on that, because the majority of the patients that we see are not infected.

So presumably most of those don't have symptoms, and those are the ones that we would like to screen and eliminate.

CHAIRMAN WILSON: Dr. Reller is next.

DR. RELLER: I have two questions. One, to follow up on this current thought, and the other was that the statement was that you wouldn't expect the test to work with boric acid preserved samples.

Let's come back to that. I would like to know what the chemical theoretical basis for that statement is given that the samples are put in additional compounds, and there is no growth.

In other words, you are not dependent upon growth of the organism for a positive test. Maybe we can handle this when we will come to the symptomatic and asymptomatic samples.

MR. WHITE: Can I turn back to Paul Travers, in terms of the boric acid.

DR. RELLER: So the theoretical basis for why boric acid would interfere, if it interferes.

MR. TRAVERS: Basically, the problem was when we did some initial studies was that the boric acid itself, the sensors on the sensor array responded to the boric acid in a way which was similar, or seemed to be similar to the way one of the market analytes responded.

So the fact that the boric acid could interfere with our assay at the marker chemicals, but in a subsequent study where we actually looked at boric acid at the levels that were used as a preservative, and we looked to see in a study whether boric acid interfered with the assay showed that it didn't interfere.

So in the end it was a precaution that I don't think we needed to take, but it was done anyway. But it was not so much that the boric acid is affecting the bacteria and changes the metabolites that are present, but rather is the fact that the boric acid itself could -- we were worried that it might be something that the sensors would respond to.

DR. RELLER: So that is a question to be answered later, and I don't mean later today, but I mean as in regards to the performance of this system with boric acid preserved samples.

MR. WHITE: I would say yes, but I think equally that the levels, in terms of what it is used for, in terms of transportation -- and back to the first question -- was that it wasn't an interfering substance at those levels.

Clearly, as we do for other studies, we made sure that was the case, and in terms of any interference data we ran, that was the conclusion of fact.

DR. RELLER: But no matter how this comes out, for the purpose of this discussion, there would have to be at this point an exclusion of samples with boric acid, because we have no data with the actual samples.

MR. WHITE: Yes.

DR. RELLER: Okay. Now, the second thing is, is it possible to go backwards on the slides to either the last or the penultimate slide that Dr. Murray showed with the algorithm, the proposed algorithm for use.

MR. WHITE: I think we have the technology.

DR. RELLER: Now, my question has to do with -- and in-part it has been answered, but if there is a high index of suspicion of a symptomatic patient, you are going to do the culture anyway or recommend it.

Now, let's go to the right side. If there is a low index of suspicion in an asymptomatic patient, I would like to dissect out what patients should be screened, whether it is with this or by culture, who are asymptomatic.

I think the only unequivocal patient who is asymptomatic, and who should be assessed, whether it is by screening or culture, are pregnant women with good antenatal care.

So my specific question is do we know how many of your patients whose urines came to the laboratory were done as part of screening; and I don't mean by OMA screening, but sent to the laboratory for the purposes of assessing presence or absence of bacteriuria as part of antenatal care.

DR. SCHAFFER: I will introduce myself. I am Anthony Schaffer, and I am a urologist from Northwestern University. I think what Pat Murray was alluding to is the fact that the majority of the samples were culture negative.

And some of those patients may have been symptomatic and had negative results, and he is assuming and I would agree since we do cultures frequently in our practice, that many patients have urine cultures who are not symptomatic, and I will give you examples of that. These are patients, for example, who are preoperative patients.

DR. RELLER: So that is another legitimate indication.

DR. SCHAFFER: Right.

DR. RELLER: Well, what I am saying is are there places before a procedure, like with pregnant women and diabetics,a nd so forth.

DR. SCHAFFER: Right. And spinal cord. There are a lot of reasons why you would want to know whether someone's urine was negative, and as a clinician that is what I really personally think this makes sense for.

And that is that if there is a high predictive value that the urine is negative, I don't have to do a culture. And in many patients that's what I want to know; that the asymptomatic patient, to be sure that the urine is negative.

DR. RELLER: There are recognized populations who would have a urine culture in the absence of symptoms, but they are not as nearly -- they do not constitute nearly as many patients as those who have cultures submitted to the laboratory for culture.

In other words, if a laboratory is getting a lot of specimens that shouldn't be sent in the first place, there is a lot of utility to doing something that would get rid of these and not bother with a culture.

On the other hand if there are legitimate patient populations who have no symptoms, but yet it is important to know before doing a urological procedure in the first trimester of pregnancy --

DR. SCHAFFER: Or diabetics, for example.

DR. RELLER: Well, you might educate me on the issue of screening diabetics, but apart from those who are not pregnant and who don't have a procedure that is planned --

DR. SCHAFFER: Children with reflux. There are a lot of these subsets that I would want to know had negative cultures who are not symptomatic.

A good example would be women who we see who have a history of recurring UTIs who are being followed and children who have urethral vessicle reflux, for example. So those would be the populations that one would want to make sure if you could had negative urines, and who might not yet have expressed symptoms.

DR. RELLER: Well, exactly. Do we have data on the performance of this approach to screening in those patients that we could come to agreement should be screened in the absence of symptoms?

To me it is a very important issue as to how -- and I am getting to my concern about the sensitivity of this test. The sensitivity of picking up the people that you really want to know whether they have it or they don't have it.

I can easily bury my questions about sensitivity if I am taking an HMO practice that -- you know, it is just one more cheap thing to send off to the laboratory, and most of them should not have been sent in the first place, and it looks like it performed pretty well.

But the pre-operative patient, the antenatal screening, et cetera, and from a clinical standpoint that we want to focus on, on how well does it perform that patient population.

And do we have any breakdown that would enable us to assess that from the thousand patients in round figures studies.

MR. SCHAFFER: My assessment is, since they didn't know the status of the patient, the answer is no.

MR. WHITE: I'll have John Plant respond.

MR. PLANT: We have to get the location of the clinic in the hospital within the data, and we can provide that to the FDA.

CHAIRMAN WILSON: I think we have time for about three more questions. Dr. Baron was first.

DR. BARON: I have a question about the test itself. Actually from the data that I have, I can figure out what you are measuring and you have four polymers. Is there a specific pattern that all four polymers give you for each of the two separate metabolites that you are measuring?

