Tests, Detection and Diagnosis

 

 

Abraham, A., V. Sivanandan, D.A. Halvorson, and J.A. Newman (1986). Standardization of enzyme-linked immunosorbent assay for avian influenza virus antibodies in turkeys. American Journal of Veterinary Research 47(3): 561-6.  ISSN: 0002-9645.

            NAL Call Number:  41.8 Am3A

            Abstract:  The signal-to-noise ratio was useful in determining the optimal dilution of rabbit anti-turkey conjugate. Optimum dilution for rabbit anti-turkey conjugate to be used in the enzyme-linked immunosorbent assay (ELISA) was 1:1,000. The avian influenza virus antigen concentration was 128 hemagglutinating units (0.3 microgram of protein) per well, as determined by checkerboard titration. Bovine serum albumin fraction V increased nonspecific binding of conjugate and was not used to coat the plates in subsequent tests. Using ELISA, nonspecific binding to avian influenza virus-coated plates were not found with antibodies to Newcastle disease virus, infectious bursal disease, Salmonella, or Escherichia coli. Chromogens o-phenenediamine, and 2,2'-azino-di-(3-ethyl-benz-thiazoline sulfonic acid) were almost equal in sensitivity for detecting released oxygen from the H2O2. The substrate plate was more sensitive than was the polystyrene plate. Dual wavelength was reliable in reading ELISA results.

            Descriptors:  antibodies, viral analysis, fowl plague immunology, influenza A virus avian immunology, enzyme linked immunosorbent assay, hemagglutination inhibition tests, turkeys.

Abraham, A., V. Sivanandan, J.A. Newman, and S.K. Maheswaran (1984). Rapid purification of avian influenza virus for use in enzyme-linked immunosorbent assay. American Journal of Veterinary Research 45(5): 959-62.  ISSN: 0002-9645.

            NAL Call Number:  41.8 Am3A

            Abstract:  A rapid and easy purification method was developed to obtain avian influenza antigen for use in immunochemical assays. This was achieved by rapid concentration of virus from infective allantoic fluid, using 8% (w/v) polyethylene glycol 8000, and later, by purification on gel-permeation chromatography through controlled-pore glass beads. Rabbit anti-turkey globulins were made specific for turkey globulins, using affinity chromatography, conjugated to horseradish peroxidase and used in enzyme-linked immunosorbent assay. A significant increase in specificity and sensitivity of the enzyme-linked immunosorbent assay was observed when purified antigen was used in place of a crude antigen preparation. This purified antigen eliminated the false-positives obtained as a result of the turkeys being previously vaccinated with egg-grown virus vaccines (Newcastle disease virus). The details of the technique and the importance of antigen preparation are discussed.

            Descriptors:  antigens, viral isolation and purification, enzyme linked immunosorbent assay, immunoenzyme techniques, influenza A virus avian isolation and purification, antibodies, anti idiotypic isolation and purification, chick embryo, chromatography, affinity, chromatography, gel, fowl plague diagnosis, horseradish peroxidase, avian immunology, rabbits immunology, turkeys immunology.

Adair, B.M., K. Burns, M.S. McNulty, and D. Todd (1990). A study of ELISA systems incorporating pooled viral and Mycoplasma antigen preparations for antibody screening of chicken sera. Avian Pathology 19(2): 263-278.  ISSN: 0307-9457.

            NAL Call Number:  SF995.A1A9

            Descriptors:  avian influenza virus, screening techniques, ELISA, Mycoplasma synoviae, Mycoplasma gallisepticum, Gallus gallus, chickens.

Al Attar, M., K. Nielsen, and W.R. Mitchell (1981). The application of the soluble antigen fluorescent antibody test for the diagnosis of avian influenza. Canadian Journal of Comparative Medicine Revue Canadienne De Medecine Comparee 45(2):  140-6.  ISSN: 0008-4050.

            NAL Call Number:  41.8 C162

            Abstract:  The application of the soluble antigen fluorescent test as a tool for serological investigation of influenza type A infection in wild birds was studied. The soluble antigen fluorescent antibody test is basically an indirect fluorescent antibody test except that an artificial matrix of cellulose acetate discs is used as a substrate for antigen and the test results are scanned and recorded by a fluorometer. THe influenza type A soluble antigen fluorescent antibody was obtained from concentrated and detergent disrupted virus particles, absorbed onto cellulose acetate discs. Anti-influenza sera were prepared in pheasants and ducks to A/turkey/Ontario/6118/67 and in pigeons to A/turkey/Ontario/6213/68. The antigen-antibody complex was detected by specific staining with monovalent or polyvalent fluorescein isothiocyanate conjugated rabbit anti-avian immunoglobulins. The soluble antigen fluorescent antibody test is a sensitive technique for the detection of specific influenza A antibodies in several avian species, and could be adapted for use in large scale surveys.

            Descriptors:  antibodies, viral analysis, fluorescent antibody technique, fowl plague diagnosis, influenza A virus avian immunology, antigens, viral, birds, hemagglutination inhibition tests veterinary, poultry, solubility.

Al Natour, M.Q. and M.N. Abo Shehada (2005). Sero-prevalence of avian influenza among broiler-breeder flocks in Jordan. Preventive Veterinary Medicine 70(1-2): 45-50.

            NAL Call Number:  SF601.P7

            Abstract:  Thirty blood samples were collected randomly from each of the 38 breeder-broiler farms in Jordan. Serum samples were examined using indirect ELISA for specific antibodies to avian influenza virus. The overall true flock-level sero-prevalence of avian influenza was 71% (95% CI: 55,83). Positive flocks had 2-30 sero-positive chickens and half of flocks had >20 sero-positive birds. The number of sero-positive flocks varied in the studied localities with more sero-positives in farms located within the migratory route of migratory wild fowl. The examined broiler-breeder flocks had no clinical signs, or noticeable decrease in egg production; mortalities were within the normal range (0.1-1%). The number of positive sera/flock correlated with flock size. There were a no significant (Pearsons r = 0.21, p = 0.21) correlation between positive flocks and age. A non-pathogenic AI virus infects broiler-breeder farms in Jordan. Wild local and migrating birds might promote the further spread of this virus in Jordan and other countries.

            Descriptors:  avian influenza, poultry, viral diseases, broiler-breeder, ELISA, age influence, Jordan.

Alexander, D.J. (2000). Highly pathogenic avian influenza. In: Manual of Standards for Diagnostic Tests and Vaccines. List A and B Diseases of Mammals, Birds and Bees, 4th edition, p. 212-220. ISBN: 92-9044-510-6.

            NAL Call Number:  SF771.M36 2000

            Descriptors:  fowl plague virus, influenza virus A, immunization, diagnosis, techniques, mortality, pathogenicity, diagnostic tests, manual of standards, vaccines, Gallus gallus, poultry.

Allan, G.M. and M.S. McNulty (1985). A direct immunofluorescence test for the rapid detection of avian influenzavirus antigen in tissue impression smears. Avian Pathology 14(4): 449-460.  ISSN: 0307-9457.

            NAL Call Number:  SF995.A1A9

            Descriptors:  immunofluorescent test, avian influenza virus, diagnosis, techniques, detection, poultry.

Allan, W.H. (1981). Diagnostic procedures--response. In: Proceedings of the First International Symposium on Avian Influenza, Beltsville, Maryland, USA, 167-171 p.

            NAL Call Number: aSF995.6.I6I5 1981a

            Descriptors: avian influenza virus, diagnostic procedures.

Allan, W.H., D.J. Alexander, B.S. Pomeroy, and G. Parsons (1977). Use of virulence index tests for avian influenza viruses. Avian Diseases 21(3): 359-63.  ISSN: 0005-2086.

            NAL Call Number:  41.8 Av5

            Abstract:  The intravenous and intracerebral pathogenicity index tests normally used for Newcastle disease virus isolates were used to measure the virulence of 13 avian influenza viruses. The tests allowed quantitative measurements of the virulence of the avian influenza viruses, and the results confirmed the range in virulence, between the two extremes, of the avian influenza viruses and demonstrated the lack of correlation between virulence and antigenic type.

            Descriptors:  chickens, fowl plague etiology, influenza A virus avian pathogenicity, antigens, viral analysis, brain, chick embryo, fowl plague mortality, avian immunology, injections, intravenous, methods, virulence.

Apisarnthanarak, A., R. Kitphati, K. Thongphubeth, P. Patoomanunt, P. Anthanont, W. Auwanit, P. Thawatsupha, M. Chittaganpitch, S. Saeng Aroon, S. Waicharoen, P. Apisarnthanarak, G.A. Storch, L.M. Mundy, and V.J. Fraser (2004). Atypical avian influenza (H5N1). Emerging Infectious Diseases 10(7): 1321-4.  ISSN: 1080-6040.

            NAL Call Number:  RA648.5.E46

            Abstract:  We report the first case of avian influenza in a patient with fever and diarrhea but no respiratory symptoms. Avian influenza should be included in the differential diagnosis for patients with predominantly gastrointestinal symptoms, particularly if they have a history of exposure to poultry.

            Descriptors:  gastrointestinal diseases physiopathology, influenza physiopathology, influenza A virus, avian pathogenicity, adult, chickens virology, fatal outcome, gastrointestinal diseases virology, health personnel, influenza virology, influenza, avian transmission, influenza, avian virology, poultry diseases transmission, poultry diseases virology.

Astorga, R.J., L. Leon, M.J. Cubero, A. Arenas, A. Maldonado, M.C. Tarradas, and A. Perea (1994). Avian influenza in wild waterfowl and shorebirds in the Donana National Park: Serological survey using the enzyme-linked immunosorbent assay. Avian Pathology 23(2): 339-344.  ISSN: 0307-9457.

            NAL Call Number:  SF995.A1A9

            Abstract:  The indirect ELISA was used to detect antibodies to influenzavirus A in the sera of wildfowl from the Donana National Park. Of the 712 birds examined, 44 (6.2%) were seropositive. Positive birds belonged to 10 of the 13 species studied. Infection rates varied widely: spoonbill (Platalea leucorodia, 32.2%), mallard (Anas platyrhynchos, 9.9%), gadwall (Anas strepera, 8.6%), red-crested pochard (Netta rufina, 8.1%), pochard (Aythya ferina, 6.4%), shoveler (Anas clypeata, 5%), great crested grebe (Podiceps cristatus, 4.3%), avocet (Recurostra avosetta, 3.1%), grey heron (Ardea cinerea, 3.1%) and coot (Fulica atra, 0.8%). Although infection rates were not high, the wide range of avian species susceptible to influenzavirus A suggests circulation of the virus amongst wildfowl at Donana.

            Descriptors:  enzymology, immune system, infection, methods and techniques, pathology, veterinary medicine, diagnostic method ELISA epidemiology.

Australia Commonwealth Scientific and Industrial Research Organisation (1988). Australian Standard Diagnostic Techniques for Animal Diseases. Nos. 1-51, ISBN: 0643040765.

            Descriptors:  standard diagnostic techniques, animal diseases, Australia, booklet series.

Barli Maganja, D., U. Krapez, S. Manko, I. Toplak, J. Grom, P. Hostnik, and O.Z. Rojs (2004). New approaches in diagnosis and typing of viruses causing diseases in poultry. Praxis Veterinaria (Zagreb) 52(1/2): 19-26.  ISSN: 0350-4441.

            Abstract:  In the last two decades, various molecular biological methods were introduced in diagnostic virology. They are used for the rapid detection of viral nucleic acids, genetic characterization of the pathogens responsible for many viral infections and tracking of the origin and spread of viruses. In this review, the application of molecular biology methods, particularly the combined approach of amplifying defined fragments of viral genomes, using the polymerase chain reaction and subsequent nucleotide sequencing analysis, is described. Emphasis is placed on some of the few important viruses causing economically important diseases in poultry, like Newcastle disease virus, avian influenza virus, infectious bursal disease virus and chicken anaemia virus.

            Descriptors:  avian infectious bursitis, diagnosis, diagnostic techniques, DNA sequencing, fowl diseases, genomes, influenza, methodology, molecular biology, Newcastle disease, polymerase chain reaction, poultry, reviews, avian influenza virus, chicken anaemia virus, fowls, infectious bursal disease virus, Newcastle disease virus.

Barr, D.A. and M.D. O'Rourke (1993). Avian influenza: pathology, virology and serology. In: L.A. Corner and T.J. Bagust (editor), Australian Standard Diagnostic Techniques for Animal Diseases,  East Melbourne, Vic. 3002 Australia, 6 p. ISBN: 0-643-05243-7.

            NAL Call Number:  SF772.6.A97 1993

            Descriptors:  standard diagnostic techniques, avian influenza virus, Gallus gallus, Australia.

Barr, D.A. and M.D. O' Rourke (1987). Virulent avian influenza. In: Australian Standard Diagnostic Techniques for Animal Diseases, Vol. 51, 14 p.

            NAL Call Number:  SF771.A8A97 no.51

            Descriptors:  avian influenza virus, standard diagnostic techniques, Australia.

Beard, C.W. (1970). Avian influenza antibody detection by immunodiffusion. Avian Diseases 14(2): 337-41.  ISSN: 0005-2086.

            NAL Call Number:  41.8 Av5

            Descriptors:  antibodies analysis, immunodiffusion, influenza diagnosis, poultry diseases diagnosis, antigens isolation and purification, chickens, hemagglutination inhibition tests, influenza immunology, orthomyxoviridae immunology, orthomyxoviridae isolation and purification, poultry diseases immunology,  turkeys.

Beard, C.W. and B.C. Easterday (1975). Isolierung und Identifizierung eines Vogel-Influenza-Virus mit dem Hamagglutinin des Geflugelpestvirus. [Isolation and identification of an avirulent avian influenza virus with the hemagglutinin of fowl plague virus].  Proceedings of the 5th World Veterinary Poultry Association Congress, Munich 1973 1: 725-736.

            Descriptors:  avian influenza virus, diagnosis, fowl plague virus, identification, isolation.

Beard, C.W. and B.C. Easterday (1973). A-Turkey-Oregon-71, an avirulent influenza isolate with the hemagglutinin of fowl plague virus. Avian Diseases 17(1): 173-81.  ISSN: 0005-2086.

            NAL Call Number:  41.8 Av5

            Descriptors:  hemagglutinins viral, influenza A virus avian immunology, orthomyxoviridae immunology, antibodies, viral analysis, chickens, fowl plague immunology, fowl plague microbiology, hemagglutination inhibition tests, immunization, influenza immunology, influenza microbiology, influenza veterinary, avian enzymology, avian isolation and purification, neuraminidase analysis, neutralization tests, orthomyxoviridae enzymology, orthomyxoviridae isolation and purification, poultry diseases immunology, poultry diseases microbiology, virulence.

Becht, H. (1968). Properties of erythrocytes stabilized with sulfosalicylic acid and their use in an indirect hemagglution test with influenza virus RNP-antigen. Journal of Immunology 101(1): 18-22.  ISSN: 0022-1767.

            NAL Call Number:  448.8 J8232

            Descriptors:  antigens, erythrocytes, hemagglutination tests, indicators and reagents, orthomyxoviridae, salicylic acids, sulfonic acids, complement fixation tests, immune sera, influenza A virus avian, nucleoproteins, sheep.

Becht, H. and B. Malole (1975). Comparative evaluation of different fixation procedures and different coupling reagents for the demonstration of influenza virus-specific antibodies by the indirect hemagglutination test. Medical Microbiology and Immunology 162(1): 43-53.  ISSN: 0300-8584.

            Abstract:  The indirect hemagglutination technique has been improved by fixing the carrier erythrocytes successively with glutaraldehyde and sulfosalicylic acid. Sensitization by covalent conjugation of influenza virus antigens to the erythrocytes with various coupling reagents, which resulted in stable and highly sensitive test cells, has been defined. An economical affinity chromatography procedure using antibody-coated agarose has been developed to prepare sufficiently pure antigens from fowl plague virus-infected choriollantoic membranes.

            Descriptors:  antibodies, viral analysis, erythrocytes immunology, hemagglutination tests methods, antibody specificity, blood preservation, chromatography, affinity, cytological techniques, glutaral, hemagglutinins viral isolation and purification, influenza A virus avian immunology, salicylic acids.

Beck, J.R., D.E. Swayne, S. Davison, S. Casavant, and C. Gutierrez (2003). Validation of egg yolk antibody testing as a method to determine influenza status in white leghorn hens. Avian Diseases 47(Special Issue): 1196-1199.  ISSN: 0005-2086.

            NAL Call Number:  41.8 Av5

            Abstract:  Determination of the avian influenza (AI) status of a flock has traditionally been done by detection of serum antibodies. However, for many diseases, detection of antibodies in egg yolk has been effective in monitoring the disease status of laying flocks. This study compared the utility of egg yolk vs. serum for determining AI status in laying hen flocks. Specific-pathogen-free white leghorn hens were inoculated via the respiratory tract with a low-pathogenic H7N2 AI virus or sterile allantoic fluid or subcutaneously with an inactivated oil emulsion vaccine produced from the same AI virus or normal allantoic fluid. Antibody levels were determined by the agar gel immunodiffusion (AGID) test, the hemagglutination-inhibition (HI) test, and the enzyme-linked immunosorbent assay (ELISA). Anti-influenza antibodies were detected in sera of all live virus-inoculated hens by day 7 postinoculation (PI) (AGID and ELISA tests), but detection of antibodies in egg yolk was delayed by a few days, with all being positive by day 14 PI. Sera from all vaccinated hens were positive by day 14 PI (AGID and ELISA tests), and egg yolk was positive by day 18 PI. The HI test was less sensitive than the ELISA and AGID tests in detecting anti-influenza antibodies in both sera and yolk. Serum and yolk from all control birds remained negative throughout the study. These studies show that currently used serologic tests can detect antibodies in serum and yolk samples from hens exposed to live AI virus or from those that have been vaccinated. Antibody is detected earlier in the serum than in the yolk and antibody is detected earlier from birds exposed to a live infection compared to birds vaccinated with an inactivated oil emulsion vaccine.

            Descriptors:  animal husbandry, immune system, infection, avian influenza, infectious disease, viral disease, ELISA immunologic techniques, laboratory techniques, agar gel immunodiffusion test, agid test, egg yolk antibody testing, hemagglutination inhibition test, influenza status, laying flock disease status.

