title: Adjunctive mecA PCR for routine detection of methicillin susceptibility in clinical isolates of coagulase-negative staphylococci.
abstract: Concerns over the reliability of routine sensitivity testing in coagulase-negative staphylococci often lead to the use of potentially less-effective antibiotics as few laboratories have access to routine tests for the mecA resistance gene. Although previous studies have shown a reasonable correlation between oxacillin disc and automated sensitivity testing, changing epidemiology and methodology dictate periodic reappraisal of these methods. In the present study, we evaluated two real-time PCR assays against novel targets in the mecA gene as an adjunct to routine susceptibility testing using the Vitek II AST-P620 card. All samples were further examined for the presence of the mecC gene. Of 118 strains of coagulase-negative staphylococci tested, 81 were oxacillin resistant and 37 oxacillin susceptible by the Vitek II assay compared with 103 positive and 15 negative by mecA PCR. In-house PCR results correlated well with a previously published reference PCR, though little correlation was found between mecA PCR or Vitek II and PBP 2a latex agglutination. Incubation conditions may have affected the accuracy of the latter test. None of the strains tested were mecC PCR positive. The inclusion of dual-target PCRs in the testing algorithm was inexpensive and offered the safest strategy for determining beta-lactam susceptibility in coagulase-negative staphylococci in our laboratory.
authors.affiliation: Department of Medical Microbiology, King's College Hospital, London, United Kingdom.
authors.forename: Chandanjit Kaur
authors.initials: CK
authors.lastname: Nijjar
authors.forename: Melvyn Howard
authors.initials: MH
authors.lastname: Smith
authors.forename: Ian Joseph
authors.initials: IJ
authors.lastname: Eltringham
mesh_terms: D000900:Anti-Bacterial Agents / Q000494:pharmacology; D001426:Bacterial Proteins / Q000235:genetics; D003030:Coagulase / Q000235:genetics*; D005798:Genes, Bacterial / Q000235:genetics; D007841:Latex Fixation Tests / Q000379:methods; D008712:Methicillin / Q000494:pharmacology; D016106:Methicillin Resistance / Q000235:genetics*; D008826:Microbial Sensitivity Tests / Q000379:methods; D010068:Oxacillin / Q000494:pharmacology; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods; D015203:Reproducibility of Results; D013203:Staphylococcal Infections / Q000175:diagnosis* / Q000188:drug therapy / Q000382:microbiology*; D013210:Staphylococcus / Q000187:drug effects / Q000235:genetics* / Q000302:isolation & purification*; D047090:beta-Lactams / Q000494:pharmacology
chemical_list: D000900:Anti-Bacterial Agents; D001426:Bacterial Proteins; D003030:Coagulase; D047090:beta-Lactams; D008712:Methicillin; D010068:Oxacillin