Or are two of them measuring something and two of them are measuring something else that you are paying no attention to? And why do you have 48 -- you know, 12 repeats of these four things?

How does that all work? Do four of them get used for one urine and then it moves on to the next four, and those recover?

MR. TRAVERS: No. To answer the first part of your question, there are four different polymer types, two of which respond to one of the marker analytes, and two which respond to the other.

I am not surprised that you were able to work out what the analytes are. The 48 sensors, we actually have on our sensor array 48 channels, where we can put down a sensor element and measure it.

We actually did a screening of the sensors that would be most useful for this particular application, and we didn't need any more than four. So we used the extra channels to basically put down replicates of each sensor type.

And basically what we analyze is the average of those 12 sensors, and so basically we are building redundancy into the system so that if one of the sensors starts to fail, there are 11 others that can still actually do the application.

DR. BARON: And it takes about three minutes for the head-space gas. Do you concentrate it down to such a fine stream that it takes three minutes to pass across the surface of the sensor? Why does it take three minutes?

MR. TRAVERS: If you actually look at the response profile, which is on the overhead at the moment, it is actually -- the three minutes is actually gas. For three minutes the nitrogen is passed across the sample, and initially it displaces the head-space that is there.

But then it is actually stripping out more volatiles, more of these analytes that we are interested in, and passing them across the sensor. The three minutes for the response profile kinetics for the market analytes are quite long, and so it doesn't actually reach equilibrium until about a minute-and-a-half into the response.

I think that is actually more of an issue when you are dealing with samples with relatively low concentration. So it is a compromise. We could have done the analysis in a shorter time, but we wouldn't have been as sensitive.

DR. BARON: Okay. So it takes three minutes and then you have a recovery period, and from what I heard from Pat, which I couldn't tell from the data, or I heard from somebody, it is 6 hours to run a 40 sample carousel through, 6 hours, start to finish?

MR. PLANT: No, it is six hours for the first sample result.

DR. BARON: For the first sample result.

MR. TRAVERS: And if it is a 40 carousel or 40 sample run, then you have a four carousel run, and it is about 15 or 16 hours.

DR. BARON: So it takes longer than a culture result almost, or just about the same amount of time than a culture. So why would I want to do this when I could have already cultured it and have my result the next day?

CHAIRMAN WILSON: I think that was a rhetorical question.

DR. BARON: You may certainly take it as a rhetorical question. I'm sorry.

MR. TRAVERS: I'm just not sure that I am the right person to answer the question. The question is really about why you would use the device.

DR. BARON: As a screening device, given the fact that you are going to have to save all those urines in the refrig, and then if this thing comes up positive on the screen, you are then going to go to those saved urines and culture them the next day.

And so in that time period you could have already cultured them, which is probably cheaper than what you are proposing to be done.

MR. GRINDROD: My name is David Grindrod, and I am the chief operating officer for Osmetech. The points you raised are very good ones, and I think there are two key ones that we would offer an explanation to.

First of all, the advantage that we have at the moment is that the device can be used by an unskilled operator. The result is positive or negative at the end.

DR. BARON: So you are saying that it is going to be a waved test?

MR. GRINDROD: So what I am saying is that you don't necessarily need to have the same level of skill that you would need to do a culture to be able to prepare it and get a result.

The second reason is that we are trying to provide a mechanism that avoids the need and the overhead for doing the culture in the first place. The timings that we have talked about today are very much aimed at developing a robust system of a novel technology we put through the FDA.

This is a first sample, a device that we have tried to provide something that has erred very much on the conservative side. As you have heard already, even on the sample time, that is where we believe that further work would enable us to be able to improve that.

So it is really about enabling technology and putting that through the FDA, and that is the reason why we are here today.

CHAIRMAN WILSON: Dr. Nolte.

DR. NOLTE: This point has been brought up several times already, but I am a little confused by the discussion about the appropriate statistic to look at for a screening test.

And I have heard some disagreement about whether sensitivity is really important here, but it is hard for me to understand how we can talk about this as a screening test and rejecting specimens for culture when we are going to miss 20 percent of the positives.

MR. ONDERDONK: I think in answering your question that the clinical study that was done here took all comers, and so we did not pre-screen anything. Were this system to be used as Dr. Murray's last slide showed, I am fairly confident that the sensitivity results would have been quite different.

In other words, we would have been taking a population where the expectation was that the samples would be negative, and I think you would find that both the sensitivity and the specificity would change as a result of that.

Alternatively, if we had done the very same thing that the Uriscreen did and take symptomatic patients and screen those, where the expectation is a higher number of positives, then I think you would have seen the sensitivity increase with this system.

We didn't do that in the clinical study that has been presented here. We took all-comers and we don't have a lot of patient information, including antibiotic use, which certainly can impact those numbers.

DR. NOLTE: But let me understand something. If you do that analysis, and the sensitivity still remains 80 percent in the asymptomatic patient population, do you still see value for this as a screening test?

MR. ONDERDONK: Well, I think that is up to the individual laboratories to decide quite frankly. I mean, I would certainly relish doing the study and looking at asymptomatic patients.

CHAIRMAN WILSON: And Dr. Nachamkin.

DR. NACHAMKIN: This is an analytical question. There is a little bit of a disconnect here between the assay, which to me should be a highly sensitive analytical assay, in terms of detecting small amounts of these volatile compounds.

And the low sensitivity in picking up 10 to the 5th organisms. When you did your kind of initial evaluation what was the lowest level of organisms that you could detect in spiked samples I guess is the question? And did that match what you found in your clinical trial?

MR. PLANT: We set our threshold up at one times 10 to the 5th, using a clinical trial.

DR. NACHAMKIN: Right. But that is not my question. In your pre-trial studies, you must have taken different urines with different concentrations or spiked normal urines with known concentrations of different organisms.

What is the actual minimum amount of the number of organisms that you can detect experimentally?

MR. PLANT: You have to remember that this is an indirect test. Although we have used clinical data to set our thresholds, there is not a direct link on the concentration of metabolites in clinical samples.

MR. TRAVERS: I'm not sure, but I think I can answer part of your question. When we did actually do single organism studies, where we basically tried to grow different levels of bugs in a urine sample and see what metabolites we got, when we actually did that, we couldn't detect below 10 to the 6th.