Berinstein, A., B.S. Seal, and D.L. Suarez (2002). Heteroduplex mobility assay for detection of new avian influenza virus variants. Avian Diseases 46(2): 393-400.  ISSN: 0005-2086.

            NAL Call Number:  41.8 Av5

            Abstract:  Highly pathogenic avian influenza (HPAI) in poultry causes high morbidity and mortality, and it is a List A disease of the Office International des Epizooties. An outbreak of HPAI in commercial poultry not only causes direct disease losses but often results in trade restrictions for the affected country. Because HPAI viruses can mutate from H5 and H7 low pathogenic avian influenza viruses, it is necessary to monitor and control even the low pathogenic form of the virus. We report a practical approach for screening large numbers of isolates that uses amplification by reverse transcriptase-polymerase chain reaction of a segment of the hemagglutinin (HA) gene (536-560 bp) of H7 avian influenza viruses followed by the heteroduplex mobility assay (HMA). The HMA test compares the amplified polymerase chain reaction product from unknown samples with reference isolates, which allows the identification of new variants. The HMA test results were compared with sequence analysis of the isolates used in the study. On the basis of the HMA, we could identify several new variant viruses present in the live bird markets in the northeastern United States. New strains gave a distinct pattern of bands in the gels in accordance with the different heteroduplexes formed when their HA region amplification products were incubated together with the same amplification product of a reference strain. These differences correlate with phylogenetic analysis from sequence data.

            Descriptors:  animal husbandry, infection, molecular genetics, avian influenza virus infection, infectious disease, viral disease, gene sequencing cycle DNA sequencing, sequencing method, heteroduplex mobility assay bioassay method, phylogenetic analysis genetic method, reverse transcriptase polymerase chain reaction genetic method, polymerase chain reaction, morbidity mortality.

Brugh, M. and C.W. Beard (1980). Collection and processing of blood samples dried on paper for microassay of Newcastle disease virus and avian influenza virus antibodies. American Journal of Veterinary Research 41(9): 1495-8.  ISSN: 0002-9645.

            NAL Call Number:  41.8 Am3A

            Abstract:  A practical method for collection and processing of dried whole blood samples on filter paper was developed to facilitate large-scale testing programs for Newcastle disease virus and avian influenza virus antibodies. A modified paper punch was used to cut and place dried blood samples simultaneously in a standard 96-well microlate for elution of antibody. Twelve eluted samples were simultaneously transferred to another microplate for the hemagglutination-inhibition (HI) microtest. Similar HI titers were obtained with simultaneously collected serum and dried blood samples. Minor differences were not considered of practical importance in diagnostic serologic studies. Dried blood titers were not markedly affected by method of drying (37 C for 2 hours or 26 C for 4 hours), by storage for 24 hours before drying, or by storage of dried samples at 4 C for 28 days or 30 C for 14 days. Blood dried on paper was a satisfactory sample for assay of HI antibodies to Newcastle disease virus and avian influenza virus.

            Descriptors:  antibodies, viral analysis, blood specimen collection veterinary, chickens immunology, influenza A virus avian immunology, Newcastle disease virus immunology, blood specimen collection instrumentation, hemagglutination inhibition tests, paper.

Capua, I., G. Cattoli, and S. Marangon (2004). DIVA--a vaccination strategy enabling the detection of field exposure to avian influenza. Developmental Biology (Basel) 119: 229-33.  ISSN: 1424-6074.

            Abstract:  The present paper reports on the development, validation and field application of a control strategy for avian influenza infections in poultry. The "DIVA" (Differentiating Infected from Vaccinated Animals) strategy is based on the use of an inactivated oil emulsion vaccine containing the same haemagglutinin (H) subtype as the challenge virus, but a different neuraminidase (N). The possibility of using the heterologous N subtype, to differentiate between vaccinated and naturally infected birds, was investigated through the development of an "ad hoc" serological test based on the detection of specific anti-N antibodies. This test is based on an indirect fluorescent antibody assay, using as an antigen a baculovirus expressing recombinant N proteins. The vaccination strategy has been tested in the laboratory and shown to be efficacious both against challenge with highly pathogenic AI viruses and with low pathogenicity AI viruses, ensuring clinical protection, reduction of duration and titre of shedding. In addition, vaccination resulted in an increased resistance to infection. The companion diagnostic tests directed to the detection of anti-N1 and anti-N3 antibodies have been validated in the laboratory and using field samples. The serological assay showed an "almost perfect agreement" (Kappa value) with the HI test, with relative sensitivity and specificity values of 98.1 and 95.7, respectively. The results of the present investigation suggest that the "DIVA" control strategy may represent a tool to support the eradication of avian influenza infections in poultry.

            Descriptors:  animals, viral blood antibodies, viral immunology antibodies, genetic engineering, avian influenza A virus enzymology, avian influenza diagnosis, avian influenza prevention and control, neuraminidase genetics, poultry, sensitivity and specificity, veterinary serologic tests, marker vaccines, viral vaccines immunology, virus shedding.

Cattoli, G., A. Drago, S. Maniero, A. Toffan, E. Bertoli, S. Fassina, C. Terregino, C. Robbi, G. Vicenzoni, and I. Capua (2004). Comparison of three rapid detection systems for type A influenza virus on tracheal swabs of experimentally and naturally infected birds. Avian Pathology 33(4): 432-7.  ISSN: 0307-9457.

            NAL Call Number:  SF995.A1A9

            Descriptors:  influenza A virus, avian isolation and purification, avian diagnosis, poultry diseases diagnosis, poultry diseases virology, immunoenzyme techniques methods, avian classification, avian genetics, reverse transcriptase polymerase chain reaction methods, sensitivity and specificity, trachea virology, turkeys.

Cattoli, G., C. Terregino, V. Brasola, J.F. Rodriguez, and I. Capua (2003). Development and preliminary validation of an ad hoc N1-N3 discriminatory test for the control of avian influenza in Italy. Avian Diseases 47(Special Issue): 1060-1062.  ISSN: 0005-2086.

            NAL Call Number:  41.8 Av5

            Abstract:  The development of a discriminatory test, based on the differentiation between N1 and N3 antibodies, to be used in the framework of a vaccination program, based on vaccination with a heterologous H7N3 inactivated vaccine against the Italian H7N1 field virus, is reported. The indirect immunofluorescence antibody (iIFA) assay was based on the expression of the N1 protein in a baculovirus system. HighFive(R) insect cells were transfected with the recombinant virus and used as an antigen in the iIFA test. Preliminary validation on 608 turkey sera yielded relative sensitivity and specificity of 98.1% and 95.7%, respectively, when compared to the HI test with an almost perfect agreement between the two methods (Kappa value = 0.93). It is concluded that the iIFA test is a valid tool for monitoring avian influenza infection in a vaccinated population.

            Descriptors:  animal husbandry, immune system, infection, antibody differentiation test, immunologic techniques, immunofluorescence antibody assay, bioassay techniques, laboratory techniques, vaccination, clinical techniques, influenza control.

Ceron, H.M., V.H. Rodriguez, M.A. Hernandez, R.K. Blasco, J. Garcia Garcia, and R.G. Webster ( 1996). Estudios basicos de las cepas de desafio de influenza aviar y evaluacion de la prueba de patogenicidad. [Basic studies with avian influenza challenge viruses, and evaluation of the pathogenicity test]. Proceedings of the Western Poultry Diseases Conference 45: 50-51.

            NAL Call Number:  SF995.W4

            Descriptors:  avian influenza virus, basic studies, challenge, pathogenicity test.

Chu, H.P., N.M. Barhouma, S. Eid, J.R. Fuller, and M.K. Fuller (1975). Egg yolk haemagglutination-inhibition tests for Newcastle disease and avian influenza. Proceedings of the 5th World Veterinary Poultry Association Congress, Munich 1973 II: 1014-1019.

            Descriptors:  diagnosis, hemagglutination inhibition test, Newcastle disease, avian influenza, egg yolk, poultry.

Collins, R.A., L.S. Ko, K.L. So, T. Ellis, L.T. Lau, and A.C.H. Yu (2003). A NASBA method to detect high- and low-pathogenicity H5 avian influenza viruses. Avian Diseases 47(Special Issue): 1069-1074.  ISSN: 0005-2086.

            NAL Call Number:  41.8 Av5

            Abstract:  Nucleic acid sequence-based amplification (NASBA) allows the rapid amplification of specific regions of nucleic acid obtained from a diverse range of sources. It is especially suitable for amplifying RNA sequences. A NASBA technique was developed that allows the detection of avian influenza A subtype H5 from allantoic fluid harvested from inoculated chick embryos. The amplified viral RNA is detected by electrochemiluminescence. The described NASBA technique is a specific, rapid, and sensitive method of detection of influenza A subtype H5 viruses. More importantly, it can be used to distinguish high- and low-pathogenicity strains of the H5 subtype.

            Descriptors:  immune system, infection, molecular genetics, electrochemiluminescence, immunologic techniques, laboratory techniques, nucleic acid amplification, genetic techniques, nucleic acid sequence based amplification, NASBA.

Collins, R.A., L.S. Ko, K.Y. Fung, K.Y. Chan, J. Xing, L.T. Lau, and A.C.H. Yu (2003). Rapid and sensitive detection of avian influenza virus subtype H7 using NASBA. Biochemical and Biophysical Research Communications 300(2): 507-515.  ISSN: 0006-291X.

            NAL Call Number:  442.8 B5236

            Abstract:  Nucleic acid sequence-based amplification with electrochemiluminescent detection (NASBA/ECL) is an isothermal technique allowing rapid amplification and detection of specific regions of nucleic acid from a diverse range of sources. It is especially suitable for amplifying RNA. A NASBA/ECL technique has been developed allowing the detection of RNA from avian influenza virus subtype H7 derived from allantoic fluid harvested from inoculated chick embryos and from cell cultures. Degenerate amplification primers and amplicon capture probes were designed enabling the detection of low and highly pathogenic avian influenza of the H7 subtype from the Eurasian and North American lineages and the Australian sub-lineage. The NASBA/ECL technique is specific for subtype H7 and does not cross-react with other influenza subtypes or with viruses containing haemagglutinin-like genes. The assay is 10- to 100-fold more sensitive than a commercially available antigen capture immunoassay system. The NASBA/ECL assay could be used in high throughput poultry screening programmes.

            Descriptors:  molecular genetics, influenza, diagnosis, respiratory system disease, viral disease, nucleic acid based amplification with electrochemiluminescent detection genetic techniques, laboratory techniques.

Collins, R.A., L.S. Ko, K.L. So, T. Ellis, L.T. Lau, and A.C.H. Yu (2002). Detection of highly pathogenic and low pathogenic avian influenza subtype H5 (Eurasian lineage) using NASBA. Journal of Virological Methods 103(2): 213-225.  ISSN: 0166-0934.

            NAL Call Number:  QR355.J6

            Abstract:  Nucleic acid sequence-based amplification (NASBA) is a technique that allows the rapid amplification of specific regions of nucleic acid obtained from a diverse range of sources. It is especially suitable for amplifying RNA sequences. A NASBA technique has been developed that allows the detection of avian influenza A subtype H5 from allantoic fluid harvested from inoculated chick embryos. The amplified viral RNA is detected by electrochemiluminescence. The NASBA technique described below is rapid and specific for the identification of influenza A subtype H5 viruses of the Eurasian lineage. More importantly, it can be used to distinguish highly pathogenic and low pathogenic strains of the H5 subtype.

            Descriptors:  human medicine, infection, methods and techniques, molecular genetics, DNA sequencing analytical method, recombinant DNA technology, sequencing techniques, electrochemiluminescence technique analytical method, applications, description, molecular method, nucleic acid sequence based amplification technique molecular biology techniques and chemical characterization, applications, description, molecular method, reverse transcriptase polymerase chain reaction molecular method, polymerase chain reaction, diagnostics, pathogenicity, viral genetics, virological methodologies applications, virulence.

Davison, S., A.F. Ziegler, and R.J. Eckroade (1998). Comparison of an antigen-capture enzyme immunoassay with virus isolation for avian influenza from field samples. Avian Diseases 42(4): 791-795.  ISSN: 0005-2086.

            NAL Call Number:  41.8 Av5

            Abstract:  The standard tests used to detect avian influenza (AI) viral infection include virus isolation from tissues of the infected birds and the detection of Al antibody in blood or egg yolk. A new application of an existing human test to rapidly detect the presence of any influenza A virus is now possible. A commercially available antigen-capture enzyme immunoassay (AC-EIA), developed for the detection of influenza A in humans, was tested for relative sensitivity and specificity and for speed of use  in diagnosing nonpathogenic H7N2 AI in naturally infected poultry. During the recent nonpathogenic H7N2 AI epornitic, the AC-EIA was used for rapid diagnosis and quarantine decisions. Between February and August 1997, 1524 samples from 295 commercial layer, pullet, and broiler flocks were submitted to the Laboratory of Avian Medicine and Pathology, New Bolton Center, for AI virus isolation and testing by AC-EIA. The relative specificity of the AC-EIA was 100% and the relative sensitivity was  79%. We believe that the AC-EIA will be a useful adjunct to standard AI diagnostic tests.

            Descriptors:  infection, methods and techniques, veterinary medicine, avian influenza, detection, respiratory system disease, viral disease, antigen capture enzyme immunoassay comparison, diagnostic method, virus isolation comparison, diagnostic method.

de Boer, G.F., W. Back, and A.D. Osterhaus (1990). An ELISA for detection of antibodies against influenza A nucleoprotein in humans and various animal species. Archives of Virology 115(1-2): 47-61.  ISSN: 0304-8608.

            NAL Call Number:  448.3 Ar23

            Abstract:  A double antibody sandwich blocking ELISA, using a monoclonal antibody (MAb) against influenza A nucleoprotein (NP) was developed to detect antibodies against influenza. Collections of serum samples were obtained from human and various animal species. All influenza A subtypes induced antibodies against hemagglutinins and NP. A close correlation between titers of the hemagglutination inhibition (HI) test and the NP-ELISA was seen. Antibodies against influenza NP were demonstrated in serum samples from humans, ferrets, swine, horses, chickens, ducks, guinea pigs, mice, and seals. The serum samples were collected at intervals during prospective epidemiological studies, from experimental and natural infections, and vaccination studies. The decline of maternal antibodies was studied in swine and horses. The NP-ELISA enables rapid serological diagnosis and is suited for influenza A antibody screening, especially in species which harbor several influenza subtypes. The HI and neuraminidase inhibition tests, however, must still be used for subtyping.

            Descriptors:  antibodies, viral analysis, enzyme linked immunosorbent assay, influenza A virus immunology, nucleoproteins immunology, orthomyxoviridae infections immunology, viral core proteins immunology, ferrets, hemagglutination inhibition tests, horses, avian immunology, human immunology, porcine immunology, orthomyxoviridae infections veterinary, poultry, prospective studies, Rodentia, seals, species specificity, specific pathogen free organisms, swine, vaccination.

Durham, S. (2003). A new, rapid test for avian influenza. Agricultural Research 51(2): 9.  ISSN: 0002-161X.

            Online:  www.ars.usda.gov/is/AR/

            NAL Call Number:  1.98 Ag84

            Descriptors:  avian influenza virus, influenza, poultry diseases, laboratory tests, rapid methods, reverse transcription, polymerase chain reaction, pathogenicity, United States.

Dybkaer, K., M. Munch, K.J. Handberg, and P.H. Jorgensen (2004). Application and evaluation of RT-PCR-ELISA for the nucleoprotein and RT-PCR for detection of low-pathogenic H5 and H7 subtypes of avian influenza virus. Journal of Veterinary Diagnostic Investigation, Official Publication of the American Association of Veterinary Laboratory Diagnosticians, Inc 16(1): 51-6.  ISSN: 1040-6387.

            NAL Call Number:  SF774.J68

            Abstract:  Three 1-tube Reverse Transcriptase Polymerase Chain Reactions (RT-PCR) directed against the genes encoding the nucleoprotein (NP) and the H5 and H7 hemagglutinin (HA) gene, respectively, were used for detection of avian influenza virus (AIV) in various specimens. A total of 1,040 samples originating from chickens experimentally infected with 2 different low pathogenic avian influenza viruses, from domestic ducks and from wild aquatic birds were examined. The outcome of 1) the universal AIV RT-PCR including a PCR-enzyme-linked immunosorbent assay (ELISA) procedure directed against NP (NP RT-PCR-ELISA) and 2) the subtype specific RT-PCR for H5 and H7 were compared to the results obtained by inoculation of the same specimens into the allantoic cavity of embryonated specific pathogen free (SPF) hen's eggs. Using inoculation in SPF fowl eggs as standard the sensitivity of the NP RT-PCR-ELISA and the RT-PCR for H5 or H7 was 91% and 94%, and the corresponding specificity 98% and 96%. In comparison with inoculation into eggs an additional of 9 samples were positive by NP RT-PCR-ELISA and 13 samples were positive by RT-PCR for one of the HA subtypes. Hence, the 3 RT-PCR procedures described are fast, sensitive and specific for detecting AIV and subtyping H5 and H7 and they are obvious alternatives when testing large numbers of samples.

            Descriptors:  enzyme linked immunosorbent assay veterinary, hemagglutinins genetics, influenza veterinary, influenza A virus, avian classification, avian genetics, avian pathogenicity, nucleoproteins chemistry, avian isolation and purification, nucleoproteins genetics, poultry diseases virology, reverse transcriptase polymerase chain reaction veterinary, antibodies, viral blood, chick embryo, chickens, ducks, enzyme linked immunosorbent assay methods, hemagglutination inhibition tests veterinary, influenza diagnosis, influenza virology, RNA, viral chemistry, viral genetics, reverse transcriptase polymerase chain reaction methods, sequence analysis, DNA, virulence.

Dybkaer, K., M. Munch, K.J. Handberg, and P.H. Jorgensen (2003). RT-PCR-ELISA as a tool for diagnosis of low-pathogenicity avian influenza. Avian Diseases 47(Special Issue): 1075-1078.  ISSN: 0005-2086.

