Detection and expression of methicillin/oxacillin resistance in multidrug-resistant and non-multidrug-resistant Staphylococcus aureus in Central Sydney, Australia

J Antimicrob Chemother. 2002 May;49(5):793-801. doi: 10.1093/jac/dkf021.

Abstract

Ninety clinical Staphylococcus aureus isolates from separate patients were examined phenotypically and genotypically for susceptibility to methicillin/oxacillin. Thirty were methicillin/oxacillin susceptible and 60 were methicillin and oxacillin resistant (MRSA). The 60 MRSA isolates examined were subdivided into two groups according to their antibiotic profiles and comprised 30 non-multidrug-resistant (NMDR) isolates, resistant to less than two non-beta-lactam antibiotics, and 30 multidrug-resistant (MDR) isolates, resistant to three or more non-beta-lactam antibiotics. Phenotypic and genotypic analysis of methicillin/oxacillin showed that despite use of the guidelines published by the NCCLS for the testing of S. aureus susceptibility to methicillin/oxacillin, MIC values of some NMDR MRSA isolates fell below the NCCLS-recommended breakpoints. Etest strips failed to detect two NMDR MRSA isolates tested with oxacillin and four tested with methicillin. Lowering the NCCLS-recommended oxacillin screen agar concentration from 6 to 2 mg/L and temperature of incubation to 30 degrees C, improved the specificity and sensitivity of NMDR MRSA detection from 87% to 100%. On PFGE analysis these NMDR MRSA strains were genotypically different. Genotypic tests, such as multiplex PCR for the mecA/nuc genes and DNA hybridization for the mecA gene, or phenotypic monoclonal antibody-based tests to detect penicillin-binding protein 2a (PBP2a) offer advantages for problematic isolates in detecting or confirming low-level phenotypic heterogeneous mecA expression of oxacillin and methicillin resistance in NMDR MRSA.

MeSH terms

  • Bacterial Proteins*
  • DNA, Bacterial / genetics
  • DNA, Bacterial / isolation & purification
  • Drug Resistance, Multiple
  • Endonucleases / genetics
  • Genotype
  • Methicillin Resistance / physiology*
  • Microbial Sensitivity Tests
  • Micrococcal Nuclease*
  • New South Wales
  • Oxacillin / pharmacology*
  • Penicillin Resistance / physiology*
  • Penicillin-Binding Proteins
  • Penicillins / pharmacology*
  • Phenotype
  • Reverse Transcriptase Polymerase Chain Reaction
  • Staphylococcus aureus / drug effects*

Substances

  • Bacterial Proteins
  • DNA, Bacterial
  • Penicillin-Binding Proteins
  • Penicillins
  • mecA protein, Staphylococcus aureus
  • nuc protein, staphylococcus
  • Endonucleases
  • Micrococcal Nuclease
  • Oxacillin