And we believe that part of that is the fact that how these things metabolized is dependent on the environment that they are in. I mean, we were putting them into a specimen jar with urine and leaving them with a temperature close to body temperature.

That is not the same as what is happening when they are in the bladder. Secondly, I am not a microbiologist and so I can't -- I am basically sort of reiterating what was being said in discussions with other people.

And it is apparently common that a single -- that for a particular bacteria you can have a single -- somebody help me. Clinical organisms are generally more virulent than single --

MR. PLANT: Than single strains?

MR. TRAVERS: Single strains, thank you. And that is one of the reasons why when we did -- I mean, this is going back a long way in the development of this instrument.

But when we did the initial work, we were very discouraged by it; and it is only when we went to clinical samples and we started looking at the results that we got from clinical samples that we realized that we actually could set the threshold at 10 to the 5.

DR. NACHAMKIN: So, I'm mistaken. I thought for some reason that this instrument would be highly sensitive in picking these things up, and in fact it is not as sensitive as you might want it to be analytically.

MR. TRAVERS: It is highly sensitive for the marker analytes, and it is the correlation between the concentration of those marker analytes and the level of infection.

We have configured our instrument to be able to detect at least 10 to the 5 level. We could do more work and change where we set our threshold, but there is work involved in doing that.

CHAIRMAN WILSON: There are a lot of questions, and we will do a few more. I think that Dr. Sanders is next.

DR. SANDERS: And my question had to do with the interfering substances. I didn't see pyridium urispas listed, and that is a common over-the counter preparation that can be taken if you have dysuria. Would that interfere with the test?

MR. PLANT: We don't have data on that.

CHAIRMAN WILSON: The next question will be from Dr. Janosky.

DR. JANOSKY: Yes. In looking through the data that you provided in the spread sheet, I see a fair number of system failures. What was the percentage of those system failures, and what was the cause typically of the system failures?

And then I also see data in there that would allow you to do subgroup analyses, and have you done any of those, or are those planned?

MR. PLANT: No, that's why we said --

DR. JANOSKY: So you haven't done any of them?

MR. PLANT: We haven't, but we can provide that information.

DR. JANOSKY: But what about the system failures?

MR. WHITE: We will go to slide number two and there is data as to that.

MR. GRINDROD: This deals with the bulk of the samples not analyzed. Two-thirds just under there are not device-related there, and they are basically categorized by samples not available, which I will cover in a moment, and an environmental temperature.

And these two separate events were -- one was where last Thanksgiving last year, and where they turned the air-conditioning off and the laboratory went out of range.

And I think it was one of the first falls of heavy snow in Boston in the beginning of December. So those are non-device related. We then have some device faults, and those are listed in the second point.

All of those particular faults were reserved, and we didn't see those particular problems reappear. If I can just move on to the next slide.

CHAIRMAN WILSON: Dr. Charache.

(Brief Pause.)

MR. GRINDROD: The other part of that answer on the samples no available was samples may have been collected, but the system was not available. We also have samples that were locked in, but samples no available, and that might be because they were lost or misdirected.

And we had 20 that were just straight database misallocation, and they came up as being not available for analysis, and we have four that were no culture results returned. So we passed them. I'm sorry, but there was a second part to your question, which I --

DR. JANOSKY: The second part was where I was asking whether you had any plan for the subgroup analyses. I know that a number of panel members had suggested that, and I was wondering if you had any plans to do so, or is that something that you are just hearing today?

MR. WHITE: That is just something that we are hearing today.

DR. CHARACHE: Hearing that, when you add that you need 10 to the 6th organisms to pick them up when you just inoculate that there were a lot of misses on that and that was originally discouraging; but that when you took clinical samples it worked.

It is highly reminiscent of some other studies, and I am thinking particularly of not only the leukocyte esterase, but the luciferase assay automation and so on, in which what was being measured was not the microbiology, but in fact the whole cell.

And my question here is with the volatiles that you are measuring are any of the metabolites consistent with human metabolism, whether it is leukocytes or bladder epithelium, or whatever, and have you looked for this? Are we looking at an inflammatory response or the microbiology?

DR. ONDERDONK: Some of these analytes certainly can be produced by eukaryotic cells. When we have looked at urine samples that do not have organisms and that we do not see these analytes.

So my assumption is that they are not produced in sufficient quantities for this system to detect them. But they certainly are absolutely unique to bacteria.

DR. CHARACHE: It may as I have mentioned -- and certainly this is what happened with the leukocyte esterases. It was -- well, not the esterases, but the luciferase assay. It turned out to be inflammatory cells that were causing the reaction.

And I am wondering if one had inflammation in the absence of bacterial cause whether you would -- whether it is chemical or whatever, whether you would get a false positive?

I think that this might be important in terms of what it is that you are actually measuring, and therefore where you would expect your false positives, and particularly your false negatives?

And since we are talking about this being used to screen asymptomatics, you may actually increase your false negatives if it is not associated with an inflammatory reaction.

DR. ONDERDONK: That's an excellent question, and I don't have any data to support or refute anything you said.

CHAIRMAN WILSON: Okay. One final question. Dr. Durack.

DR. DURACK: Does the polymer sensing characteristic appear right over time, or does it need to be regenerated after use? I guess it is to do with the lifetime of the device, and does it need to be stripped after it has done a sensing round?

MR. TRAVERS: The polymers -- one of the things that we were conscious of when we were designing the instrument is that sensory systems are subject to drift and that can be either down to effects in temperature and humidity, or for aging of a sensor if sensor characteristics would change over time.

So what we actually did was to -- we set up our protocol for sampling in such a way that we would detect if a sensor response starts to drift, and would recalibrate if necessary.

So that is why we have a system check carried out every day, which basically checks if the response is still the same as it was during calibration. If it is, then you can carry on and process samples.

If it fails a system check, then you recalibrate the system, and so effectively you are recharacterizing your classification thresholds to track any changes that might occur in a sensory response over time.

DR. DURACK: And is that daily or every run, or what?

MR. TRAVERS: Over the course of the performance trial, which was carried out over three sites and several months -- and this is just off the top of my head -- we had to recalibrate -- and this is three systems, but we had to calibrate twice, recalibrate twice.