            NAL Call Number:  41.8 Av5

            Abstract:  A one-tube reverse transcriptase/polymerase chain reaction coupled with an enzyme-linked immunosorbent assay (RT-PCR-ELISA) was developed for the rapid detection of avian influenza virus (AIV) in clinical specimens. A total of 419 swab pools were analyzed from chickens experimentally infected with low-pathogenicity AIV, from wild aquatic birds, and from domestic ducks. The AIV was detected in 32 swab pools by RT-PCR-ELISA compared to 23 by virus isolation (VI) in embryonated specific pathogen free (SPF) chicken eggs. Thus, 39% more specimens were positive by RT-PCR-ELISA than by VI. Two of the twenty-three VI-positive specimens were negative when tested by RT-PCR-ELISA. The diagnostic sensitivity and specificity of the RT-PCR-ELISA was 91% and 97%, respectively, using VI in SPF eggs as the gold reference standard.

            Descriptors:  infection, molecular genetics, reverse transcriptase polymerase chain reaction ELISA clinical techniques, diagnostic techniques, genetic techniques, immunologic techniques, laboratory techniques, clinical specimens.

Elbers, A.R., B. Kamps, and G. Koch (2005). Diagnostische mogelijkheden van pathologische laesies bij pluimvee voor het opsporen van uitbraken tijdens de klassieke vogelpestepidemie in Nederland in 2003. [Diagnostic approaches to pathological lesions in fowl to determine outbreaks during the classical avian influenza epidemic in the Netherlands in 2003]. Tijdschrift Voor Diergeneeskunde 130(1): 14.  ISSN: 0040-7453.

            NAL Call Number:  41.8 T431

            Descriptors:  disease outbreaks veterinary, avian influenza epidemiology, avian influenza diagnosis, avian influenza pathology, Netherlands epidemiology, poultry.

Elbers, A.R., B. Kamps, and G. Koch (2004). Performance of gross lesions at postmortem for the detection of outbreaks during the avian influenza A virus (H7N7) epidemic in The Netherlands in 2003. Avian Pathology 33(4): 418-22.  ISSN: 0307-9457.

            NAL Call Number:  SF995.A1A9

            Descriptors:  disease outbreaks veterinary, influenza A virus, avian, influenza, avian diagnosis, poultry diseases epidemiology, chickens, edema pathology, edema veterinary, epidemiology, pathology, neck pathology, Netherlands epidemiology, peritonitis pathology, peritonitis veterinary, poultry diseases diagnosis, poultry diseases pathology, tracheitis pathology, tracheitis veterinary.

Elkin, V.S., R.I.A. Podcherniaeva, E.V. Sidorenko, F.E. Sadykhova, and T.A. Ignatenko (1988). Vyiavlenie antitel s pomoshch'iu rekombinantnykh shtammov virusov grippa v syvorotakh razlichnykh vidov ptits, tsirkuliruiushchikh na territorii Ukrainskoi i Azerbaidzhanskoi SSR. [Detection of antibodies using recombinant strains of influenza viruses in the sera of various species of birds circulating in the territories of the Ukrainian and Azerbaijan SSRs]. Voprosy Virusologii 33(6): 674-6.  ISSN: 0507-4088.

            NAL Call Number:  448.8 P942

            Abstract:  Examinations of blood sera from different species of birds trapped in the Ukrainian and Azerbaijan SSRs using diagnostic preparations from the influenza A/sea gull/Maryland/704/77 virus strain and a recombinant R117 derived from it revealed the presence of antibodies to hemagglutinin H13. The diagnostic preparation produced from the recombinant strain was found to be more active in the detection of antibodies in avian sera.

            Descriptors:  antibodies, viral analysis, birds immunology, influenza A virus avian immunology, recombination, genetic, Azerbaijan, hemagglutination inhibition tests veterinary, avian genetics, Ukraine.

Ellis, T.M., L. Hustas, J.S. MacKenzie, and I.M. Watson (1988). Rapid detection of group specific influenza A virus antigen [avian influenza; fowl plague]. Australian Veterinary Journal 65(11): 357-358.  ISSN: 0005-0423.

            NAL Call Number:  41.8 Au72

            Descriptors:  chickens, avian influenza virus, antigens, diagnosis, birds, domestic animals, domesticated birds, Galliformes, immunological factors, immunology, influenza virus, livestock, poultry, useful animals, viruses.

Fatunmbi, O.O., J.A. Newman, V. Sivanandan, and D.A. Halvorson (1989). A broad-spectrum avian influenza subtype antigen for indirect enzyme-linked immunosorbent assay. Avian Diseases 33(2): 264-9.  ISSN: 0005-2086.

            NAL Call Number:  41.8 Av5

            Abstract:  A broad-spectrum viral antigen for the detection of avian-influenza-virus-specific antibodies, using the indirect enzyme-linked immunosorbent assay (ELISA), was identified. Purified and disrupted antigens were used, which helped to increase the sensitivity of the assay. All of the antigens tested were able to detect antibodies to homologous and heterologous viruses to varying degrees. The H9N2 antigen was the best single antigen to use in the ELISA to screen for avian influenza virus antibodies. It detected antibodies against six viruses as early as day 4 postinfection.

            Descriptors:  antibodies, viral analysis, antigens, viral immunology, enzyme linked immunosorbent assay, fowl plague immunology, influenza A virus avian immunology, antigens, viral analysis, turkeys immunology.

Fatunmbi, O.O., J.A. Newman, V. Sivanandan, and D.A. Halvorson (1992). Efficacy of avridine and liposomes as adjuvants for avian influenza virus antigens in turkeys.  Avian Pathology 21(2): 225-237.  ISSN: 0307-9457.

            NAL Call Number:  SF995.A1A9

            Descriptors:  disease control, adjuvants, avian influenza virus antigens, vaccines, turkeys.

Forsyth, W.M., D.C. Grix, and C.A. Gibson (1993). Diagnosis of highly pathogenic avian influenza in chickens: Bendigo 1992.  Australian Veterinary Journal 70(3): 118-9.  ISSN: 0005-0423.

            NAL Call Number:  41.8 Au72

            Descriptors:  disease outbreaks veterinary, fowl plague epidemiology, influenza A virus avian immunology, antibodies, viral analysis, chickens, Victoria epidemiology.

Hadjiev, G., V. Bumbarov, Y. Ivanov, G. Kostov, and S.Z.B. Thracian University Faculty of Veterinary Medicine (2000). Preparation of diagnostic agents-ingredients for ELISA and use of double sandwich procedures for detection of antigens and antibodies in avian influenza A (grippe). Bulgarian Journal of Veterinary Medicine 3(4): 163-170.  ISSN: 1311-1477.

            Abstract:  Type-specific antigens from horioallantoic membranes (HAM) and allanto-amnionic fluids (AAF) of chicken embryos (CE), infected with a referent avian influenza virus strain (AIV) subtype H2, as well as corresponding hyperimmune rabbit anf guinea pig sera were prepared. The latter, being highly specific and with a high sensitivity, were used as ingredients in an indirect double-sandwich ELISA procedure for detection of type-specific antigen of AIV and antibodies against it in a blockade double sandwich ELISA procedure. The results of blockade ELISA, applied to 916 hen sera from different farms from different regions of the country and to 11 sera from wild birds revealed no antibodies against AIV. Seven hundried and sixty eight of these sera where parallely studied in agar gel immunodiffusion (AGID) test and the results were negative as well. The studies, performed in the period 1993-1998 for isolation of AIV in Ce from viscera of 212 carcasses from 13 domestic and wild avian species, gave negative results.

            Descriptors:  avian influenza virus, antigens, antibodies, serotypes, ELISA, immunodiffusion tests,  biological differences, immunoenzyme techniques, immunological factors, immunological techniques, immunoprecipitation tests, influenza virus, orthomyxoviridae, viruses.

Hadjiev, G., G. Kostov, I. Chenchev, V. Bumbarov, Y. Ivanov, and S.Z.B. Thracian University Faculty of Veterinary Medicine (2000). Preparation of diagnosticums from referent avian influenza A virus (grippe) strains and attempts for the detection of the disease in Bulgaria. Bulgarian Journal of Veterinary Medicine 3(2-3): 81-88.  ISSN: 1311-1477.

            Abstract:  Inactivated antigens from the allantoamnionic fluids (AAF) and chorioallantoic membranes (CAM) of chicken embryos (CE) infected with the avian influenza A virus (grippe) were prepared. Referent viral strains from the subtypes H-2, H-5, H-6 and H-8 were used for that purpose. The titres for the 50% endpoint infection dose (EID50) of strains for HE were within the range 5.52-8.26 lg/ml and their haemagglutination titers - from 1:256 to 1:512. The antigens were predominantly used for screening studies of avian sera with the tests: agar gel immunodiffusion (AGID), haemagglutination inhibition (HI), complement fixation reaction (CFR) (with informative purpose) and the indirect immunofluorescence reaction (IIFR). With the AGID test, positive seroreagents were detected among samples from 2 farms. Using RHI in a previous period in other 2 farms, there were positive samples against the H-5 subtype.

            Descriptors:  avian influenza virus, antigens, immune serum, immunodiffusion tests, hemagglutination tests, complement fixation tests, immunofluorescence, agglutination tests, immunological factors, immunological techniques, immunoprecipitation tests, influenza virus, orthomyxoviridae, viruses.

Harley, V.R., P.J. Hudson, B.E. Coupar, P.W. Selleck, H. Westbury, and D.B. Boyle (1990). Vaccinia virus expression and sequence of an avian influenza nucleoprotein gene: potential use in diagnosis. Archives of Virology 113(1-2): 133-41.  ISSN: 0304-8608.

            NAL Call Number:  448.3 Ar23

            Abstract:  The nucleoprotein (NP) gene from avian influenza strain A/Shearwater/Aust/1/72 (H6N5) was cloned, sequenced, and expressed in vaccinia virus for the production of potent sera in immunised rabbits. The NP gene is 1565 bp and shares greater than 95% amino acid sequence identity with other NPs of the avian subtype. The recombinant NP expressed by vaccinia virus comigrated with endogenous A/Shearwater/Aust/1/72 NP by Western blot analysis. Polyclonal rabbit sera raised against recombinant NP was evaluated in an antigen capture ELISA system as a potential diagnostic tool for the detection of avian influenza. All type A strains, comprising several HA and NA subtypes, but not type B nor other avian viruses, were detected.

            Descriptors:  fowl plague diagnosis, genes viral, influenza A virus avian genetics, nucleoproteins genetics, vaccinia virus genetics, viral core proteins, viral proteins genetics, amino acid sequence, antibodies, viral immunology, base sequence, blotting, southern, cloning, molecular, DNA, viral, enzyme linked immunosorbent assay, avian immunology, molecular sequence data, nucleoproteins immunology, predictive value of tests, thymidine kinase genetics, vaccinia virus immunology, viral proteins immunology.

Horimoto, T. and Y. Kawaoka (1995). Direct reverse transcriptase PCR to determine virulence potential of influenza A viruses in birds. Journal of Clinical Microbiology 33(3): 748-751.  ISSN: 0095-1137.

            NAL Call Number:  QR46.J6

            Abstract:  A reverse transcriptase PCR (RT-PCR) was used for rapid determination of the hemagglutinin (HA) cleavage site sequence, a marker for the virulence potential of avian influenza viruses. When applied to specimens from chickens experimentally infected with either a virulent or an avirulent virus, RT-PCR uniformly detected the HA gene, even in specimens that were negative for virus by standard testing in eggs. This technique, combined with sequencing of the HA cleavage site, offers a rapid and sensitive way to assess the virulence potential of avian influenza viruses. Early detection of field isolates with virulence-associated structural motifs at the HA cleavage site would allow better control of influenza among large poultry populations.

            Descriptors:  chickens, avian influenza virus, pathogenicity, PCR, experimental infection, in vivo experimentation, agglutinins, genes, biological properties, birds, cell structure, chromosomes, disease transmission, domestic animals, domesticated birds, experimentation, Galliformes, infection, influenza virus, livestock, microbial properties, nucleus, orthomyxoviridae, pathogenesis, poultry,  proteins, useful animals, viruses, hemagglutinins.

Huang ShuJian  (1999). The diagnosis and control of avian influenza. Poultry Husbandry and Diseases Control (8): 8-10.

            Descriptors:  disease control, diagnosis, avian influenza virus, China.

Jin, M., G. Wang, R. Zhang, S. Zhao, H. Li, Y. Tan, and H. Chen (2004). Development of enzyme-linked immunosorbent assay with nucleoprotein as antigen for detection of antibodies to avian influenza virus. Avian Diseases 48(4): 870-8.  ISSN: 0005-2086.

            NAL Call Number:  41.8 Av5

            Abstract:  During the avian influenza outbreak of 2003-04 in Southeast Asia, two avian influenza viruses (AIV), one of H5N1 subtype and the other H9N2 subtype, were isolated and identified from local farms. The nudeoprotein (NP) gene of the H5N1 AI isolate was cloned, and the segment encoding amino acid 47-384, which covers its major antigenic domains, was subcloned and expressed in E. coli. Subsequently, the NP (47-384) expression product was purified and used as the diagnostic antigen to develop a NP-based type-specific indirect enzyme-linked immunosorbent assay (ELISA) for detecting antibodies to AI from chicken sera. The ELISA is shown to be specific for AIV and does not cross-react with chicken sera that has antibodies to other avian viruses. The NP(47-384)-ELISA was compared with a hemagglutination inhibition test and a commercial AIV ELISA kit in evaluating 150 sera samples from experimentally AIV-infected or vaccinated specific-pathogen-free (SPF) chickens. Our NP(47-384)-ELISA was more sensitive than the two tests and showed an 82% agreement ratio with the HI test and an 80.67% agreement ratio with the commercial kit. The NP(47-384)-ELISA and the commercial AIV ELISA were used to evaluate 448 field sera samples from diseased chickens or vaccinated chickens during the 2003-04 AI outbreak in China. The two ELISA tests had a 95% agreement ratio. We conclude that the NP(47-384)-ELISA developed in our laboratory was specific and sensitive and it has great application potential in China's long-term prevention and control of AI.

            Descriptors:  antibodies, viral blood, enzyme linked immunosorbent assay methods, influenza A virus, avian isolation and purification, nucleoproteins immunology, viral proteins immunology, amino acid sequence, chick embryo, chickens, avian immunology, avian influenza diagnosis, molecular sequence data, nucleoproteins chemistry, reagent kits, diagnostic, reproducibility of results, specific pathogen free organisms, viral proteins chemistry.

Jover, A., R. Manvell, R. Jackson, A. Medrano, A. Pages, and C. Artigas (2004). Screening for avian influenza: do it now! World Poultry 20(3): 26-27.  ISSN: 1388-3119.

            NAL Call Number:  SF481.M54

            Descriptors:  antibody testing, disease control, disease prevention, disease transmission, ELISA, public health, screening, zoonoses, avian influenza virus.

Kaleta, E.F., H. Will, E. Bernius, W. Kruse, and A.L. Bolte (1998). Zum serologischen Nachweis virusbedingter Infektionen bei der Hausgans ( Anser anser dom.). [The serologic detection of virus-induced infections in the domestic goose (Anser anser dom.)]. Tierarztliche Praxis. Ausgabe G, Grosstiere Nutztiere 26(4): 234-8.  ISSN: 1434-1220.

            NAL Call Number:  SF603.V43

            Abstract:  The most important virus-induced diseases associated with heavy losses in the domestic goose are Derzsy's disease which is caused by a goose parvovirus and duck plague (duck viral enteritis) which is caused by an avian herpesvirus. Both diseases still occur but can be prevented by timely vaccinations. Antibodies against Influenza A viruses of the subtypes H1, H5, and H7 as well as against avian paramyxoviruses of the serogroups 4, 6, and 8, respectively, were not detected in any of the examined sera. However, antibodies against paramyxovirus type 1 were detected in sera of one source. Haemagglutination inhibition or neutralizing antibodies against avian adenoviruses (EDS76 virus and goose adenovirus of the serotypes 1, 2, and 3) were quite often detected. Based on the present knowledge their pathogenic potential is minor. Neutralizing antibodies against a reovirus originating from Muscovy ducks and against a chicken reovirus (strain U Con S 1133) were quite frequently detected. In 35 of 564 examined geese sera hepatitis B virus was found.

            Descriptors:  antibodies, viral blood, geese, poultry diseases diagnosis, virus diseases veterinary, aviadenovirus immunology, avulavirus immunology, hepatitis B virus, duck immunology, hepatitis virus, duck immunology, influenza A virus avian immunology, parvovirus immunology, poultry diseases prevention and control, reoviridae immunology, virus diseases diagnosis, virus diseases prevention and control.

Kodihalli, S., V. Sivanandan, D.A. Halvorson, K.V. Nagaraja, and M.C. Kumar (1993). Antigen-capture ELISA for rapid diagnosis of avian influenza virus in commercial turkey flocks. Journal of Veterinary Diagnostic Investigation, Official Publication of the American Association of Veterinary Laboratory Diagnosticians, Inc 5(3): 438-40.  ISSN: 1040-6387.

            NAL Call Number:  SF774.J68

            Descriptors:  enzyme linked immunosorbent assay veterinary, fowl plague diagnosis, poultry diseases diagnosis, turkeys microbiology, antigens, viral analysis, cloaca microbiology, enzyme linked immunosorbent assay methods, fowl plague microbiology, influenza A virus avian isolation and purification, poultry diseases microbiology, sensitivity and specificity, trachea microbiology.

Kodihalli, S., V. Sivanandan, K.V. Nagaraja, S.M. Goyal, and D.A. Halvorson (1993). Antigen-capture enzyme immunoassay for detection of avian influenza virus in turkeys. American Journal of Veterinary Research 54(9): 1385-1390.  ISSN: 0002-9645.

