VANNUFFLE P, LOCKMAN L, REMACLE J, GALA JL; Interscience Conference on Antimicrobial Agents and Chemotherapy.
Abstr Intersci Conf Antimicrob Agents Chemother Intersci Conf Antimicrob Agents Chemother. 1999 Sep 26-29; 39: 208 (abstract no. 875).
Catholic Univ. of Louvain, Brussels, BELGIUM
A sandwich enzyme-linked oligo sorbent assay (ELOSA) was set up to conjointly discriminate between the most clinically relevant staphylococal species : Staphylococcus aureus, St. epidermidis, St. haemolyticus, St. hominis and St. Saprophyticus, together with the methicillin resistance genotype. Species identification was based on interspecies nucleotides sequences variations of the staphylococcal femA gene, a key component of the peptidoglycan biosynthesis. Methicillin resistance was assessed by detecting the highly conserved mecA determinant. femA species-specific and mecA specific capture probes were produced from the respective genes cloned into TOPO XL PCR (Invitrogen). Each capture probe was immobilized onto a distinct microplate well. Targeted amplicons were produced from reference strains or clinical samples of interest by the use of a pair of universal primers matching conserved portions of the femA and mecA. Selective hybridization of the targets was restricted to corresponding species-specific or resistance-specific capture probe. Captured targets were colorimetrically revealed by a biotinylated synthetic detection probe and a streptavidin-peroxidase conjugate. Despite high homology (up to 80%) between the five femA sequences, ELOSA was specific on the 5 ATCC reference strains as well as on a panel of methicillin-susceptible and -resistant staphylococcal clinical isolates representing all five species (n= 75). Sensitivity was confirmed by limiting dilutions of DNA from reference strains and clinical samples of interest.
Publication Types:
Keywords:
- Aluminum Hydroxide
- Base Sequence
- Carbonates
- Case-Control Studies
- Coagulase
- DNA Primers
- Hexosyltransferases
- Link
- Methicillin Resistance
- Muramoylpentapeptide Carboxypeptidase
- Oligonucleotides
- Peptidyl Transferases
- Polymerase Chain Reaction
- Sensitivity and Specificity
- Staphylococcus
- Staphylococcus aureus
- enzymology
- genetics
Other ID:
UI: 102245146
From Meeting Abstracts