DR. DURACK: But it was the same sense for the three months?

MR. TRAVERS: Yes.

DR. DURACK: Thank you.

MR. TRAVERS: It was on one site that we had to replace the sensor in the middle of the trial, but the other two sites we used the same sensor.

CHAIRMAN WILSON: Okay. I would like to thank the sponsor for their presentation, and at this time I would like to move to the FDA presentation.

MR. WHITE: Thank you very much indeed.

MS. HEYLINER: Good afternoon, Panel Members. The sponsor has presented the facts of the OMA-UTI and we are in agreement. I just want to remind you that this submission is being reviewed as a 510(k).

Usually we bring to the panel PMAs to demonstrate safety and effectiveness. With a 510(k), we try to demonstrate substantial equivalence to other legally declared marketed devices or predicate devices.

The topics that I intend to touch very briefly on is just on the background and a little bit about the technology, and the study results, and the discrepant results and the conclusion.

The FDA has cleared a variety of screening devices for detection of negative urine specimens that do not require further analysis for organism identification.

Some of the methods currently available vary widely in levels of automation, technologies, and means of interpretative criteria. Such methods, as the measurement of biolumiesence, electrical impedance, automated urine sediment staining, catalase testing, and urinalysis by dipsticks.

But the quantitative urine culture remains the standard practice. The OMA-UTI differs in technology from all other cleared devices. As the sponsor explained, volatile compounds produced by bacteria in the headspace of the urine sample tube, these are the compounds that are being detected and they are detected by an array of specific conducting polymer gas sensors.

The samples are then classified as positive or negative, using Principal Components Analysis. Now, this submission had a lot of technological considerations for us because it was a new technology.

So in our review, we considered some of the parameters that could influence performance of the OMA-UTI, and these were things like determination of the discrimination threshold, and the constant concentration vector of the principal components, the stability of the OMA-UTI detector, since sensor drifting is known to affect performance of array sensors.

And also we looked at the nature of the sensor material, because gas can sometimes interfere. Now, if we looked at the study done by Osmetech, there were 1,038 samples that were analyzed, and significant bacteriuria was defined as over 10 to 5 colony forming units per Ml.

Well, you have seen this chart before, and I won't go into the details other than to point out that there were 151 samples that you could probably call as positive, and 28 that could be regarded as false negatives.

As we mentioned before -- the next slide, please -- the OMA-UTI have the following performance characteristics relative to standard culture: the sensitivity of 81 percent; and the specificity of 83 percent; a positive predictive value of 44 percent; and a negative predictive value of 96 percent.

There was no patient clinical chart to review in order to determine what the clinical significance of these discrepant cases might be, and so we were not aware of whether the patient had fever, or whether they had a blood culture, or urinalysis.

And I think the manufacturer explained the reasons for that.

The predictive value was 44 percent, and it is generally conceded that urine screening methods have a low positive predictive value, and they are unreliable for UTI diagnosis.

The negative predictive value was 96 percent. A screening method with a high negative predictive value usually has high utility in identifying non-infected urine specimens and excluding them from further examination.

Now, let's look at the false positive results. There were 141 false positive results, and in fact the sponsor attributed them to the fact that there might be a higher proportion of negative samples in the study population, because 83 percent of the population was in fact negative samples.

The OMA-UTI might be measuring metabolites that are produced by a bacteria before reduction of numbers by antibiotic treatment. The might be organisms producing higher levels of metabolites, but whose standard culture results might be below the predefined threshold.

Or there might be samples with metabolite concentrations falling at the detection threshold, with a 50 percent chance of being reported as either negative or positive.

Let's look at the 28 false negative results. What could they be attributed to? Probably volatiles from some species may not be detectable by the present sensor array system.

And in looking at the data it was noted that there was a low sensitivity with enterococcus and yeast, and E. coli perhaps. While bacteria may be lost by absorption on to urinary cells, or protein, or by participation between specimen collection and analysis.

Volatile substances in the urine might saturate the sensor detectors and block the response to bacterial compounds by competitive inhibition. So the OMA-UTI is intended for use by clinical lab health care professionals as an aid in the detection of bacteria associated with urinary tract infection.

It indirectly measures bacterial infection by a semi-quantitative analysis of volatile compounds released into the head-space above a urine sample. But compared to the predicate Uriscreen, which actually just detects catalases, we have with this device new technological characteristics to consider.

And that is the reason why we are here, because we would like to have your input as to how best we could probably adapt this new technology to diagnosis of urine in the clinical lab.

And these are the members of the review team who worked on this submission. They are Ellen Chen, from the Office of Science and Technology, and she is a polymer chemist; John Dawson, our biostatistician; Jean Fourcroy, Medical Officer, and myself.

CHAIRMAN WILSON: Thank you. Do any of the panel members have questions of the FDA? Dr. Nolte.

DR. NOLTE: At the risk of sounding like a broken record, the sensitivity was missing from your criteria for an acceptable urine screening device, and I am curious why it falls off your table as well?

MS. HEYLINER: Well, you know, I recognize that sensitivity is important, but I think that perhaps because I am thinking of substantial equivalence, I am looking to see if this device is in fact comparable to other legally declared market devices.

And I am looking at this device as a screening device, and so I am more concerned with its negative predictive value. I mean, your point is well taken about sensitivity. It probably is one of our concerns, but I guess I didn't give it a lot of importance here.

CHAIRMAN WILSON: Dr. Gutman.

DR. GUTMAN: Yes. We are bound by history and so we can't, whether we like it or not, acquire a lot of different performance in a new device. So I suspect that there is a wide range of devices besides the one that the sponsor has shared with you, which probably with performance is not much different than this.

So we are actually not asking you to help us here. I think we are -- that our law allows us to be substantially equivalent, and it doesn't have to be any better and it shouldn't be much worse.

And we may have actually deliberately or inadvertently misled the company into the data presentation that they put here, because that is how we think that will actually be generated.

That having sort of an uncontrolled data set that came in that isn't screened for asymptomatic and symptomatic strikes us as probably real world, and what we really want to do when we label this product is not have allusions about how it might perform.

So if you think that is bad, you can certainly let us know, or if you think that some subset of analysis is appropriate, that is probably a good idea.