            NAL Call Number:  41.8 Am3A

            Abstract:  A double-antibody sandwich ELISA (DAS-ELISA) was developed for detection of avian influenza virus (AIV) antigen. A monoclonal antibody to the viral nucleoprotein (NP) was used to coat the ELISA plates. A direct DAS-ELISA and an indirect DAS-ELISA were evaluated. In the direct DAS-ELISA, monoclonal antibody to the AIV NP conjugated with horseradish peroxidase was used. The direct DAS-ELISA was evaluated for its sensitivity to detect purified NP; this procedure detected as little as 0.1 ng. In the indirect DAS-ELISA, rabbit NP antibody and horseradish peroxidase-conjugated goat anti-rabbit immunoglobin were used as primary and secondary antibodies, respectively. The indirect DAS-ELISA was evaluated for its ability to detect the AIV antigen in tracheal and cloacal specimens from turkeys inoculated with AIV. Results of indirect DAS-ELISA were compared with those of conventional virus isolation. Percentage agreement between indirect DAS-ELISA and virus isolation in AIV-positive samples was found to be 76.1% and, in AIV-negative samples, it was found to be 82.1%. These results indicate that the DAS-ELISA might be a viable alternative to virus isolation because of its rapidity, compared with virus isolation.

            Descriptors:  turkeys, ELISA, avian influenza virus, antigens, monoclonal antibodies, animal viruses, proteins, antibodies, birds, Galliformes, immunoenzyme techniques, immunological factors, immunological techniques, immunology, influenza virus, viruses, viral nucleoproteins, viral antigens.

Kodihalli, S. (1993). Diagnosis and Control of Avian Influenza Virus Infection in Turkeys, p. viii, 148 leaves, ill.

            Descriptors:  avian influenza, diagnosis, control, turkeys.

Lagata, J.R., V. Sivanandan, A.S. Abraham, and J.A. Newman (1985). Monoclonal antibodies to hemagglutinin of avian influenza virus. Abstracts of Papers Presented at the Annual Meeting of the Conference of Research Workers in Animal Diseases. 66(Abstract 327): 60.

            NAL Call Number:  SF605.C59

            Descriptors:  hemagglutinins, avian influenza virus, diagnosis, monoclonal antibodies, turkeys, ducks.

Lagutkin, N.A. (1991). Aspects of the laboratory diagnosis of avian influenza and Newcastle disease. Veterinariia (3): 29-32.  ISSN: 0042-4846.

            NAL Call Number:  41.8 V6426

            Descriptors:  laboratory diagnosis, Newcastle disease, avian influenza virus.

Lamichhane, C.M. and L. Kirkegaard Jr. (1997). ELISA for the detection of antibodies to avian influenza type A virus in chicken sera. Proceedings of the Western Poultry Diseases Conference 46: 81.

            NAL Call Number:  SF995.W4

            Descriptors:  ELISA, detection, antibodies, avian influenza, chicken, sera, type A virus.

Lamichhane, C.M., D.E. Swayne, M. Blankfard, B. Erickson, and J. Beck (1996). Elisa for the detection of antibody to avian influenza type A virus in chicken and turkey serum. Proceedings of the Western Poultry Diseases Conference 45: 56-57.

            NAL Call Number:  SF995.W4

            Descriptors:  chickens, turkeys, avian influenza virus, birds, domestic animals, domesticated birds, Galliformes, influenza virus, livestock, orthomyxoviridae, poultry, useful animals, viruses.

Landgraf, J.G., J.E. Pearson, and D.A.n.p. Senne (1984). Laboratory diagnosis of avian influenza. Proceedings of the Western Poultry Diseases Conference 33: 3-4.

            NAL Call Number:  SF995.W4

            Descriptors:  laboratory diagnosis, avian influenza virus.

Lau, L.T., J. Banks, R. Aherne, I.H. Brown, N. Dillon, R.A. Collins, K.Y. Chan, Y.W. Fung, J. Xing, and A.C. Yu (2004). Nucleic acid sequence-based amplification methods to detect avian influenza virus. Biochemical and Biophysical Research Communications 313(2): 336-42.  ISSN: 0006-291X.

            NAL Call Number:  442.8 B5236

            Abstract:  Infection of poultry with highly pathogenic avian influenza virus (AIV) can be devastating in terms of flock morbidity and mortality, economic loss, and social disruption. The causative agent is confined to certain isolates of influenza A virus subtypes H5 and H7. Due to the potential of direct transfer of avian influenza to humans, continued research into rapid diagnostic tests for influenza is therefore necessary. A nucleic acid sequence-based amplification (NASBA) method was developed to detect a portion of the haemagglutinin gene of avian influenza A virus subtypes H5 and H7 irrespective of lineage. A further NASBA assay, based on the matrix gene, was able to detect examples of all known subtypes (H1-H15) of avian influenza virus. The entire nucleic acid isolation, amplification, and detection procedure was completed within 6h. The dynamic range of the three AIV assays was five to seven orders of magnitude. The assays were sensitive and highly specific, with no cross-reactivity to phylogenetically or clinically relevant viruses. The results of the three AIV NASBA assays correlated with those obtained by viral culture in embryonated fowl's eggs.

            Descriptors:  influenza A virus, genetics, isolation and purification, self sustained sequence replication methods, base sequence, birds, chick embryo, DNA primers genetics, DNA probes genetics, diagnosis, virology, sensitivity and specificity, species specificity, swine.

Lee, C.W. and D.L. Suarez (2004). Application of real-time RT-PCR for the quantitation and competitive replication study of H5 and H7 subtype avian influenza virus. Journal of Virological Methods 119(2): 151-8.  ISSN: 0166-0934.

            NAL Call Number:  QR355.J6

            Abstract:  Avian influenza (AI) viruses are endemic in wild birds and if transmitted to poultry can cause serious economic losses. In the study of AI, the quantitation of virus shed from infected birds is valuable in pathogenesis studies and to determine the effectiveness of vaccines, and is performed routinely by cultivation of virus containing samples using embryonating chicken eggs (ECE) and expressed by 50% egg infectious dose (EID(50)). Although, this assay is accurate and is the standard test for infectious virus titration, the method is laborious, requires a large number of ECE, and takes at least 7 days to determine results. In this study, a one-tube hydrolysis fluorescent probe based real-time RT-PCR (RRT-PCR) was applied for the quantitation of AI virus and compared with conventional virus titration method. A strong positive correlation was observed between the amount of RNA determined by quantitative RRT-PCR and the EID(50)s determined by conventional methods. This RRT-PCR test was further applied in the study of competitive replication of co-infected H5 and H7 subtype viruses in chickens. Using hemagglutinin subtype specific probes, we were able to determine the amount of individual subtype virus, which could not have easily been done with conventional methods. This RRT-PCR based quantitation of AI virus, which is specific, sensitive, easy to perform, and rapid, will be useful for virological, pathogenesis, and protection studies.

            Descriptors:  influenza A virus, avian physiology, avian classification, avian genetics, avian isolation and purification, poultry, sensitivity and specificity, poultry diseases virology, reverse transcriptase polymerase chain reaction methods, virus replication, fluorescent dyes, hemagglutinin glycoproteins, influenza virus analysis.

Levi, R., T. Beeor Tzahar, and R. Arnon (1995). Microculture virus titration--a simple colourimetric assay for influenza virus titration. Journal of Virological Methods 52(1-2): 55-64.  ISSN: 0166-0934.

            NAL Call Number:  QR355.J6

            Abstract:  Influenza antigens can be detected by several well established methods. However, when it is important to determine the titre of infective virions, a bioassay should be employed. The standard and the most widely used tests for influenza infectivity are titration carried out in embryonated hen eggs, or the plaque assay employing tissue culture techniques. A simple colourimetric assay for influenza virus detection and titration is described. Samples of allantoic fluid or mice lung homogenates were used to infect MDCK cultures in microplate wells. After an incubation period, the tetrazolium (MTT) colourimetric assay was used to determine cell viability, and when compared to untreated culture control enabled the detection and titration of several influenza strains. When samples were assayed simultaneously in embryonated eggs and by the MCVT method, good correlation in determined titres was obtained. The availability of an additional method for influenza titration allows more flexibility in the choice of titration method according to the specific needs of the study. Furthermore, this method lends itself to full automatization. Similar procedures should also be applicable to titration of other cytopathic viruses.

            Descriptors:  influenza virology, influenza A virus avian isolation and purification, human isolation and purification, cell line, chick embryo, colorimetry methods, dogs, avian physiology, human physiology, kidney, lung virology, mice, mice inbred BALB c, species specificity, virology methods, virus replication.

Lin, J.T. (2004). [Strategies in the diagnosis and treatment of patients with avian influenza]. Zhonghua Yi Xue Za Zhi 84(5): 355-6.  ISSN: 0376-2491.

            Descriptors:  influenza, avian influenza diagnosis, avian influenza drug therapy, acetamides therapeutic use, antiviral agents therapeutic use, birds virology, avian influenza A virus drug effects, avian influenza virology, sialic acids therapeutic use.

Lin, J.A. (1995). The technique of enzyme-linked immunosorbent assay on serology of avian influenza virus. Journal of the Chinese Society of Veterinary Science 21(3): 146-151.  ISSN: 0253-9179.

            NAL Call Number:  SF604.C54

            Abstract:  Enzyme-linked immunosorbent assay (ELISA) is widely used as a technique for serological diagnosis or seroepidemiological survey on various poultry diseases. Because avian influenza virus (AIV) seems to be getting severely threatened to economic animals in Taiwan, it is necessary to take a screening test for vast serum samples from animals. By the screening technique, it becomes to be possible to investigate the epidemiology of AIV infections. In this study, we are trying to develop the technique of AIV purification and ELISA to provide a model for laboratory work. The viral fluid was purified by different ultracentrifugation and sedimentation through a sucrose gradient (10-50% sucrose). ELISA was carried out in 96-well microplates coated with purified AIV antigens. Conjugated rabbit-anti-chicken peroxidase and substrate of o-Phenylenediamine dihydrochloride (OPD) were used in this study. The results of Box Titration of ELISA showed that the most proper concentration of coating antigen, conjugate and OPD were 1.0 mu g/well, 3,000X dilution and 0.4 mg/mL, respectively. Sera of 16X dilution were used. The high specificity of ELISA were conducted by using 40 AVI negative sera from specific-pathogen-free chickens, 5 positive sera of AIV, 5 positive sera of chicken anemia agent (CAA), 12 positive sera of Marek's disease virus (MDV), totally 62 tested sera.

            Descriptors:  enzymology, immune system, infection, microbiology, pathology, veterinary medicine, diagnosis.

Lin XiangMei, Zhao ZengLian, and Chen WanFang (2001). Detection of avian influenza virus antigen in tissues of chicken infected with A/duck/Nanjing/21/95. Chinese Journal of Veterinary Science 21(5): 466-470.  ISSN: 1005-4545.

            NAL Call Number:  SF604.C58

            Descriptors:  avian influenza virus, isolation, kidneys,  strains, chickens, China.

Lin XiangMei, Zhao ZengLian, Tian GuoBin, Chen WanFang, Chen PuYan, and Yu KangZhen (1998). Comparison of sensitivity among indirect ELISA, haemagglutinin inhibition (HI) and AGP tests for detecting antibodies against avian influenza A virus in chickens. Chinese Journal of Veterinary Science 18(5): 454-456.

            NAL Call Number:  SF604.C58

            Descriptors:  antibodies, ELISA, immunodiagnosis, influenza virus A and B, chickens.

Loza Rubio, E., F. Diosdado Vargas, A. Hernandez Magdaleno, V.M. Banda Ruiz, A. Morilla Gonzalez, and J. Garcia Garcia (1997). Diagnostico serologico de influenza aviar por medio de una tecnica de microimunodifusion (MIDG) en agar. (Nota de investigacion). [Serological diagnosis of avian influenza, using a microimmuno-diffusion test in agar][Research note]. Tecnica Pecuaria En Mexico (Mexico) 35(3): 165-169.  ISSN: 0040-1889.

            NAL Call Number:  49 T222

            Abstract:  The purpose of this study was to develop a microimmunodiffusion test in gel (MIDG), to detect antibodies against type A of avian influenza virus in chicken serum. An antigen was prepared from chorioallantonic membranes infected with the avian influenza virus strain (A/Ck/Puebla/14585-622/94H5N2). Microimmunodiffussion test in gel (MIDG) was compared with immunodiffusion test in agar (IDA), and Hemagglutination inhibition test (HI) which is the reference technique. Seventy five serum samples were obtained from specific pathogen free chicken, which resulted negative by the three assays. 163 serum samples were obtained from vaccinated poultry with HI titers from 1:10 to 1:1280. The sensitivity of these 2 agar gel precipitin tests was relatively low (43%) in comparison with HI. It was found that sensitivity of MIDG was identical to IDG (98%), and both were positives from HI titers of 1:320 and above. We conclude that IDA and MIDG are very useful in detecting infected poultry with virus that produce Hi titers from 1:320 to 1:1280. Both, can be used in laboratories with a reduced infrastructure, besides, MIDG has the advantage of making possible the analysis a great quantity of samples at low cost.

            Descriptors:  broiler chickens, avian influenza virus, immunodiagnosis, birds, chickens, diagnosis, domestic animals, Galliformes, immunological techniques, influenza virus, livestock, meat animals, orthomyxoviridae, poultry, useful animals, viruses.

Lu, H. (2003). A longitudinal study of a novel dot-enzyme-linked immunosorbent assay for detection of avian influenza virus. Avian Diseases 47(2): 361-369.  ISSN: 0005-2086.

            NAL Call Number:  41.8 Av5

            Abstract:  A monoclonal antibody (MAb)-based dot-enzyme-linked immunosorbent assay (ELISA) has been developed that detected the epitopes specifically associated with avian influenza virus (AIV). The dot-ELISA detected the antigens of AIV directly from clinical and field specimens. Data obtained from experimentally AIV-infected specific-pathogen-free chickens and also the 2001/02 AIV outbreak of serotype H7N2 positive flocks in Pennsylvania indicated that the mean sensitivity (Se) of the dot-ELISA ranged between 45% and 68% and the mean specificity (Sp), between 85% and 90%. The values were derived from various clinical and field specimens when compared with virus isolation with embryonating chicken eggs. On routine AIV surveillance samples, the dot-ELISA achieved a 92%-100% Sp on the basis of testing over 1500 AIV surveillance samples that were confirmed negative by virus isolation. The dot-ELISA detected AIV antigens with a 5-mul allantoic fluid sample that contained a concentration of 0.4 hemagglutinating units. Furthermore, the dot-ELISA retained its specificity for AIV because no cross-reactions were obtained with various other avian viruses. The findings in this study indicated that the dot-ELISA was highly sensitive and specific and comparable with the commercial Directigen(R) test in the detection of AIV obtained from clinical and field specimens.

            Descriptors:  immune system, infection, avian influenza, diagnosis, infectious disease, respiratory system disease, viral disease, monoclonal antibody based dot ELISA, mab dot ELISA, immunologic techniques, laboratory techniques.

Malik, Y.S., D.P. Patnayak, and S.M. Goyal (2004). Detection of three avian respiratory viruses by single-tube multiplex reverse transcription-polymerase chain reaction assay. Journal of Veterinary Diagnostic Investigation, Official Publication of the American Association of Veterinary Laboratory Diagnosticians, Inc 16(3): 244-8.  ISSN: 1040-6387.

            NAL Call Number:  SF774.J68

            Abstract:  Acute respiratory tract infections are leading causes of morbidity in poultry farms throughout the world. Avian pneumovirus (APV), avian influenza virus (AIV), and Newcastle disease virus (NDV) have been recognized as the most important pathogens of both chicken and turkeys. Single-virus reverse transcription-polymerase chain reaction (sRT-PCR) assays are used extensively to detect these viruses in clinical samples. This study reports the development and evaluation of a single-tube multiplex RT-PCR (mRT-PCR) assay for simultaneous and specific detection of APV, AIV, and NDV. Specific primers for each virus were selected that amplified products of predicted sizes from each virus in the mRT-PCR as well as in the sRT-PCR assays (438, 218, and 532 bp for APV, AIV, and NDV, respectively). The sensitivity and specificity of mRT-PCR assay were compared with those of the sRT-PCR. The mRT-PCR assay was as sensitive as the sRT-PCR assays because virus detection limits were similar in both assays. The detection limits of mRT-PCR assay were 10(0.5) tissue culture infective dose (50%) (TCID50)/ml, 10(1.2) TCID50/ml, and 10(0.7) TCID50/ml for APV, AIV, and NDV, respectively. Overall, there was an excellent correlation between mRT-PCR and sRT-PCR assays. No product amplification was obtained with nucleic acid from infectious bronchitis virus and reovirus using these primer sets. In summary, mRT-PCR assay holds potential to be an economical and rapid diagnostic method for the simultaneous detection of 3 avian respiratory viruses in chickens and turkeys.

            Descriptors:  influenza A virus, avian growth and development, metapneumovirus growth and development, Newcastle disease virus growth and development, poultry diseases virology, respiratory tract infections veterinary, reverse transcriptase polymerase chain reaction veterinary, turkeys, avian genetics, metapneumovirus genetics, Newcastle disease virus genetics, RNA, viral chemistry, viral genetics, respiratory tract infections diagnosis, respiratory tract infections virology, reverse transcriptase polymerase chain reaction methods, sensitivity and specificity.

Manvell, R. (1998). Diagnosi di laboratorio dell' Influenza aviare. [Laboratory diagnosis of avian influenza]. [Italian Society of Poultry Pathology. Meeting on World and Italian situation of avian influenza]. Legnaro, Padua (Italy). 7 Apr 1998. Selezione Veterinaria (Italy) (12): 952-955.

            NAL Call Number:  241.71 B75

            Descriptors:  chickens, laboratory diagnosis, avian influenza virus, viroses, animal diseases, immunological techniques, pathogenicity, identification, enzyme inhibitors, nucleotide sequence, antimetabolites, biological properties, birds, diagnosis, domestic animals, Galliformes, genomes, infectious diseases, influenza virus, livestock, microbial properties, orthomyxoviridae, poultry, useful animals, viruses.

McNulty, M.S., J.B. McFerran, J.B. McFerran (ed.), and M.S. McNulty (ed.) (1986). Avian influenza: diagnosis and vaccination. Current Topics in Veterinary Medicine and Animal Science - Acute Virus Infections of Poultry 37: 36-44.

            NAL Call Number:  SF600.C82

            Descriptors:  avian influenza virus, diagnosis, vaccination.

