Performance of different methods of oxacillin resistance detection in atypic strains of Staphylococcus aureus

Seventy-three of aminoglycoside-susceptible methicillin-resistant Staphylococcus aureus (AS-MRSA) and 12 kanamycin-tobramycin-resistant methicillin-susceptible S. aureus (KTR-MSSA) isolates were phenotypically and genotypically examined for methicillin susceptibility. The AS-MRSA profile represents 8.3% of MRSA strains and the KTR-MSSA profile represents 1.38% of MSSA strains. The diffusion method using the 5 microg oxacillin and 30 microg cefoxitin discs on Mueller-Hinton Agar (MHA) with and without NaCl, the incubation at 35 degrees C or 30 degrees C for 24 or 48 hours respectively, and the determining oxacillin MICs by E-test (AES, Combourg, France) were performed and used as phenotypic methods. We also used the mecA gene PCR which was considered as the "gold standard" for methicillin resistance detection, and the Slidex MRSA Detection (bioMérieux) that detect the presence of mecA gene product (PBP 2a). To increase the level of PBP 2a expression, the 30 microg cefoxitin disc was used as an inducer. All the AS-MRSA strains (100%) were detected by the cefoxitin disc in all conditions and by the oxacillin disc on MHA with 2% of NaCl at 35 degrees C. Without NaCl, the sensitivity fell to 97,2% by oxacillin disc. The oxacillin MICs for these isolates ranged from 2 to 128 mg/l. The mecA gene determinant and its product PBP 2a were detected in all AS-MRSA strains. All KTR-MSSA strains were phenotypically methicillin-susceptible and oxacillin MICs were below or borderline of breakpoint (< or =2 mg/l). The mecA gene determinant and its product were detected in one strain which was considered to be the most heterogeneous of those tested.