But we probably negotiated with the company and said give us something that is real and that comes from real labs, and not something that is highly contrived and likely not to reflect the product in use. And again any advice that you may have will be welcomed.

CHAIRMAN WILSON: Dr. Charache, you had your hand up next.

DR. CHARACHE: Yeah, I did. I think I was on a similar track. I think that the problem is probably the first horse out of the barn, because obviously we don't do a urine -- we don't take a urine specimen to prove that it is negative.

We take it because we want to rule out infection, and this device misses one in five, and that is a lot of misses. But it parallels the same experience with a lot of other tests that are already out there on the market.

But what I would like to question because of the change in technology, is if there are populations that are negative -- and I think particularly the issue that we raised just a few minutes before, that if this in fact requires inflammation, and perhaps polymers, in order to get a positive which has rapid metabolic activity and perhaps of the same volatiles, I think it would be helpful to screen and get some data on patients who have aplasia, and perhaps the oncology population.

We have done this kind of thing with some other tests. We looked at outpatients, versus inpatients, and patients who had turbidity versus no turbidity, and this kind of thing, because a lot of the turbidity of course is cells.

So I think that it might be helpful to know where it should not be used as a screening procedure because of its technology.

MS. HEYLINER: Certainly. We are still working with the company on this device, and it is still under active review and so your suggestion is well taken.

CHAIRMAN WILSON: Dr. Nachamkin.

DR. NACHAMKIN: So I'm a little confused now, in terms of the indications for this device, and I will tell you why. Because in the package insert, in the revised package insert, under interpretation of results, it says that a positive result is indicative of UTI and correlates the production of volatile compounds from greater than 10 to the 5th CFUs of either single colonies or from mixed colonies containing at least one predominant organism greater than a hundred-thousand CFUs per Ml.

The predictive value of this test is only 44 percent, and I thought the test was being proposed as a screening device for laboratories to decide culture or not cultural and not to give clinicians an answer that patients got bacteriuria or not.

So there seems to be some -- what it says here is not what we have been hearing during these conversations, and obviously there is a labeling concern, but I guess the question is that since it was written like this does the company feel that you could report this as a screening device that is positive to clinicians?

MR. GRINDROD: We believe that the screen is a utility and not that it reports the positive results, but that reports those samples that are negative.

DR. NACHAMKIN: Then may I ask why do you have that actually in the package insert?

MR. GRINDROD: I think that is a very good question.

CHAIRMAN WILSON: Dr. Baron.

DR. BARON: To go back to Marian's question about the predicate device. Let me ask a question about Uriscreen. I was looking at catalase, and one would expect a Group B strep infection not to have a whole lot of catalase because Group B strep doesn't make catalase.

I am not sure about the PMN catalase. Is that -- Pat, were you alluding to the fact that your screen positivity also is positive in patients with a lot of PMNs, but not bacteria at all in the Uriscreen?

DR. CHARACHE: Yes, possibly. Maybe it is chlamydia. I don't know.

DR. BARON: Okay. So a positive Uriscreen could be due to catalase caused by human cells or bacterial cells. So that you would not miss necessarily those catalase negative bugs, like Enterococcus and Group B strep.

But this device would miss -- because I think it is fair to say that those metabolites are less likely to be produced at the level that would indicate greater than 10 to the 5th bugs, even if they were being made by some PMNs, or else we would not have seen so many negatives in this.

MS. HEYLINER: Yes, I agree. The Uriscreen is actually for the detection of catalase in white blood cells or bacteria if I remember correctly. It was quite a few years ago, but I believe that indeed was the intended use.

And because this test is detecting the volatiles, you will in fact miss probably the Enterococcus, the E. coli, and the yeast, but you probably would have picked up if that sample had been done by Uriscreen.

DR. BARON: So I think there are really different technologies.

MS. HEYLINER: Yes, but when we chose the predicate, we actually are looking for intended use. When we compare one thing to the other, we really look to see if the intended use is similar, even though the technology might be different.

CHAIRMAN WILSON: Dr. Reller.

DR. RELLER: I wonder about the screening device's inordinate emphasis on negative predictive value, as opposed to sensitivity. There are patient populations who should be screened heretofore by culture, and I don't know quite honestly whether some of the other approved devices for screening actually exclude these patients.

But, for example, if one accepts that what you are picking up in pregnant women, screening for bacteriuria, are those persons who have asymptomatic bacteriuria that cast years ago showed about one percent or 1-1/2 percent per decade of life.

So if you had elderly people, maybe 10 percent asymptomatic bacteriuria of no clinical importance, unless you are being instrumented, et cetera.

But let's say it is 3 to 5 percent in the population of pregnant women. Well, right off the bat before you do anything, you have a negative predictive value if you put the sample down the drain of 95 percent.

And what I want to know is in those patients whether I am able to pick up those individuals who left untreated will get into complications at a far higher percentage. I mean, on the order of the published figures of 30, 40, or 50 percent, if untreated will come to a symptomatic infection, with the consequences to premature delivery or many things.

I mean, it is good to detect, and find, and treat. So there are -- either we have the data that it is good enough for those asymptomatic patients, or there is an exclusion that there are no data and it should not be used for that purpose.

And when you start not having the information on specific groups of asymptomatics, which I don't think we do have, and then we have a more general recommendation that if you are symptomatic that you should do the culture anyway, then predicate devices that are already on the market notwithstanding, I think some of the same comments could be made for them. I think we have problems.

MS. HEYLINER: The data that was presented was the data that we got from the sponsor. However, as I said, the 510(k) is still being actively reviewed, and one of the questions that we asked the sponsor, because I think we did feel the same way like you do, that there were other groups that probably should have been addressed, such as diabetics, and pregnant people, children, you know.

And the sponsor I think -- and I don't want to speak for the sponsor, but I think the sponsor intends to address that in their labeling perhaps if we can't come up with that data. So that there still might be a use for the device.

CHAIRMAN WILSON: All right. Are there any other questions for the FDA? Dr. Beavis.

DR. BEAVIS: I wanted to second what Dr. Reller was saying. Given that the predicate advice -- you know, the Uriscreen, the sensitivity for that is 95 percent, compared with 81 percent for the OMA, and I think to measure the sensitivity is a lot more relevant than the measure of negative predictive values, especially since there are only 13 or 14 percent positive cultures in the study, I believe.