Merino, R., L.J. Gutierrez, V.R. Tejeda, and J.A. Quintana (2002). Detection of antibodies against avian influenza virus in egg yolk, using HI, ELISA and AGID tests.  Poultry Science (USA) 81(Suppl. 1): 163.  ISSN: 0032-5791.

            NAL Call Number:  47.8 Am33P

            Descriptors:  infection, methods and techniques, veterinary medicine, avian influenza, respiratory system disease, viral disease, ELISA diagnostic method, agar gel immunodiffusion test diagnostic method, hemagglutination inhibition test diagnostic method, meeting abstract.

Min, S.L., C.C. Poa, H.S. Jui, H.L. Long, and H.K. Shieh (1999). Identification and differentiation of avian influenza and Newcastle disease viruses by a multiplex reverse transcription-polymerase chain reaction. Journal of the Chinese Society of Veterinary Science 25(3): 191-198.  ISSN: 0253-9179.

            NAL Call Number:  SF604.C54

            Abstract:  Avian influenza and Newcastle disease share many common clinical symptoms. A method for rapid identification and differentiation of these two diseases comes to be necessary. A pair of primers was designed based on the nucleotide sequences of the highly conserved region of nucleoprotein of avian influenza virus. By reverse transcription-polymerase chain reaction (RT-PCR), this pair of primers could amply a 330 bp fragment from 13 reference strains (H1-H13) and 77 field isolates of avian influenza virus with a high sensitivity and specificity. A multiplex RT-PCR procedure was then developed by using the above AIV primers and primers for the detection of Newcastle disease virus (NDV) developed in the previous study. This multiplex RT-PCR procedure could detect both AIV and NDV in a single reaction tube, and could differentiate vaccine strains from field isolates of NDV. Since the one tube multiplex RT-PCR could significantly reduce the time and cost of AIV and NDV, it is suitable for rapid identification and differentiation of AIV and NDV.

            Descriptors:  molecular genetics, multiplex reverse transcriptase polymerase chain reaction (multiplex RT PCR) DNA amplification method, polymerase chain reaction, nucleotide sequencing sequencing method.

Mitchell, B.W. and M. Brugh (1982). Comparison of electrocardiograms of chickens infected with viscerotropic velogenic Newcastle disease virus and virulent avian influenza virus. American Journal of Veterinary Research 43(12): 2274-8.  ISSN: 0002-9645.

            NAL Call Number:  41.8 Am3A

            Abstract:  Electrocardiograms of chickens infected with viscerotropic velogenic Newcastle disease virus (NDV) or virulent avian influenza virus (AIV) were characterized and compared. The ECG were monitored by radiotelemetry and were recorded twice daily before virus infection and during the course of the infection. Thirteen lead II intervals, segments, and amplitudes were measured and analyzed. The ECG of NDV-infected chickens were characterized by lengthened (P less than or equal to 0.05) ST segments and increased (P less than or equal to 0.05) P amplitudes. The ECG of AIV-infected chickens were characterized by lengthened (P less than or equal to 0.05) RS intervals, ST segments, TP intervals, and PR segments and by increased (P less than or equal to 0.05) P amplitudes. The TP intervals and PR segments of ECG of AIV-infected chickens were significantly (P less than or equal to 0.05) longer than those of NDV-infected chickens. The pronounced conduction delays indicated in the ECG of AIV-infected chickens may have diagnostic importance.

            Descriptors:  chickens, fowl plague physiopathology, heart physiopathology, Newcastle disease physiopathology, electrocardiography veterinary, influenza A virus avian pathogenicity, Newcastle disease virus pathogenicity, specific pathogen free organisms, virulence.

Moreno, A. and R. Ruiz (1983). La influenza aviar. Epizootiologia, diagnostico y control. [Epidemiology, diagnosis and control of avian influenza - a review]. Revista Avicultura, Cuba 27(3): 89-108.

            NAL Call Number:  SF481.A9

            Descriptors:  reviews, avian influenza virus, epidemiology, diagnosis, control.

Munch, M., L.P. Nielsen, K.J. Handberg, and P.H. Jorgensen (2001). Detection and subtyping (H5 and H7) of avian type A influenza virus by reverse transcription-PCR and PCR-ELISA. Archives of Virology 146(1): 87-97.  ISSN: 0304-8608.

            NAL Call Number:  448.3 Ar23

            Abstract:  Avian influenza virus infections are a major cause of morbidity and rapid identification of the virus has important clinical, economical and epidemiological implications. We have developed a one-tube Reverse Transcriptase Polymerase Chain Reaction (RT-PCR) for the rapid diagnosis of avian influenza A. A panel of reference influenza strains from various hosts including avian species, human, swine and horse were evaluated in a one tube RT-PCR using primers designed for the amplification of a 218 bp fragment of the NP gene. The PCR products were detected by PCR-ELISA by use of an internal catching probe confirming the NP influenza A origin. The PCR-ELISA was about 100 times more sensitive than detection of PCR products by agarose gel electrophoresis. RT-PCR and detection by PCR-ELISA is comparable in sensitivity to virus propagation in eggs. We also designed primers for the detection of the influenza. A subtypes H5 and H7 shown to have pathogenic potential in poultry. The H5 primers cover the cleavage site of the HA gene and specifically amplify influenza A subtype H5. The H7 primers also cover the HA cleavage site and detected all H7 reference strains investigated. In addition, the H7 primers also amplified very weak and/or additional bands on an agarose gel from other subtypes. However, the H7 origin and the pathogenic potential defined by the presence or absence of basic amino acids at the cleavage site can be determined by sequencing of the PCR product. As far as we know this is the first demonstration of RT-PCR detection on a panel of H7 strains using only one primer set.

            Descriptors:  influenza A virus avian isolation and purification, polymerase chain reaction methods, birds, DNA primers genetics, enzyme linked immunosorbent assay, fowl plague virology, hemagglutinins viral genetics, horses, avian classification, avian genetics, nucleoproteins analysis, nucleoproteins genetics, RNA viral analysis, reverse transcriptase polymerase chain reaction, sensitivity and specificity, swine.

Naeem, K., M. Naurin, S. Rashid, and S. Bano (2003). Seroprevalence of avian influenza virus and its relationship with increased mortality and decreased egg production. Avian Pathology 32(3): 285-289.  ISSN: 0307-9457.

            NAL Call Number:  SF995.A1A9

            Descriptors:  antibody testing, disease prevalence, disease surveys, seroprevalence, ELISA, egg production, poultry, mortality, avian influenza virus, Pakistan.

Ninomiya, A. (1998). Detection of antibodies in the sera specific to the hemagglutinin of avian influenza viruses. Japanese Journal of Veterinary Research 46(2-3): 135-136.  ISSN: 0047-1917.

            NAL Call Number:  41.8 V6446

            Descriptors:  infection, veterinary medicine, influenza virus infection, viral disease, thesis, dissertation.

Oskolkov, V.S., K.h. Kreimer Yu, and A.A. Ibragimov (1974). [Some problems of epidemiology and diagnosis of avian influenza]. Veterinariia (4): 63-65.

            NAL Call Number:  41.8 V6426

            Descriptors:  avian influenza, epidemiology, diagnosis, problems, poultry.

Palfi, V., L. Tekes, and J. Tanyi (1997). Improvement of diagnostic methods for infectious diseases. Magyar Allatorvosok Lapja 119(11): 686-692.  ISSN: 0025-004X.

            NAL Call Number:  41.8 V644

            Abstract:  Diagnostic methods to be used in case of notifiable infectious diseases have been regulated partly by regulations and partly by guidelines to be considered obligatory. The use of internationally accepted standard methods is especially important because the veterinary authorities issue different transport papers, certificates based on the results of these investigations. Only with a few exceptions, the diagnosis of infectious diseases in the list A of Office International des Epizooties is the  responsibility of central veterinary laboratories in each country. The laboratory procedures are based on the demonstration of virus its antigen or nucleic acid in the animal tissues, isolation of virus, as well as demonstration of antibodies developing in the blood serum by different methods. ELISA and virus isolation are used for the demonstration of foot-and-mouth disease (FMD) virus. Serum antibodies are demonstrated by virus neutralization or ELISA tests. In case of swine vesicular disease  (SVD) the samples are tested parallel also for the presence of FMD virus. The SVD virus can be isolated in cell cultures of porcine origin and can not be isolated in those of bovine origin. Serum antibodies are demonstrated by virus neutralization and ELISA tests. Immunofluorescent investigation of tissue samples and virus isolation methods are used for the diagnosis of classical swine fever (CSF). Monoclonal antibodies are used to differentiate among CSF, BVD virus and Suvac vaccine virus  strains. Different ELISA and virus neutralization test are used for the demonstration of serum antibodies. In case of a suspected African Swine Fever case, haemadsorption and immunofluorescent tests are used for the demonstration of the virus and also pigs are inoculated with the suspected material. ELISA is used for the serological diagnosis of EBL. Cultivation of avian influenza viruses is carried out in embryonated eggs. The type of virus is determined by immuno-diffusion test and the type  of haemagglutinin and neuraminidase - and occasionally - the virulence have also been determined. Haemagglutination inhibition test is used for the demonstration of serum antibodies. Practically the same methods are used for the diagnosis of Newcastle disease.

            Descriptors:  animal husbandry, infection, methods and techniques, infectious disease, swine fever, virus neutralization test diagnostic method, ELISA diagnostic method.

Pang, Y., H. Wang, T. Girshick, Z. Xie, and M.I. Khan (2002). Development and application of a multiplex polymerase chain reaction for avian respiratory agents. Avian Diseases 46(3): 691-9.  ISSN: 0005-2086.

            NAL Call Number:  41.8 Av5

            Abstract:  A multiplex polymerase chain reaction (PCR) was developed and optimized to simultaneously detect 6 avian respiratory pathogens. Six sets of specific oligonucleotide primers for infectious bronchitis virus (IBV), avian influenza virus (AIV), infectious laryngotracheitis virus (ILTV), Newcastle disease virus (NDV), Mycoplasma gallisepticum (MG), and Mycoplasma synoviae (MS) were used respectively in the test. With the use of agarose gel electrophoresis for detection of the PCR-amplified DNA products, the sensitivity of detection was between 10 pg for IBV, AIV, MG, and ILTV and 100 pg for NDV and MS after 35 cycles of PCR. Similar sensitivity of these primers was achieved with chickens experimentally infected with respiratory pathogens. In experimental infections, the multiplex PCR was able to detect all the infected chickens in each group at I and 2 wk postinfection as compared with serologic tests at 2 wk postinfection that confirmed the presence of specific antibodies. The multiplex PCR was also able to detect and differentiate coinfections with two or more pathogens. No specific DNA amplification for respiratory avian pathogens was observed among noninoculated birds kept separately as a negative control group.

            Descriptors:  bird diseases diagnosis, Mycoplasma infections veterinary, polymerase chain reaction veterinary, virus diseases veterinary, bird diseases microbiology, bird diseases virology, birds, chick embryo, chickens, DNA, bacterial isolation and purification, DNA, viral isolation and purification, diagnosis, differential,  electrophoresis, agar gel veterinary, Mycoplasma genetics, Mycoplasma isolation and purification, Mycoplasma infections diagnosis, polymerase chain reaction methods, RNA viral isolation and purification, reverse transcriptase polymerase chain reaction methods, reverse transcriptase polymerase chain reaction veterinary, sensitivity and specificity, species specificity, specific pathogen free organisms, virus diseases diagnosis.

Pang YaoShan, Xie ZhiXun, and Khan M. I. (2001). Establishement of two-temperature multiplex polymerase chain reaction (PCR) for detecting six pathogens causing respiratory diseases in chicks at the same time. Chinese Journal of Veterinary Science and Technology 31(5): 3-6.  ISSN: 1000-6419.

            Descriptors:  avian influenza virus, infectious bronchitis virus, infectious laryngotracheitis virus, Mycoplasma gallisepticum, Mycoplasma synoviae, Newcastle disease virus, diagnostic techniques, experimental infections, polymerase chain reaction, poultry, chicks.

Patolsky, F., G. Zheng, O. Hayden, M. Lakadamyali, X. Zhuang, and C.M. Lieber (2004). Electrical detection of single viruses. Proceedings of the National Academy of Sciences of the United States of America 101(39): 14017-22.  ISSN: 0027-8424.

            NAL Call Number:  500 N21P

            Abstract:  We report direct, real-time electrical detection of single virus particles with high selectivity by using nanowire field effect transistors. Measurements made with nanowire arrays modified with antibodies for influenza A showed discrete conductance changes characteristic of binding and unbinding in the presence of influenza A but not paramyxovirus or adenovirus. Simultaneous electrical and optical measurements using fluorescently labeled influenza A were used to demonstrate conclusively that the conductance changes correspond to binding/unbinding of single viruses at the surface of nanowire devices. pH-dependent studies further show that the detection mechanism is caused by a field effect, and that the nanowire devices can be used to determine rapidly isoelectric points and variations in receptor-virus binding kinetics for different conditions. Lastly, studies of nanowire devices modified with antibodies specific for either influenza or adenovirus show that multiple viruses can be selectively detected in parallel. The possibility of large-scale integration of these nanowire devices suggests potential for simultaneous detection of a large number of distinct viral threats at the single virus level.

            Descriptors:  influenza A virus, avian isolation and purification, nanotechnology methods, paramyxoviridae isolation and purification, birds, electric conductivity, immunochemistry, avian chemistry, avian immunology, avian metabolism, microscopy, electron, transmission, microscopy, fluorescence, nanotechnology instrumentation, paramyxoviridae chemistry, paramyxoviridae immunology, paramyxoviridae metabolism, silicon chemistry.

Payungporn, S., P. Phakdeewirot, S. Chutinimitkul, A. Theamboonlers, J. Keawcharoen, K. Oraveerakul, A. Amonsin, and Y. Poovorawan (2004). Single-step multiplex reverse transcription-polymerase chain reaction (RT-PCR) for influenza A virus subtype H5N1 detection. Viral Immunology 17(4): 588-93.  ISSN: 0882-8245.

            Abstract:  Influenza A virus subtype H5N1 causes a rapidly fatal systemic disease in domestic poultry and spreads directly from poultry to humans. The aim of this study was to develop a rapid, cost-saving and effective method for influenza A virus subtype H5N1 detection. The selected primer set was used in single-step RT-PCR for simultaneous detection in multiplex format of the 276-, 189-, and 131-bp fragments, corresponding to sequences specific for M, H5 and N1. The amplified DNA fragments were clearly separated by agarose gel electrophoresis. The sensitivity of this assay was about 10(3) copies/microL. Moreover, this method can be applied to detect not only avian but also human influenza A virus subtype H5N1. In conclusion, the highlights of this particular method are its rapidity and cost-effectiveness, thus rendering it feasible and attractive for large-scale screening at times of influenza A virus subtype H5N1 outbreak.

            Descriptors:  influenza virology, influenza A virus, avian isolation and purification, avian influenza virology, reverse transcriptase polymerase chain reaction methods, birds virology, chickens virology, influenza diagnosis, avian genetics, avian influenza diagnosis, sensitivity and specificity.

Pearson, J.E. and D.A. Senne. (1981). Avian influenza diagnostic procedures. In: Proceedings of the First International Symposium on Avian Influenza, Beltsville, Maryland, USA, p. 157-166.

            NAL Call Number: aSF995.6.I6I5 1981a

            Descriptors: avian influenza virus, tests, assays, diagnostic procedures, symposium.

Pearson, J.E., D.A. Senne, and D.A. Halvorson. ( 1986). Diagnosis of avian influenza in the United States. In: IVth International Symposium of Veterinary Laboratory Diagnosticians, p. 647-650.

            NAL Call Number: SF771.I5 1986

            Descriptors: avian influenza virus, diagnosis, turkeys, United States, symposium.

Perdue, M.L., P.O. Wainright, and M. Brugh (1990). Effects of chicken embryo age on time to death following infection by avian influenza viruses: implications for distinguishing highly pathogenic isolates. Virus Research 16(2): 137-52.  ISSN: 0168-1702.

            NAL Call Number:  QR375.V6

            Abstract:  When white leghorn (WL) chick embryos ranging in age from 8 to 13 days were inoculated with a variety of avian influenza virus (AIV) isolates, strain-specific differences in embryo mean death times (MDT) were observed. Non-highly pathogenic (nHP) strains killed 8 or 9 day-old embryos much more rapidly than 12 or 13 day-old embryos. Highly pathogenic (HP) strains, however, were less sensitive to embryo age resulting in similar MDTs in both older and younger embryos. These observations were consistent over a broad range of virus doses for both HP and nHP strains. When a HP derivative of H5N2 AIV was compared to its nHP parent, the derivative killed older embryos more rapidly than the parent virus, while MDTs in younger embryos were the same for both parent and derivative. The two strains further exhibited clear differences in the structure of their respective hemagglutinin, a previously described pathogenicity determinant for this virus. Thus it may be possible to readily demonstrate the HP phenotype in AIV strains based on MDT measurements in WL embryos.

            Descriptors:  chick embryo microbiology, orthomyxoviridae pathogenicity, orthomyxoviridae infections veterinary, poultry diseases mortality, antibodies, monoclonal immunology, hemagglutinin glycoproteins, influenza virus, hemagglutinins viral immunology, orthomyxoviridae infections mortality, specific pathogen free organisms, time factors, viral envelope proteins immunology.

Pourbakhsh, S.A., F. Moosakhani, and M. Kianizadeh (1999). Standardization of ELISA for detection of avian influenza virus antibodies. Archives of Razi Institute (50): 19-28.  ISSN: 0365-3439.

            NAL Call Number:  QR189.A73

            Descriptors:  ELISA, hemagglutination inhibition test, immunodiagnosis, antibody testing, diagnostic techniques, poultry, influenza virus.

Purchase, H.G. (1989). Practical application of nucleic acid techniques to avian disease problems. Avian Diseases 33(4): 609-14.  ISSN: 0005-2086.