And the other thing, too, is that I know that we spent a lot of time, and I am interested, too, on whether you can differentiate between the asymptomatic and the symptomatic patients, and whether their specimens should be screened or not screened.

But to me the bottom line still is that it is missing 19 percent from symptomatic or asymptomatic people.

CHAIRMAN WILSON: Okay. There is time for one more question. Dr. Nolte.

DR. NOLTE: No.

CHAIRMAN WILSON: Okay. Thank you. At this time, I would like to open the meeting to the open public hearing portion. Is there any members in the audience who would like to make a statement.

(No audible response.)

CHAIRMAN WILSON: If not, then the public hearing is closed, and at this point I would like to move on then to the open committee discussion. I have already asked our primary reviewer if she would like to make any initial comments, and she has said no, and therefore, we would like to begin with the questions.

So if we could have the first question, please.

The first question posed to the panel is to please comment on the adequacy of the data presented to support the use of the device as an aid in the detection of bacteria associated with UTI. Do we have any comments from the members of the panel?

DR. DURACK: Well, to start the discussion, I think there is again some lack of clarity here. The package insert that is proposed uses this wording, "aid in the detection of bacteria," but the presentation seems to have emphasized aid in the exclusion of bacteria. So I think we have got to resolve that before we can really go forward.

CHAIRMAN WILSON: Dr. Gutman, would you like to clarify that for us.

DR. GUTMAN: Well, I think the sponsor has clarified the intent, and so the labeling needs to reflect it. So given the fact that I think the intent of the sponsor is to rule out infection rather than to establish the present infection, the question that you need to address is whether this is the right data, and whether you want to ask for other data.

And although it would be difficult for us, if you wish to suggest other performance parameters,

you can put any of those on the table and we will do the best that we can.

CHAIRMAN WILSON: Suggestions or comments? Dr. Nolte.

DR. NOLTE: Do we have -- I know that we have said the word screen an awful lot, but all I keep seeing in terms of the printed material is an aid, and are we to --

DR. GUTMAN: Don't worry about that. We will surely fix that. We will fix that, and we will refocus it to be what the sponsor is trying to sell here, which is I think a test to rule out the presence of a need to culture requirement.

CHAIRMAN WILSON: Additional comments? Dr. Charache.

DR. CHARACHE: Well, I have just been quickly also looking at the printout, and I have seen a number just going through that were E. coli grew, or klebsiella, or pseudomonas, in which there was a negative result. They were falsely negative.

And at least four of them come from organ transplantation. I really think we need to know more about the patient populations and we know in whom it would not work. And we might get a better understanding of why it works.

The assay that I was referring to before was one that I really feel strongly about, and was the only time that I ever returned money because I wouldn't continue the study.

But that was one in which the detection system was detecting the ATP, the luciferase assay detecting ATP, from 10 to the 6th bacterial, or 10 to the 5th bacterial.

And it turned out that one leukocyte had as much ATP as 10 to the 6 pseudomonas. And when we corrected for that, we knew what it was measuring. A hospital in Boston took the money.

CHAIRMAN WILSON: Dr. Baron.

DR. BARON: As Dr. Charache has mentioned much earlier, there is a big discrepancy or a big difference between the kinds of patients and the kinds of results that are reported out by the three different groups that evaluated urines for this study.

One of them had a lot of contaminants, and one of them had a whole lot of E. colis. So I think rather than have it in one giant chart like this, I would like to see the data broken down by patient gender, patient age, type of patient, what kind of ward the patient came from.

And then whether there were contaminants for positives versus negatives, and what they actually grew. I would like to see -- you know, this is very hard for me to look at line by line, and so I would really like to see those data broken out in a different way.

CHAIRMAN WILSON: Okay. Any other comments on the first question? Dr. Nachamkin.

DR. NACHAMKIN: Dr. Murray mentioned before that one of the reasons why you didn't stratify patients by asymptomatic versus symptomatic is that you needed to get informed consent, and that decreased the complexity of the study.

It is not clear to me that actually you need to have informed consent on the de-identified data, and --

DR. BARON: In my hospital you would.

DR. NACHAMKIN: I am not sure that IRB would require informed consent for that specific piece of information. Pat, did you actually talk to your IRB about this?

DR. MURRAY: What is the question?

DR. NACHAMKIN: The question is that -- well, one of the issues is knowing the performance of the test in asymptomatic versus symptomatic.

DR. MURRAY: We would have to have a chart reviewed to determine that.

DR. NACHAMKIN: And you would have to have informed consent to do that?

DR. MURRAY: Yes. I am sure that Hopkins would, too.

DR. CHARACHE: Hopkins' requirements are in flux.

DR. MURRAY: We did record data that was available when the patients came in to -- when the specimens came in to the laboratories, and so we do have hospital location.

So we can go back and reexamine that data, but we can't tell if the patients were on antibiotics because obviously that data is not accurate than what is on the requisition.

And certainly it is not indicated that they are symptomatic or not, and so we would not have been able to get that data without informed consent.

DR. NACHAMKIN: How about comparing it with the UAs on these patients?

DR. MURRAY: Not all of the specimens had Uas, and that was not done in the patients, and the patients did not have that.

CHAIRMAN WILSON: Dr. Charache.

DR. CHARACHE: If you were going to look at another subset of patients, an easy way to screen for antibiotics is just to make a lawn of the coag-negative staph, and you dip a filter paper disk in the urine and put it on the lawn, and you can put a lot of patients on one plate.

DR. MURRAY: Do you have sensitive ones? We have patient isolates.

DR. CHARACHE: No, these are not patient isolates, but we have used that technique to correct for antibiotics in other studies.

DR. MURRAY: That's a good suggestion, but obviously it wasn't done in this study.

CHAIRMAN WILSON: Okay. Any further comments? If not, then I would like to have the second question. Okay. The results of the UTI when compared to standard cultures showed a high number of false positive results.

Given the confounding factors such as reduction and bacterial numbers due to antibiotic use, or production of high levels of metabolites with some bacteria, are there any other comparative methods that may be more appropriate? Any comments from the panel on that issue? Dr. Baron.

DR. BARON: I frankly don't have a big problem with false positives if we are looking at a screening test.

DR. NACHAMKIN: I second that if that is the indication.

CHAIRMAN WILSON: Okay. All right. Can we have the third question then. The third question states the detection thresholds are the only UTI that has been set to detect levels of volatile metabolites found in specimens with bacterial counts greater than or equal to 1 times 10 to the 5th CFU per Ml for either single colonies or mixed colonies containing at least one predominant organism at the same concentration.

Should the package insert address bacterial counts below 1 times 10 to the 5th, and if so, how. Dr. Charache.

DR. CHARACHE: I would also like to address the contaminants, plus one predominant organism. When you have normal fecal flora, you certainly can have a predominance of E. coli and an irrelevant culture.

So I think that is a bit problematic, and I think we should analyze that group separately. In terms of less than 10 to the 5th, the studies by Kunin and Kass, one in school children and one in hospitalized patients, both showed about or between 15 and 25 percent, more being around 20 to 25 percent, of 10 to the 4th for significant urinary tract infections.

That is certainly true of yeast. If you get more than 10 to the 4th, they have taken it from the bag and not from the patient. But I think also that what you are supposed to do under those circumstances is to repeat the culture, and if you have two 10 to the 4ths, that equals 10 to the 5th, and it equals a urinary tract infection.

So I do think that it is a degradation of information when you limit it to 10 to the 5th. Now, I think that this has to be put into perspective with other assays that are out there, in terms of regulatory need.

CHAIRMAN WILSON: Dr. Baron, did you have a comment?

DR. BARON: It a routine clinical laboratory many patients' urines are considered positive at 10 to the 4th. Patients who are catheterized in the hospital, if you follow them day after day, their numbers are low to begin with, but they still legal up, and if they are pure, they repeat.

And as Dr. Charache has just mentioned, we consider them to be positive. So I think the 10 to the 5th cutoff would serve very well for those asymptomatic patients that Dr. Reller has been describing, where the threshold for a positive bacteria would be 10 to the 5th.

And in that patient group I am extremely concerned that we don't have the data here to see if 10 to the 5th sensitivity holds up in that group where 10 to the 5th would be the appropriate threshold, because I think in a hospitalized patient, or a symptomatic patient, 10 to the 5th is not the appropriate threshold.

CHAIRMAN WILSON: Okay. Any additional comments? Dr. Charache.

DR. CHARACHE: Yes. I wonder if we could exclude certain patients. In other words, have as a requirement that it be a clean catch, and that it not be a super pubic specimen.

CHAIRMAN WILSON: Okay. And can we have the fourth question. This is a request that we please comment on the warnings, limitations, and precautions in the labeling. Does anyone have any comments on that issue? I think we have discussed that to some extent already.

DR. BARON: Yes, I have one that was not brought up. Somewhere in the product insert, and elsewhere, it said 12 hours, and I think it said that the sample should be tested within 12 hours.

It says 24 in one place and 12 somewhere else. So I just have this problem with the discrepancy in the number of hours, and I have to go look through my book and find out where it said 12 on the revised product inserts.

Here it is. It says, "Tests within 24 hours, store up to 12 hours." That is where the discrepancy is.

CHAIRMAN WILSON: All right. Any other comments on the labeling? Dr. Nachamkin.

DR. NACHAMKIN: I don't remember seeing this, but you want to make sure that there is a specific comment in there that says that the urines should be refrigerated during the test procedure prior to deciding on whether to culture or not.

I didn't see an explicit statement about that, even though in the lab we know that urines should be refrigerated until they culture, and I think it should be stated outrightly within the package insert.

CHAIRMAN WILSON: Dr. Baron.

DR. BARON: On page G-2 of the package insert under warnings, it says, "A negative test result does not in itself rule out significant bacteria. There are occasionally UTIs caused by organisms that may not be correctly identified as positive." But actually there are negative results caused by organisms that should be positive, like E. coli, pseudomonas, et cetera.

CHAIRMAN WILSON: Okay. Thank you. Dr. Durack.

DR. DURACK: With regard to the wording on the indications or on the intended use, Dr. Gutman has told us that will be revised, but I think we have to come back to an issue which very much affects what I think the sponsor would want.

The sponsor presumably would like to have the indication to be exclusion of infection in asymptomatic patients. I would think they would like that, but we don't have the data for that group.

And Marian pointed out that the positivity rate was only 17 percent, but in the asymptomatic group it may be much less than that, much less than that, but particularly in subgroups.

So I think that this is just an area that has to be dealt with, but we are looking at a device where one of the primary applications we may not have the data for.

CHAIRMAN WILSON: Dr. Nolte.

DR. NOLTE: I am not sure that we don't. I mean, we are thinking about this in terms of the sensitivity issue again, and the negative predictive value is going to change if we segregate out the symptomatic from the asymptomatic patients.

But I am not sure -- well, what we have here is a test for 10 to the 5 organisms per Ml, and I don't see how that is going to -- how the sensitivity is going to be influenced much by the pretest probability.

DR. DURACK: It is possibly not -- what if the host has an effect, which could be particularly applicable in asymptomatic patients. I just raise it because the intended use is going to be very important.

DR. NOLTE: I'm with you a hundred percent.

CHAIRMAN WILSON: Dr. Nachamkin.

DR. NACHAMKIN: I think the -- if I understand your question correctly, did you state that you think that the sensitivity is going to be stable over different populations?

DR. NOLTE: I'm guessing since from the data that we saw that there is a relatively low positivity -- I mean, what is it, 13 or 17 percent of the patients were positive.

And I think I heard Dr. Murray say something about that he suspected that most of the patients were probably asymptomatic, and just thinking about this as a test for bacteriuria, that is really what it purports to be.

There is no reason in my mind to think that the sensitivity is going to change substantially should we include just asymptomatic patients. But maybe I have got it wrong.

CHAIRMAN WILSON: Dr. Charache.

MS. POOLE: I think this comes back to what we said, but in terms of specific wording, under warnings on page G-2, warning number two, the last sentence there -- if clinical signs and symptoms are suggestive of a UTI -- for example, increased frequency, dysuria, and urgency, retest with a new sample or an alternative method is recommended.

I think that really should be culture is recommended, because you are going to waste another 24 hours with a patient with a UTI. Repeating it, you may get the same answer.