            NAL Call Number:  41.8 Av5

            Abstract:  A workshop in which 17 practicing scientists participated was intended to address primarily people who use or could use biotechnology in their work and was confined to five techniques. Endonuclease fingerprinting and mapping involved cleaving nucleic acid with a specific restriction enzyme and separating the nucleic acid fragments by electrophoresis. Field and vaccine isolates of Pasteurella multocida could be distinguished; Salmonella enteritidis could be divided into three groups; chlamydia could be grouped into seven groups; and vaccinia, quail pox, and fowl pox could be clearly distinguished. Preparation of nucleic acid probes involved producing large amounts of labeled oligonucleotides, usually of unknown sequence. Successful probes had been made for infectious bursal disease virus, avian influenza virus, Newcastle disease virus, and infectious bronchitis virus. In Southern, Northern, and dot blotting, either DNA or RNA fragments were placed on or transferred to a solid substrate and probed. The procedure was able to detect infectious bursal disease virus, infectious bronchitis virus, Mycoplasma gallisepticum, and Marek's disease virus. In situ hybridization involved applying a labeled probe to frozen or fixed sections or to intact cells. In Polymerase chain reaction, two primers, some distance apart, were annealed to a denatured target DNA. Repeated cycles of DNA synthesis with a thermostable polymerase, denaturing, and reannealing resulted in great amplification of a rare sequence. After 30 cycles, a rare gene sequence could be amplified more than 10(6) times. It was used successfully to detect minute quantities of influenza virus and infectious bursal disease virus, and the process was used to facilitate DNA sequencing of coccidiosis gene segments.

            Descriptors:  poultry diseases diagnosis, blotting, northern, blotting, Southern, DNA restriction enzymes genetics, nucleic acid hybridization, nucleic acid probes, peptide mapping, polymerase chain reaction.

Rada, B. and T. Hanusovska (1987). Rapid method for the detection of synergism in combinations of antiviral substances. Acta Virologica 31(2): 126-37.  ISSN: 0001-723X.

            NAL Call Number:  448.3 AC85

            Abstract:  The plaque inhibition method was modified in order to evaluate the effectiveness of various combinations of antiviral substances. One substance (A) diffuses from the centre of cell culture, the other (B) is incorporated into the agar overlay at subinhibitory concentration. The inhibitory effect of the combination (A + B) is demonstrated by the increase in size of the inhibitory zone in comparison with the control inhibitory zone produced by the substance A alone. The ratio of the diameter of the inhibitory zone with substance combination (A + B) to the diameter of single drug control zone (substance A) serves as index DI (degree of interaction). Quantitative evaluation of the degree of potentiation using isobolograms showed that DI greater than 1.5 indicate a synergistic effect of the respective combinations. This inexpensive method can serve for rapid selection of suitable combinations out of number of substances. Model experiments were performed with combinations of selected inhibitors of virus replication.

            Descriptors:  antiviral agents pharmacology, influenza A virus avian drug effects, vaccinia virus drug effects, chick embryo, cycloheximide pharmacology, drug synergism, drug therapy, combination, microbial sensitivity tests, models, biological, plaque assay, ribavirin pharmacology, rimantadine pharmacology.

Ramos, V.J.A., P.F. Del, M. Kiupel, S.D. Fitzgerald, A.J. Bermudez, G.C. Johnson, and M.A. Miller (2002). Diagnostic immunohistochemistry of equine and avian infectious diseases. Journal of Histotechnology 25(4): 185-198.  ISSN: 0147-8885.

            Abstract:  This article is the second part of a series of review articles dedicated to immunohistochemical detection of infectious agents in domestic animals and covers infectious diseases of horses and birds. Immunohistochemistry is heavily used in these species for certain infectious diseases and until recently was the only quick and reliable diagnostic technique for some diseases (e.g., West Nile virus). A discussion of the immunohistochemical detection of the following infectious diseases of horses (Borna disease, eastern equine encephalitis virus, equine herpesvirus 1, equine protozoal myelitis, equine viral arteritis, leishmaniasis, viral papillomatosis, rabies, and West Nile fever) and birds (avian adenovirus, avian influenza virus, avian pneumovirus, budgerigar fledgling disease, chlamydiosis, Newcastle disease virus, Tyzzer's disease, and West Nile fever) is presented. In addition, references of selected equine and avian infectious diseases in which immunohistochemistry has been used and immunohistochemical protocols from the authors' laboratories are tabulated.

            Descriptors:  infection, veterinary medicine, borna disease, diagnosis, infectious disease, viral disease, Newcastle disease, infectious disease, viral disease, west nile fever, infectious disease, nervous system disease, viral disease, abortion, reproductive system disease, female, infectious disease, infectious disease, formalin fixation histology and cytology techniques, laboratory techniques, immunohistochemistry clinical techniques, diagnostic techniques, immunologic techniques, laboratory techniques, paraffin embedding histology and cytology techniques, laboratory techniques.

Rowe, T., R.A. Abernathy, J. Hu Primmer, W.W. Thompson, X. Lu, W. Lim, K. Fukuda, N.J. Cox, and J.M. Katz (1999). Detection of antibody to avian influenza A (H5N1) virus in human serum by using a combination of serologic assays. Journal of Clinical Microbiology 37(4): 937-43.  ISSN: 0095-1137.

            NAL Call Number:  QR46.J6

            Abstract:  From May to December 1997, 18 cases of mild to severe respiratory illness caused by avian influenza A (H5N1) viruses were identified in Hong Kong. The emergence of an avian virus in the human population prompted an epidemiological investigation to determine the extent of human-to-human transmission of the virus and risk factors associated with infection. The hemagglutination inhibition (HI) assay, the standard method for serologic detection of influenza virus infection in humans, has been shown to be less sensitive for the detection of antibodies induced by avian influenza viruses. Therefore, we developed a more sensitive microneutralization assay to detect antibodies to avian influenza in humans. Direct comparison of an HI assay and the microneutralization assay demonstrated that the latter was substantially more sensitive in detecting human antibodies to H5N1 virus in infected individuals. An H5-specific indirect enzyme-linked immunosorbent assay (ELISA) was also established to test children's sera. The sensitivity and specificity of the microneutralization assay were compared with those of an H5-specific indirect ELISA. When combined with a confirmatory H5-specific Western blot test, the specificities of both assays were improved. Maximum sensitivity (80%) and specificity (96%) for the detection of anti-H5 antibody in adults aged 18 to 59 years were achieved by using the microneutralization assay combined with Western blotting. Maximum sensitivity (100%) and specificity (100%) in detecting anti-H5 antibody in sera obtained from children less than 15 years of age were achieved by using ELISA combined with Western blotting. This new test algorithm is being used for the seroepidemiologic investigations of the avian H5N1 influenza outbreak.

            Descriptors:  antibodies, viral blood, influenza A virus avian immunology, serologic tests methods, adolescent, adult, blotting, western methods, blotting, western statistics and numerical data, child, preschool, cross reactions, enzyme linked immunosorbent assay methods, enzyme linked immunosorbent assay statistics and numerical data, hemagglutination inhibition tests methods, hemagglutination inhibition tests statistics and numerical data, Hong Kong epidemiology, influenza epidemiology, influenza immunology, influenza transmission, avian classification, avian pathogenicity, middle aged, neutralization tests methods, neutralization tests statistics and numerical data, sensitivity and specificity, seroepidemiologic studies, serologic tests statistics and numerical data.

Russi Cahill, J.C., M.C. Mogdasy, R.E. Somma Moreira, and M.H. de Peluffo (1975). Counterimmunoelectrophoresis with influenza antigens. I. Use of avian plague virus to detect type-specific antibodies to influenza A in human sera. Journal of Infectious Diseases 131(1): 64-6.  ISSN: 0022-1899.

            NAL Call Number:  448.8 J821

            Abstract:  Avian plague virus was used as antigen in a counterimmunoelectrophoresis technique. This virus was selected because it detects only type-specific influenza A antibodies in human sera, avoiding the possible interference of other antigens with anodic migration. The results with reference sera, as well as the correlation of positive sera found by counterimmunoelectrophoresis and complement fixation with the proposed antigen, in the absence of other types of antibodies to fowl plague virus antigen, support the conclusion that the counterimmunoelectrophoresis technique reveals type-specific antibodies. The test is more sensitive than immunodiffusion but less sensitive than complement fixation. Its sensitivity, simplicity, and rapidity make it suitable for serologic surveys of human influenza A.

            Descriptors:  antibodies, viral analysis, antigens, viral, immunoelectrophoresis, influenza immunology, influenza A virus avian immunology, antibody specificity, chick embryo, complement fixation tests, guinea pigs immunology,  immune sera, immunodiffusion.

Sala, G., P. Cordioli, M.A. Moreno, M. Tollis, E. Brocchi, A. Piccirillo, and A. Lavazza (2003). ELISA test for the detection of influenza H7 antibodies in avian sera. Avian Diseases 47(Special Issue): 1057-1059.  ISSN: 0005-2086.

            NAL Call Number:  41.8 Av5

            Abstract:  Using a monoclonal antibody (MAb) specific for the H7 influenza surface glycoproteins, a serological enzyme-linked immunosorbent assay (ELISA) test has been developed. This MAb was made using the low-pathogenicity (LP) avian influenza (AI) strain (BS2676/99) isolated in Italy during a recent outbreak. The test is able to detect H7 antibodies in avian sera. The H7 ELISA has a 99% concordance of results with the classical hemagglutination inhibition (HI) test.

            Descriptors:  immune system, infection, ELISA, immunologic techniques, laboratory techniques, hemagglutination inhibition test.

Samadieh, B. (1971). Pathogenicity, transmissibility and diagnosis of two strains of avian influenza-A viruses in turkeys. Dissertation Abstracts International, B 31(9): 5440.

            NAL Call Number:  Z5055.U49D53

            Descriptors:  avian influenza A virus, diagnosis, pathogenicity, transmissibility, turkeys.

Samadieh, B., R. Kargar Moaakhar, and M. Afnan ( 1975). Demonstration of avian influenza-A virus in Iran by immunodiffusion technique. Avian Diseases 19(4): 689-91.  ISSN: 0005-2086.

            NAL Call Number:  41.8 Av5

            Abstract:  A total of 1000 chicken serum samples (CSS) and 235 turkey serum samples (TSS) were tested by an immunodiffusion procedure against soluble antigen (S-antigen) prepared from avian influenza-A virus (AIAV), T/Calif/5142/66. None of the CSS tested developed any precipitin line, whereas 8.9% of the TSS tested developed well-defined precipitin lines against S-antigen. This observation confirmed the presence of AIAV in Iran.

            Descriptors:  influenza A virus avian immunology, chickens immunology, fowl plague immunology, Iran, turkeys immunology.

Samame, H.A. and R.A. Bankowski (1977). Application of the egg-bit technique in poultry disease research. Avian Diseases 21(4): 623-629.  ISSN: 0005-2086.

            NAL Call Number:  41.8 Av5

            Descriptors:  avian influenza virus, cell culture techniques, egg-bit technique, virology, eggs, poultry, disease research.

Sangret, M., O.J. Vrtiak, and B. Kapitancik (1972). Use of DEAE-cellulose in testing disinfectants. Acta Virologica 16(5): 444.  ISSN: 0001-723X.

            NAL Call Number:  448.3 AC85

            Descriptors:  antiviral agents pharmacology, cellulose, disinfectants pharmacology, adsorption, body fluids microbiology, chick embryo, drug resistance, microbial, fetal membranes, hemagglutination tests, influenza A virus avian drug effects, avian isolation and purification, methods, sodium chloride, sodium hydroxide.

Selleck, P.W., S.L. Lowther, G.M. Russell, and P.T. Hooper (2003). Rapid diagnosis of highly pathogenic avian influenza using pancreatic impression smears. Avian Diseases 47(Special Issue): 1190-1195.  ISSN: 0005-2086.

            NAL Call Number:  41.8 Av5

            Abstract:  The 1985 outbreak of high-pathogenicity avian influenza (HPAI) in Victoria, Australia, took 5 days to confirm by standard laboratory tests, during which time infected chickens continued excreting virus, thus creating the opportunity for transmission to other farms. An immunofluorescence test for the detection of viral antigen in tissue impression smears was evaluated as a rapid diagnostic test for HPAI virus infections of poultry. Several test configurations were compared for background reactions and strength of fluorescence, with the optimum combination found to be an influenza A group-specific monoclonal antibody, detected by an anti-mouse fluorescein isothiocyanate conjugate. Immunohistochemical examination of tissues from chickens experimentally infected with low-pathogenicity and HPAI viruses identified the pancreas as the organ most consistently containing high concentrations of HPAI viral antigen. This test has since been used in Australia in the rapid laboratory confirmation of three avian influenza outbreaks and in showing that numerous other suspect cases were not caused by avian influenza.

            Descriptors:  epidemiology, infection, avian influenza, diagnosis, infectious disease, transmission, viral disease, immunofluorescence, immunologic techniques, laboratory techniques, immunohistochemistry, pancreatic impression smears, clinical techniques, diagnostic techniques, pathogenicity.

Semenova, N.P., E.N. Kantorovich Prokudina, I.A. Miasnikova, and O.N. Berezina (1978). Ispol'zovanie metoda bliashek dlia vyiavleniia fenomena autointerferentsii u virusa grippa. [Use of the plaque method to reveal the phenomenon of autointerference in influenza virus]. Voprosy Virusologii (6): 709-13.  ISSN: 0507-4088.

            NAL Call Number:  448.8 P942

            Abstract:  The paper describes a simple and convenient method for qualitative and quantitative evaluation of the capacity of influenza virus for autointerference consisting in the lack or considerable reduction of the cytolytic effect of the virus under agar overlay at a high multiplicity of infection. Some experimental and theoretical arguments assuming the role of defective interfering particles in the formation of the observed phenomenon. It is assumed that the detection of autointerference under agar may be used as an additional criterion for detection of non-plaque-forming strains of influenza virus, tentative determination of their interfering capacity as well as for the establishment of biological relationships of viruses.

            Descriptors:  influenza A virus physiology, viral interference, defective viruses, influenza A virus avian, plaque assay.

Shafer, A.L., J.B. Katz, and K.A. Eernisse (1998). Development and validation of a competitive enzyme-linked immunosorbent assay for detection of type A influenza antibodies in avian sera. Avian Diseases 42(1): 28-34.  ISSN: 0005-2086.

            NAL Call Number:  41.8 Av5

            Abstract:  Serologic screening of avian sera for group-specific antibodies to type A influenza is currently accomplished by using the avian influenza (AI) agar gel immunodiffusion (AGID) test. A competitive enzyme-linked immunosorbent assay (CELISA) was developed using a baculovirus vector, Autographa californica nuclear polyhedrosis virus, expressing the nucleoprotein (NP) gene of A/Ann Arbor/6/60 influenza virus. The recombinant NP was obtained by inoculation of Spodoptera frugiperda  (Sf9) insect cells or Trichoplusia ni insect larvae with the recombinant baculovirus. A hybridoma cell line producing monoclonal antibody against influenza virus A nucleoprotein was used to generate mouse ascitic fluid for the CELISA. The nucleoprotein and the monoclonal antibody were used without further purification in a CELISA for detection of avian-origin serum antibodies to type A influenza. The AI AGID and CELISA tests were compared for sensitivity and specificity using 1651 experimental and reference antisera. Samples discrepant in AGID and CELISA test results were further evaluated by the AI indirect fluorescent antibody (IFA), hemagglutination-inhibition (HI), and neuraminidase-inhibition (NI) tests. The results demonstrated a high degree of correlation between the AGID and CELISA test results, with the IFA, HI, and NI tests offering additional support of CELISA test specificity. The CELISA is a rapid, economical, sensitive, and specific serodiagnostic method for screening large numbers of avian sera for antibodies to avian influenza virus.

            Descriptors:  birds, avian influenza virus, ELISA, antibodies, body fluids, immunology, immunodiagnosis, testing, recombinant antigens, microbial proteins, animal viruses, gene expression, cell culture, monoclonal antibodies, evaluation, immunological techniques, antibodies, antigens, body parts, culture techniques, diagnosis, immunoenzyme techniques, immunological factors, immunological techniques, in vitro culture, influenza virus, orthomyxoviridae, viruses, serum, serology, screening, cell lines, comparisons.

Shamseddini, M., M. Vasfi Marandi, S.A. Pourbakhsh, M. Gharagozlo,  M. Bahmani-Nejad, and P. Khazraee-Nia (2002). The use of indirect immunoperoxidase assay in diagnosis of type A (H9N2) avian influenza virus antigen on frozen tissue sections. Archives of Razi Institute (53): 11-21.  ISSN: 0365-3439.

            NAL Call Number:  QR189.A73

            Descriptors:  animal tissues, staining techniques, immunoperoxidase technique, diagnosis, experimental infection, immune response, avian influenza virus, broilers, poultry.

Shan, S., L.S. Ko, R.A. Collins, Z. Wu, J. Chen, K.Y. Chan, J. Xing, L.T. Lau, and A.C.H. Yu (2003). Comparison of nucleic acid-based detection of avian influenza H5N1 with virus isolation. Biochemical and Biophysical Research Communications 302(2): 377-383.  ISSN: 0006-291X.

            NAL Call Number:  442.8 B5236

            Abstract:  Nucleic acid sequence-based amplification with electrochemiluminescent detection (NASBA/ECL) of avian influenza virus was compared with viral culture in embryonated chicken eggs. Virus was isolated from blood or anal swabs of chickens artificially infected with highly pathogenic avian influenza A/Chicken/Hong Kong/1000/97 (H5N1). Viral nucleic acid was detected in blood samples by NASBA/ECL immediately prior to death, whilst nucleic acid extracted from anal swabs was detected from the day following artificial infection until death. Thus, blood and/or anal swabs are a suitable source of material for the detection of avian influenza in dead birds, but anal swabs are more suitable for detection of viral genetic material in live birds. Dilution of a known viral standard was used to determine the limit of sensitivity for both NASBA/ECL and egg culture detection methods. The NASBA/ECL method was equivalent in sensitivity to egg culture. The NASBA/ECL results agreed with egg culture data in 71/94 (75.5%) tissue samples obtained from artificially infected birds.

            Descriptors:  infection, molecular genetics, avian influenza virus infection, viral disease, electrochemiluminescence laboratory techniques, nucleic acid sequence based amplification genetic techniques, laboratory techniques, virus isolation.