CHAIRMAN WILSON: That's a good point. Okay. Dr. Reller.

DR. RELLER: Two things. One is my comments about asymptomatic patients has already been made in lacking the data. On the symptomatic patients, unless one accepts that it is not a consideration there at all, a substantial number of symptomatic patients, the threshold for detection of important bacteriuria as has been mentioned is lower.

From the theoretical basis for this test, we have every reason to expect that the sensitivity as a screening technique in patients with single organisms -- 10 to the 4, for example, who are symptomatic -- may even be less.

So if the sensitivity is 81 percent all-comers, at 17 percent overall positivity, what is going to happen with symptomatic patients with lower numbers, and I don't want to miss the patients who have bacteriuria, even if it is only five percent of them in some of these populations already mentioned.

The second comment is actually a request or a query to our statistical consultants, and that has to do with given the numbers of samples in this study, if we look at the sensitivity with the Uriscreen in the sheet provided, and the sensitivity in the OMA approach, what are the confidence intervals, and are those different, or do they overlap when one of the queries is of substantial comparability?

CHAIRMAN WILSON: Dr. Janosky.

DR. JANOSKY: I don't know if the FDA statistician is still here. I didn't calculate confidence intervals. Did the sponsor calculate confidence intervals?

MR. GRINDROD: We have the confidence intervals for our device, and I think in one of our slides, which I can put back, but we don't have them for the predicates.

CHAIRMAN WILSON: Okay. Any further comments?

DR. JANOSKY: You can somewhat -- let me just -- well, if you look at the confidence intervals that are provided in the panel packet, and you look at the predicate device values, you can try to match up those values with the confidence intervals to see if they overlap or not.

The predicate devices do not have confidence intervals on this slide, and so that is one way you can answer the question that you are asking. And it looks like for some of them that they are outside of the confidence intervals.

So if you look at the predicate devices, they are outside of the confidence intervals provided for the device that we are looking at today.

DR. RELLER: Which page? What are those confidence intervals for this OMA?

DR. JANOSKY: I am looking at -- it looks like this is the FDA presentation to us today. It says, "Performance Characteristics," at the top. I think it was one of your slides, yes.

DR. GUTMAN: Well, John is obviously gone, but it certainly is a question that we could ask him, and we can certainly query. We should have access to the data in the previous submission.

And so if they weren't calculated, it should be possible to go back in to calculate them.

So we can't answer it now, but it is answerable, I think.

DR. RELLER: What we have here is sensitivity, 81 percent, confidence interval, 74 to 87. And I don't know whether that 95 in the predicate is -- I mean, I don't know whether that overlaps or not. Now, not being a statistician -- I mean, 81 and 95 don't seem the same to me.

DR. JANOSKY: Chances are that with a sample size that it is much smaller for the Uriscreen, and so the confidence interval is going to be wider in that respect. But without having the actual values here, it is only a guess as to whether they would or not.

CHAIRMAN WILSON: Okay. Any additional comments? Dr. Gutman, in terms of the final recommendations, I think we would have to ask if we have given you the information that you need to do it.

DR. GUTMAN: Well, you have given us a lot of food for thought; and again, how much latitude we have here in responding to all of this is something that we will explore. We will go back and look at he predicates and see.

I do sense a certain concern about whether the dataset that we are looking at matches the claim and also about the sensitivity. And so our challenge is to go back and see how many -- you know, see what we can do to address those in either the existing data or we want to negotiate with the company to give us more data.

And then what we can address in the review process, and what we can address in the labeling. I can tell you that we have a long history of products that have done this, and my guess is that some of them will not have fantastic performance, and that it would be legally challenging for us.

And not legal challenging perhaps to get better characterization of the data, and to get honest labeling. It might be legally challenging not to allow better characterized and better labeled product to be on the market. But you have given us a lot of food for thought and so thank you.

CHAIRMAN WILSON: Okay. Are there any final recommendations that the members of the panel would like to make? Dr. Baron.

DR. BARON: On the proposed labeling that the company responded to the FDA's queries, it says only appropriate trained clinical laboratory health care professionals should operate the equipment.

And based on what I have read about the operation, and particularly the calibration, I would agree with that. But when I asked you the question about what would be the advantage of it, you answered me that it would be able to be used by less trained personnel.

So I think you probably need to sort that out a bit and figure out just who it is that you want to be doing this. I asked you would it be a waived test if you were going to use it in the setting of a screen at a nursery school, or an out-of-the-lab clinic, then you would want to make it a simpler instrument that would work better, and you wouldn't have to have that kind of labeling.

CHAIRMAN WILSON: Dr. Reller.

DR. RELLER: Fortunately, I am not in the difficult position that Dr. Gutman is in and colleagues at the FDA. But from a clinical microbiology laboratory public health perspective, surely there must be some innovative way to keep the bar high scientifically.

I wish there were a way to address the things that are already available that might never pass muster if they were looked at currently. We need -- patients are not simpler than they used to be. They are more complex, and the laboratory is hamstrung in the amount of information needed to appropriately test, and apply testing, and give a clinically useful result. So that some of the very populations from which we receive specimens, there may be approved products that are sadly wanting in practice.

And I don't know how this issue can be addressed, but I think it is an important one to be considered for the agency for the future. Maybe for what it is worth, you know, an advisory committee's recommendation, with the examples of things that could.

And the agency has its mandate from legislative action, and there could be additional legislative action that would give the FDA the wherewithal to do its job serving the public even better in my view.

CHAIRMAN WILSON: Dr. Carroll.

DR. CARROLL: I would just like to make one additional comment. This is a new technology, even though we are comparing it to existing predicate devices.

And I just want to come back to a couple of the points that some of the other panelists made with respect to additional data on interfering substances like pyridium, as well as looking at other sources of volatile compounds other than microorganisms. So I do want to come back to that issue as well.

CHAIRMAN WILSON: Okay. Any further comments? Okay. I would like to thank the members of the panel for this discussion, and thank the FDA for their presentation.

And I would particularly like to thank the sponsor for all the work that they did, and for all of the members of their team who made the trip here. And if there is no further comments, I would like to adjourn the meeting.

(Whereupon, at 6:20 p.m., the meeting was concluded.)