Shao HongXia, Qin Kun, Liu YeLong, and Jin WenJie (2002). Development of monoclonal antibody specific to haemagglutinins of subtype H5 avian influenza virus. Chinese Journal of Animal Quarantine 19(8): 21-23.  ISSN: 1005-944X.

            Descriptors:  avian influenza virus, diagnosis, hemagglutinins, monoclonal antibodies, poultry.

Sivanandan, V., D.A. Halvorson, and J.A. Newman (1988). Phagocytosis assay for avian influenza virus in turkeys. Proceedings of the Western Poultry Diseases Conference 51: 37.

            NAL Call Number:  SF995.W4

            Descriptors:  diagnostic techniques, avian influenza virus, turkeys, phagocytosis assay.

Sivanandan, V., C.K. Perera, K.V. Nagaraja, and D.A. Halvorson (1996). Avian influenza virus antibody detection by competitive ELISA. Proceedings of the Western Poultry Diseases Conference 45: 52.

            NAL Call Number:  SF995.W4

            Descriptors:  ELISA, avian influenza virus, immunoenzyme techniques, immunological techniques, influenza virus, orthomyxoviridae, viruses.

Smolenskii, V.I. and O.I. Sukharev (1974). Preparation of luminescent antibody against avian influenza virus, strain GP6. Sbornik Nauchnykh Trudov Moskovskaya Veterinarnaya Akademiya 73(2): 41-42.

            Descriptors:  avian influenza virus, mmunofluorescence, poultry, antibody.

Snyder, D.B., W.W. Marquardt, F.S. Yancey, and P.K. Savage (1985). An enzyme-linked immunosorbent assay for the detection of antibody against avian influenza virus. Avian Diseases 29(1): 136-44.  ISSN: 0005-2086.

            NAL Call Number:  41.8 Av5

            Abstract:  An enzyme-linked immunosorbent assay (ELISA) was developed for detecting antibody to type A avian influenza (AI) virus. The sensitivity and group specificity of the AI-ELISA were compared with those of the agar-gel-precipitin test (AGPT) and the hemagglutination-inhibition (HI) test under conditions of both controlled and field exposure. During the course of temporal experimental infection (0-76 days) of specific-pathogen-free (SPF) chickens with AI subtype Hav9N2, the AI-ELISA was able to detect specific AI antibody as early as 8 days postinoculation (PI), and it measured rising levels of antibody through 35 days PI, at which time the chickens were re-exposed to AI virus. Conversely, AGP tests were negative through 35 days PI, and HI tests began to detect low levels of AI antibody only at 21 days PI. Following a secondary infection at 35 days PI with the same AI subtype, all tests measured rising levels of AI-specific antibody (35-76 days PI). However, the AGP test was positive at only the 7- and 14-day samplings postsecondary immunization. Under field conditions, the AI-ELISA was able to detect serum AI antibody in flocks from which highly pathogenic AI was isolated, but the AGP tests of these sera were negative.

            Descriptors:  antibodies, viral analysis, chickens, fowl plague immunology, influenza A virus avian immunology, enzyme linked immunosorbent assay, hemagglutination inhibition tests veterinary, precipitin tests veterinary.

Soto, P.E., Z.A. Heneidi, A.D. Vazquez, M.E. Aranda, J. Gonzalez, P. Reyes, G.M. Gay, and J.G. Garcia (1996). Valoracion de la prueba de hi utilizada en el montoreo serologico de la influenza aviar. [Evaluation of the hemagglutination inhibition test for serologically monitoring of avian influenza]. Proceedings of the Western Poultry Diseases Conference 45: 53-56.

            NAL Call Number:  SF995.W4

            Descriptors:  hemagglutination inhibition test, evaluation, monitoring, avian influenza.

Spackman, E., D.A. Senne, L.L. Bulaga, T.J. Myers, M.L. Perdue, L.P. Garber, K. Lohman, L.T. Daum, and D.L. Suarez (2003). Development of real-time RT-PCR for the detection of avian influenza virus. Avian Diseases 47(Special Issue): 1079-1082.  ISSN: 0005-2086.

            NAL Call Number:  41.8 Av5

            Abstract:  A real-time reverse transcriptase/polymerase chain reaction (RRT-PCR) assay was developed using hydrolysis probes for the detection of avian influenza virus (AIV) and the H5 and H7 subtypes. The AIV specific primers and probes were directed to regions of the AIV matrix gene that are conserved among most type A influenza viruses. The H5 and H7 primers and probes are directed to H5 and H7 hemagglutinin gene regions that are conserved among North American avian influenza viruses. The sensitivity and specificity of this RRT-PCR assay was compared to virus isolation (VI) in chicken embryos with 1550 clinical swab samples from 109 live-bird markets (LBMs) in New York and New Jersey. RRT-PCR detected influenza in samples from 61 of 65 (93.8%) of the LBMs that were the sources of VI positive samples. Of the 58 markets that were positive for H7 influenza by hemagglutination inhibition assay, RRT-PCR detected H7 influenza in 56 markets (96.5%). Too few H5 positive samples were obtained to validate the H5 RRT-PCR assay in this study. Although RRT-PCR was less sensitive than VI on an individual sample basis, this study demonstrated that the AIV and H7 RRT-PCR assays are good tools for the rapid screening of flocks and LBMs.

            Descriptors:  infection, molecular genetics, avian influenza, infectious disease, respiratory system disease, viral disease, hemagglutination inhibition assay bioassay techniques, immunologic techniques, laboratory techniques, real time reverse transcriptase polymerase chain reaction, real time RT PCR, clinical techniques, diagnostic techniques, genetic techniques, viral isolation, diagnostic techniques, live bird markets.

Spackman, E., D.A. Senne, L.L. Bulaga, S. Trock, and D.L. Suarez (2003). Development of multiplex real-time RT-PCR as a diagnostic tool for avian influenza. Avian Diseases 47(Special Issue): 1087-1090.  ISSN: 0005-2086.

            NAL Call Number:  41.8 Av5

            Abstract:  A multiplex real-time reverse transcriptase-polymerase chain reaction (RRT-PCR) assay for the simultaneous detection of the H5 and H7 avian influenza hemagglutinin (HA) subtypes was developed with hydrolysis type probes labeled with the FAM (H5 probe) and ROX (H7 probe) reporter dyes. The sensitivity of the H5-H7 subtyping assay was determined, using in vitro transcribed RNA templates, to have a reproducible detection limit for H7 of approximately 104 HA gene copies and approximately 104-105 HA gene copies of H5. A direct comparison of H5-H7 multiplex RRT-PCR with hemagglutination inhibition (HI) was performed with 83 AI RRT-PCR and virus isolation positive tracheal and cloacal swab samples obtained from various avian species and environmental swabs from live-bird markets in New York and New Jersey. Both multiplex RRT-PCR and HI agreed on the subtype determination of 79 (95.2%) of the 83 samples, of which 77 were positive for H7 and two were determined to be non-H5/non-H7 subtypes. No samples were determined to be the H5 subtype by either assay.

            Descriptors:  immune system, infection, molecular genetics, avian influenza, infectious disease, respiratory system disease, viral disease, hemagglutination inhibition clinical techniques, diagnostic techniques, immunologic techniques, laboratory techniques, multiplex real time reverse transcriptase polymerase chain reaction, multiplex RT PCR, clinical techniques, genetic techniques.

Spackman, E., D.A. Senne, T.J. Myers, L.L. Bulaga, L.P. Garber, M.L. Perdue, K. Lohman, L.T. Daum, and D.L. Suarez (2002). Development of a real-time reverse transcriptase PCR assay for type A influenza virus and the avian H5 and H7 hemagglutinin subtypes. Journal of Clinical Microbiology 40(9): 3256-60.  ISSN: 0095-1137.

            NAL Call Number:  QR46.J6

            Abstract:  A real-time reverse transcriptase PCR (RRT-PCR) assay based on the avian influenza virus matrix gene was developed for the rapid detection of type A influenza virus. Additionally, H5 and H7 hemagglutinin subtype-specific probe sets were developed based on North American avian influenza virus sequences. The RRT-PCR assay utilizes a one-step RT-PCR protocol and fluorogenic hydrolysis type probes. The matrix gene RRT-PCR assay has a detection limit of 10 fg or approximately 1,000 copies of target RNA and can detect 0.1 50% egg infective dose of virus. The H5- and H7-specific probe sets each have a detection limit of 100 fg of target RNA or approximately 10(3) to 10(4) gene copies. The sensitivity and specificity of the real-time PCR assay were directly compared with those of the current standard for detection of influenza virus: virus isolation (VI) in embryonated chicken eggs and hemagglutinin subtyping by hemagglutination inhibition (HI) assay. The comparison was performed with 1,550 tracheal and cloacal swabs from various avian species and environmental swabs obtained from live-bird markets in New York and New Jersey. Influenza virus-specific RRT-PCR results correlated with VI results for 89% of the samples. The remaining samples were positive with only one detection method. Overall the sensitivity and specificity of the H7- and H5-specific RRT-PCR were similar to those of VI and HI.

            Descriptors:  fowl plague virology, hemagglutinin glycoproteins, influenza virus genetics, influenza A virus avian isolation and purification, poultry diseases virology, reverse transcriptase polymerase chain reaction, chick embryo, fluorescent dyes, hemagglutination inhibition tests, avian classification,  avian genetics, poultry, sensitivity and specificity.

Spackman, E. and D.L. Suarez (2005). Use of a novel virus inactivation method for a multicenter avian influenza real-time reverse transcriptase-polymerase chain reaction proficiency study. Journal of Veterinary Diagnostic Investigation, Official Publication of the American Association of Veterinary Laboratory Diagnosticians, Inc 17(1): 76-80 .  ISSN: 1040-6387.

            NAL Call Number:  SF774.J68

            Abstract:  Proficiency assessments are important elements in quality control for diagnostic laboratories. Traditionally, proficiency testing for polymerase chain reaction (PCR)-based assays has involved the use of clinical samples, samples "spiked" with live agents or DNA plasmids. Because of government regulations and biosecurity concerns, distribution of live high-consequence pathogens of livestock and poultry, such as avian influenza, is not possible, and DNA plasmids are not technically suitable for evaluating RNA virus detection. Therefore, a proficiency testing panel using whole avian influenza in a diluent containing a phenolic disinfectant that inactivates the virus while preserving the RNA for at least 8 weeks at -70 C was developed and used in a multicenter proficiency assessment for a type A influenza real-time reverse transcriptase (RT)-PCR test. The test, which was highly standardized, except for variation in the real-time RT-PCR equipment used, was shown to be highly reproducible by proficiency testing in 12 laboratories in the United States, Canada, and Hong Kong. Variation in cycle threshold values among 35 data sets and 490 samples was minimal (CV = 5.19%), and sample identifications were highly accurate (96.7% correct identifications) regardless of real-time PCR instrumentation.

            Descriptors:  disinfectants pharmacology, influenza A virus, avian isolation and purification, laboratories standards, reverse transcriptase polymerase chain reaction veterinary, virus inactivation drug effects, avian drug effects, quality control, RNA, viral drug effects, reproducibility of results, reverse transcriptase polymerase chain reaction standards.

Starick, E., O.A. Roemer, and O. Werner (2000). Type- and subtype-specific RT-PCR assays for avian influenza A viruses (AIV). Journal of Veterinary Medicine Series B 47(4): 295-301.  ISSN: 0931-1793.

            NAL Call Number:  41.8 Z52

            Abstract:  Reverse transcriptase (RT) PCR assays have been developed to improve the diagnosis of avian influenza A. RT-PCR using primers complementary to a conserved region of the matrix protein was assessed as being suitable for the detection of influenza A virus RNA from poultry as well as from pigs, horses and humans, regardless of the haemagglutinin (HA) and neuraminidase (NA) subtype. Therefore, this RT-PCR is a valuable tool to confirm the initial diagnosis of any influenza A infection. As a second approach, experiments were performed to identify the HA gene encoding the post-translational cleavage site of potentially highly pathogenic AIV isolates by RT-PCR. The principal aim was to design one universal primer pair for each virus subtype, H5 and H7, respectively, which allows the detection of all strain variants using only one consistent method. To realize this objective, it was necessary to develop 'wobble' primers. AIV RNAs from seven H5 and 11 H7 subtype viruses included in the investigations were specifically recognized by RT-PCR using these primers. This method therefore provides a rapid, subtype-specific diagnosis and subsequent sequencing of H5 and H7 avian influenza viruses.

            Descriptors:  animal husbandry, infection, methods and techniques, influenza A virus infection, avian, diagnosis, viral disease, reverse transcriptase polymerase chain reaction diagnostic method, polymerase chain reaction.

Starick, E. and O. Werner (2003). Detection of H7 avian influenza virus directly from poultry specimens.  Avian Diseases 47(Special Issue): 1187-1189.  ISSN: 0005-2086.

            NAL Call Number:  41.8 Av5

            Abstract:  Using clinical materials from experimentally infected poultry, we established an effective method for the preparation of viral RNA directly from tissue samples and eggs. Furthermore, our type A-specific matrix reverse transcription-polymerase chain reaction (RT-PCR) test was improved, and an H7 subtype-specific nested RT-PCR, which includes the hemagglutinin cleavage site, was designed. Both RT-PCR systems proved to be as sensitive as virus isolation. In addition, the labeled H7 HA-nested PCR primers were suitable for sequencing of the PCR products. The RT-PCR amplification of viral RNA and sequencing of the PCR product allows for the sensitive and rapid differentiation between low-pathogenic and highly pathogenic avian influenza viruses.

            Descriptors:  immune system, infection, molecular genetics, avian influenza, infectious disease, respiratory system disease, viral disease, H7 subtype, specific nester, reverse transcriptase polymerase chain reaction, clinical techniques, RT-PCR, diagnostic techniques, genetic techniques, laboratory techniques, genetic techniques, type A specific matrix.

Stephenson, I., J.M. Wood, K.G. Nicholson, and M.C. Zambon (2003). Sialic acid receptor specificity on erythrocytes affects detection of antibody to avian influenza haemagglutinin. Journal of Medical Virology 70(3): 391-398.  ISSN: 0146-6615.

            Abstract:  Haemagglutination-inhibition tests (HI) are used to detect increases in influenza antibody in serum. However, they are relatively insensitive for the detection of human antibody responses to avian haemagglutinin, even in the presence of high titres of neutralising antibody after confirmed infection or vaccination. Human influenza viruses bind preferentially sialic acid containing N-acetylneuraminic acid alpha2,6-galactose (SAalpha2,6Gal) linkages while avian and equine viruses bind preferentially those containing N-acetylneuraminic acid alpha2,3-galactose (SAalpha2,3Gal) linkages. Increasing the proportion of SAalpha2,3Gal linkages on the erythrocytes used, by enzymatic modification or change of species, improves the ability of erythrocytes to bind to avian influenza strains and thereby improves the sensitivity of detection of antibody to avian and equine HA in a range of mammalian and human sera using HI tests.

            Descriptors:  clinical chemistry, hematology, infection, methods and techniques, serology, veterinary medicine, hemagglutinin inhibition test clinical techniques, diagnostic techniques, immunologic techniques, laboratory techniques.

Suarez, D.L., E. Spackman, D.A. Senne, L. Bulaga, A.C. Welsch, and K. Froberg (2003). The effect of various disinfectants on detection of avian influenza virus by real time RT-PCR. Avian Diseases 47(Special Issue): 1091-1095.  ISSN: 0005-2086.

            NAL Call Number:  41.8 Av5

            Abstract:  An avian influenza (AI) real time reverse transcriptase-polymerase chain reaction (RRT-PCR) test was previously shown to be a rapid and sensitive method to identify AI virus-infected birds in live-bird markets (LBMs). The test can also be used to identify avian influenza virus (AIV) from environmental samples. Consequently, the use of RRT-PCR was being considered as a component of the influenza eradication program in the LBMs to assure that each market was properly cleaned and disinfected before allowing the markets to be restocked. However, the RRT-PCR test cannot differentiate between live and inactivated virus, particularly in environmental samples where the RRT-PCR test potentially could amplify virus that had been inactivated by commonly used disinfectants, resulting in a false positive test result. To determine whether this is a valid concern, a study was conducted in three New Jersey LBMs that were previously shown to be positive for the H7N2 AIV. Environmental samples were collected from all three markets following thorough cleaning and disinfection with a phenolic disinfectant. Influenza virus RNA was detected in at least one environmental sample from two of the three markets when tested by RRT-PCR; however, all samples were negative by virus isolation using the standard egg inoculation procedure. As a result of these findings, laboratory experiments were designed to evaluate several commonly used disinfectants for their ability to inactivate influenza as well as disrupt the RNA so that it could not be detected by the RRT-PCR test. Five disinfectants were tested: phenolic disinfectants (Tek-trol and one-stroke environ), a quaternary ammonia compound (Lysol no-rinse sanitizer), a peroxygen compound (Virkon-S), and sodium hypochlorite (household bleach). All five disinfectants were effective at inactivating AIV at the recommended concentrations, but AIV RNA in samples inactivated with phenolic and quaternary ammonia compounds could still be detected by RRT-PCR. The peroxygen and chlorine compounds were effective at some concentrations for both inactivating virus and preventing amplification by RRT-PCR. Therefore, the RRT-PCR test can potentially be used to assure proper cleaning and disinfection when certain disinfectants are used.

            Descriptors:  animal husbandry, infection, avian influenza, infectious disease, respiratory system disease, viral disease, environmental disinfection clinical techniques, therapeutic and prophylactic techniques, real time reverse transcriptase polymerase chain reaction clinical techniques, diagnostic techniques, genetic techniques, laboratory techniques, influenza eradication program, live bird markets.

Swayne, D.E. and H.D. Stone (1996). Valuation of inactivated H5 avian influenza virus and fowlpox virus recombinant vaccines and diagnostic test reagents: implications in avian influenza control and prevention strategies. Proceedings of the Western Poultry Diseases Conference 45: 35-38.

            NAL Call Number:  SF995.W4

            Descriptors:  avian influenza virus, vaccines, influenza virus, orthomyxoviridae, viruses.

Syurin, V.N., G.A. Ivanova, E.A. Krasnobaev and  B. Fomin Yu (1972). Laboratornaya Diagnostika Virusnykh Boleznei Zhivotnykh. [Laboratory Diagnosis of Viral Diseases of Animals], 416 p.

            Descriptors:  virology, laboratory diagnosis, farm animals, identification.

Tumpey, T.M., R. Alvarez, D.E. Swayne, and D.L. Suarez (2005). Diagnostic approach for differentiating infected from vaccinated poultry on the basis of antibodies to NS1, the nonstructural protein of influenza A virus. Journal of Clinical Microbiology 43(2): 676-83.  ISSN: 0095-1137.

            NAL Call Number:  QR46.J6

            Abstract:  Vaccination programs for the control of avian influenza (AI) in poultry have limitations due to the problem of differentiating between vaccinated and virus-infected birds. We have used NS1, the conserved nonstructural protein of influenza A virus, as a differential diagnostic marker for influenza virus infection. Experimentally infected poultry were evaluated for the ability to induce antibodies reactive to NS1 recombinant protein produced in Escherichia coli or to chemically synthesized NS1 peptides. Immune sera were obtained from chickens and turkeys inoculated with live AI virus, inactivated purified vaccines, or inactivated commercial vaccines. Seroconversion to positivity for antibodies to the NS1 protein was achieved in birds experimentally infected with multiple subtypes of influenza A virus, as determined by enzyme-linked immunosorbent assay (ELISA) and Western blot analysis. In contrast, animals inoculated with inactivated gradient-purified vaccines had no seroconversion to positivity for antibodies to the NS1 protein, and animals vaccinated with commercial vaccines had low, but detectable, levels of NS1 antibodies. The use of a second ELISA with diluted sera identified a diagnostic test that results in seropositivity for antibodies to the NS1 protein only in infected birds. For the field application phase of this study, serum samples were collected from vaccinated and infected poultry, diluted, and screened for anti-NS1 antibodies. Field sera from poultry that received commercial AI vaccines were found to possess antibodies against AI virus, as measured by the standard agar gel precipitin (AGP) test, but they were negative by the NS1 ELISA. Conversely, diluted field sera from AI-infected poultry were positive for both AGP and NS1 antibodies. These results demonstrate the potential benefit of a simple, specific ELISA for anti-NS1 antibodies that may have diagnostic value for the poultry industries.

            Descriptors:  animals, viral blood antibodies, chickens, avian influenza A virus classification, avian influenza A virus immunology, avian influenza diagnosis, avian influenza immunology, avian influenza prevention and control, avian influenza virology, poultry, poultry diseases diagnosis, poultry diseases prevention and control, poultry diseases virology, non-U.S. Government research support, non-U.S. Government P.H.S. research support, turkeys, vaccination, viral nonstructural proteins chemical synthesis, viral nonstructural proteins immunology, viral vaccines administration and dosage, viral antibodies, influenza virus INS1 protein, viral nonstructural proteins, viral vaccines.

Van Deusen, R.A., V.S. Hinshaw, D.A. Senne, and D. Pellacani (1983). Micro neuraminidase-inhibition assay for classification of influenza A virus neuraminidases. Avian Diseases 27(3): 745-750.  ISSN: 0005-2086.

            NAL Call Number:  41.8 Av5

            Descriptors:  avian influenza virus, inhibition assay, classification.

Velek, K. and V. Leathers (1999). Evaluation of a highly sensitive and specific ELISA for detection of antibodies to avian influenza virus. Proceedings of the Western Poultry Diseases Conference 48: 140-141.

            NAL Call Number:  SF995.W4

            Descriptors:  avian influenza virus, ELISA, detection.

Volokhov, D.V., N.A. Vlasov, and I.M. Surgucheva (1999). The bi-sitespecific monoclonal antibodies in avian influenza A virus nucleoprotein ELISA. Sel'Skokhozyaistvennaya Biologiya 0(4): 91-99.  ISSN: 0131-6397.

            NAL Call Number:  S13.S44

            Abstract:  The data on production of quadromas secreting bi-sitespecific monoclonal immunoglobulins to avian influenza A virus nucleoprotein and horseradsh peroxidase are presented. The monoclonal antibodies application in immunoassay was shown to be possible by development of test-system for avian influenza laboratory diagnostics.

            Descriptors:  immune system, immunoassay diagnostic method, laboratory diagnostics diagnostic method, nucleoprotein, ELISA, analytical method.

Wang XiuRong, Deng GuoHua, Yu KangZhen, Qiao ChuanLing, Liu LiLing, Chen HuaLan, and Jiang YongPing (2004). [Detection and subtyping of avian influenza virus using DNA microarray hybridization]. Animal Biotechnology Bulletin 9(1): 356-357.  ISSN: 1014-8469.

            Descriptors:  avian influenza virus, characterization, diagnosis, diagnostic techniques, DNA, genetic analysis, microarray, hybridization, subtyping, poultry, fowl.

Wang ZeLin, L.i. Wang, Yao HuiXia, Ma ZhongBin, and Song YunQing (2004). Rapid differentiation of the virulence of Newcastle disease virus isolates by RT-PCR. Chinese Journal of Veterinary Science 24(4): 317-320.  ISSN: 1005-4545.

            NAL Call Number:  SF604.C58

            Descriptors:  Newcastle disease, amino acid sequences, avian infectious bursitis, bronchitis, death, diagnostic techniques, embryos, pathogenicity, polymerase chain reaction, reverse transcription, allantoic fluid, avian influenza virus, poultry, fowl.

Waschke, K. and H. Staber (1976). Stich -- Plaque -- Test, ein okonomisches Verfahren zur quantitativen Virusbestimmung [Stich-plaque test--an economic method for quantitative determination of viruses]. Journal of Hygiene, Epidemiology, Microbiology, and Immunology 21(3): 332-40.  ISSN: 0022-1732.

            NAL Call Number:  448.8 J826

            Abstract:  An economic method for quantitative assay of viruses is presented. In this "canule stick-plaque test" (German abbreviation SPT) samples of viruses, geometrically diluted and taken up by a canule, are inoculated by sticking into monolayer cell cultures overlayed with agar medium. A plaquelike CPE detectable by neutral red staining develops in the area of the inoculation. The frequency of this CPE formation depends on the concentration of viruses in the inoculated dilution. This dose-response allows calculation of the ID 50. In this way it is possible to carry out titration involving 6 dilutions and 10 inoculations per dilution using 3 common Petri dishes (6 cm in diameter), only. The sensitivity, accuracy, and reproductibility of this method are described and discussed.

            Descriptors:  plaque assay methods, viruses isolation and purification, chick embryo, cytopathogenic effect, viral, herpesvirus 1, suid isolation and purification, influenza A virus avian isolation and purification, sindbis virus isolation and purification, tissue culture.

Watanabe, S., H. Shimizu, C. Kawakami, and M. Imai (1999). [Evaluation of an optical immunoassay test for rapid detection of influenza A and B viral antigen]. Kansenshogaku Zasshi Journal of the Japanese Association for Infectious Diseases 73(12): 1199-204.  ISSN: 0387-5911.

            Abstract:  An optical immunoassay test, FLU OIA (BIOSTAR, USA), was evaluated for the diagnosis of influenza viral infection. The reactivity of the FLU OIA test was evaluated using 42 influenza strains (25 human influenza A, 12 human influenza B, 2 swine influenza A and 3 avian influenza A strains). The FLU OIA test showed positive results for all influenza strains. There was no evidence that cross-reactivity occurred with non-influenza viruses. The detection limit of the FLU OIA test was found to be 3.0-6.5 x 10(4) pfu/assay for human influenza A and B strains. The sensitivity and specificity of the FLU OIA test compared to isolation in cell culture was 89.7% and 76.0% for testing of 54 nasopharyngeal aspirate specimens. The FLU OIA test is rapid and easy for the detection of influenza A and B viral antigen and provide a valuable tool for the rapid diagnosis of influenza viral infection.

            Descriptors:  antigens, viral analysis, influenza A virus immunology, influenza B virus immunology, evaluation studies, immunoassay methods.

Werner, O. (1987). Beitrag zur Verbesserung der Diagnostik von Influenza-A-Virus-infektionen des Geflugels. [Improving the diagnosis of influenza A virus infections in poultry]. Archiv Fur Experimentelle Veterinarmedizin 41(6): 851-6.  ISSN: 0003-9055.

            NAL Call Number:  41.8 Ex7

            Abstract:  Seven different hyperimmune serum samples from chickens or rabbits were conjugated with fluorescein isothiocyanate and reacted with reference influenza A strains. Conclusions are that direct immunofluorescence reliably detected avian influenza viruses and distinguished them from Newcastle disease virus. A diagnostic set of nine inactivated influenza A viruses is available, covering subtypes from H3 to H11.

            Descriptors:  fowl plague diagnosis, poultry diseases diagnosis, antibodies, viral analysis, influenza A virus avian immunology, poultry.

Wood, J.M., J. Mumford, G.C. Schild, R.G. Webster, and K.G. Nicholson (1986). Single-radial-immunodiffusion potency tests of inactivated influenza vaccines for use in man and animals. Developments in Biological Standardization 64: 169-77.  ISSN: 0301-5149.

            NAL Call Number:  QR180.3.D4

            Abstract:  Single-radial-immunodiffusion (SRD) provides a sensitive and reproducible in vitro assay for haemagglutinin (HA) concentration in inactivated influenza vaccines. The use of SRD for human influenza vaccine standardization and application for equine and avian influenza vaccines is discussed. In clinical trials, vaccine HA concentration measured by SRD has been shown to be directly related to antibody responses and to protection against challenge. The use of SRD may considerably reduce the usage of animals for potency testing of veterinary influenza vaccines.

            Descriptors:  influenza vaccine standards, vaccines, attenuated standards, antibodies, viral biosynthesis, chickens, horses, immunodiffusion standards, influenza prevention and control, influenza veterinary.

Woolcock, P.R., M.D. McFarland, S. Lai, and R.P. Chin (2001). Enhanced recovery of avian influenza virus isolates by a combination of chicken embryo inoculation methods. Avian Diseases 45(4): 1030-1035.  ISSN: 0005-2086.

            NAL Call Number:  41.8 Av5

            Descriptors:  avian influenza virus, diagnosis, embryos, ducks, pheasants, Psittaciformes, turkeys,  Galliformes, California, Illinois, Montana, Washington, United States, OECD countries, reviews.

Wunderwald, C. and R.K. Hoop (2002). Serological monitoring of 40 Swiss fancy breed poultry flocks. Avian Pathology 31(2):  157-62.  ISSN: 0307-9457.

            NAL Call Number:  SF995.A1A9

            Abstract:  Rapid serum agglutination, haemagglutination inhibition and enzyme-linked immunosorbent assays were used to screen Swiss fancy breed chicken flocks for antibodies against 12 avian infectious agents. For this purpose, 1,002 blood samples from 40 flocks were collected and tested. Ten percent of the samples were positive for Salmonella gallinarum-pullorum and 62.5% of the flocks were affected. More than 75% of the flocks had antibodies against Mycoplasma gallisepticum/Mycoplasma synoviae, infectious bronchitis, infectious bursal disease, avian encephalomyelitis, infectious chicken anaemia and reoviral arthritis. Low prevalence of antibodies was recorded for Salmonella enteritidis, avian influenza, avian leukosis and Newcastle disease (2.0 to 4.0%).

            Descriptors:  monitoring, immunologic veterinary, Mycoplasma infections veterinary, poultry, poultry diseases epidemiology, Salmonella infections, animal epidemiology, serologic tests veterinary, antibodies, bacterial blood, antibodies, viral blood, monitoring, immunologic methods, Mycoplasma infections epidemiology, poultry diseases immunology, prevalence, serologic tests methods, Switzerland epidemiology.

Xin ChaoAn (et al.)  (2000). Diagnosis, prevention and treatment of avian influenza. Poultry Husbandry and Diseases Control (7): 9-10.

            Descriptors:  diagnosis, prevention, treatment, avian influenza virus, fowl, Galliformes.

Yamaguchi, S., M. Mase, K. Tsukamoto, T. Imada, M. Narita, Y. Murakami, and Y. Yokoki (2000). Establishment of diagnostic methods for avian influenza. Bulletin of the National Institute of Animal Health (Japan) (106): 19-25.  ISSN: 0388-2403.

            NAL Call Number:  41.9 T572

            Descriptors:  chickens, avian influenza virus, diagnosis, PCR, birds, domestic animals, Galliformes, influenza virus, livestock, orthomyxoviridae, poultry, useful animals, viruses.

Zaides, V.M. and V.M. Zhdanov (1984). Reaktsiia tormozheniia gemaggliutinatsii kak indikator gumoral'nogo immuniteta pri grippe: o nekotorykh ogranicheniiakh metoda. Voprosy Virusologii 29(1): 4-8.  ISSN: 0507-4088.

            NAL Call Number:  448.8 P942

            Descriptors:  hemagglutination inhibition tests, influenza immunology, antibodies, viral analysis, antibody formation, antigens, viral analysis, chick embryo, complement fixation tests, hemagglutination, viral, influenza A virus avian immunology, influenza A virus immunology, neutralization tests.

Zhao JianMei, Wei Rong, Wang ZhiLiang, and Meng LiangYu (2003). Studies on application of one step multiple RT-PCR technique for detection of Newcastle disease, avian influenza and avian infectious bronchitis viruses. Chinese Journal of Animal Quarantine 20(1): 22-24.  ISSN: 1005-944X.

            Descriptors:  avian influenza virus, infectious bronchitis virus, Newcastle disease virus, polymerase chain reaction, China, Galliformes.

Zhou, E.M., M. Chan, R.A. Heckert, J. Riva, and M.F. Cantin (1998). Evaluation of a competitive ELISA for detection of antibodies against avian influenza virus nucleoprotein. Avian Diseases 42(3): 517-22.  ISSN: 0005-2086.

            NAL Call Number:  41.8 Av5

            Abstract:  A competitive enzyme-linked immunosorbent assay (C-ELISA) employing a baculovirus-expressed recombinant nucleoprotein and a monoclonal antibody was developed for the detection of antibodies to type A influenza virus nucleoprotein. The performance of the C-ELISA was evaluated by testing 756 chickens, 1123 turkeys, 707 emus, and 1261 ostriches, for a total of 3847 serum samples. Relative to the agar gel immunodiffusion (AGID) test, the C-ELISA had a sensitivity of 100% for all four species. The C-ELISA's sensitivity relative to the hemagglutination-inhibition (HI) test results was 100% for chicken, turkey, and emu and 96.2% for the ostrich serum samples. More than 90% of the AGID-negative/C-ELISA-positive serum samples were found positive by HI for at least one influenza serotype. The specificity of C-ELISA relative to AGID ranged from 85.5% to 99.8% for sera collected from these species. These results indicated that the C-ELISA was more sensitive and more specific than the AGID test and as sensitive and as specific as the HI test. The C-ELISA has the potential to replace the AGID test for screening sera from avian species, including ratites, for detection of antibodies to type A influenza virus.

            Descriptors:  antibodies, viral immunology, enzyme linked immunosorbent assay, influenza A virus avian immunology, nucleoproteins immunology, chickens immunology, chickens virology, emus immunology, emus virology, fowl plague immunology, fowl plague virology, ostriches immunology, ostriches virology, turkeys immunology, turkeys virology.

Zhu ChangGui  and Li YuMin  (1996). Diagnosis and management of an outbreak of avian influenza in exotic birds. Chinese Journal of Veterinary Medicine 22(1): 20.

            NAL Call Number:  SF604.C485

            Descriptors:  immunoprecipitation tests, serological surveys, avian influenza virus, diagnosis, management, outbreaks, pheasants, China.

Ziegler, T., H. Hall, A. Sanchez Fauquier, W.C. Gamble, and N.J. Cox (1995). Type- and subtype-specific detection of influenza viruses in clinical specimens by rapid culture assay. Journal of Clinical Microbiology 33(2): 318-21.  ISSN: 0095-1137.

            NAL Call Number:  QR46.J6

            Abstract:  A rapid culture assay which allows for the simultaneous typing and subtyping of currently circulating influenza A(H1N1), A(H3N2), and B viruses in clinical specimens was developed. Pools of monoclonal antibodies (MAbs) against influenza A and B viruses and MAbs HA1-71 and HA2-76, obtained by immunizing mice with the denatured hemagglutinin subfragments HA1 and HA2 of influenza virus A/Victoria/3/75, were used for immunoperoxidase staining of antigens in infected MDCK cells. MAb HA1-71 reacted exclusively with influenza A viruses of the H3 subtype, while MAb HA2-76 reacted with subtypes H1, H3, H4, H6, H8, H9, H10, H11, and H12, as determined with 78 human, 4 swine, and 10 avian influenza virus reference strains subtyped by the hemagglutination inhibition test. To determine if the technique can be used as a rapid diagnostic test, 263 known influenza virus-positive frozen nasal or throat swabs were inoculated into MDCK cells. After an overnight incubation, the cells were fixed and viral antigens were detected by immunoperoxidase staining. Influenza A viruses of the H1 and H3 subtypes were detected in 31 and 113 specimens, respectively. The subtypes of 10 influenza A virus-positive specimens could not be determined because they contained too little virus. Influenza B viruses were detected in 84 specimens, and 25 specimens were negative. We conclude that this assay is a rapid, convenient, non-labor-intensive, and relatively inexpensive test for detecting, typing, and subtyping influenza viruses in clinical specimens.

            Descriptors:  influenza virology, orthomyxoviridae classification, orthomyxoviridae isolation and purification, virus cultivation methods, antibodies, monoclonal, antibodies, viral, cell line, disease outbreaks, dogs, evaluation studies, false negative reactions, immunoenzyme techniques, influenza diagnosis, influenza epidemiology, influenza A virus avian classification, avian immunology, avian isolation and purification, human classification, human immunology, human isolation and purification, porcine classification, porcine immunology, porcine isolation and purification, influenza B virus classification, influenza B virus immunology, influenza B virus isolation and purification, mice, orthomyxoviridae immunology,  serotyping, time factors.


Return to:    Contents