Diversity of methicillin-resistant Staphylococcus aureus isolated in a Canadian Hospital

Three neonates and three other patients located elsewhere in the hospital became infected with Staphylococcus aureus. Initial automated microdilution susceptibility testing with oxacillin and disk diffusion testing with amoxicillin-clavulanic acid indicated the isolates had borderline oxacillin resistance (MICs 4 micrograms/ml), presumably due to hyperproduction of beta-lactamase. Chromosomal DNA restriction fingerprinting and phage typing revealed the neonatal isolates to be identical; whereas, the other patients were infected with three different strains. Further analysis of the four strains by Southern hybridization with a mecA specific oligoprobe and a quantitative beta-lactamase assay demonstrated that two strains carried the mecA gene (coding for low affinity penicillin-binding protein 2a), and two strains were hyperproducers of beta-lactamase, including one which was mecA gene positive. One strain neither carried the mecA gene nor hyperproduced beta-lactamase. The two mecA gene positive strains displayed oxacillin MICs of 16 micrograms/ml on dilution susceptibility testing in 4% NaCl supplemented Mueller-Hinton agar. Hence, they were considered intrinsically methicillin-resistant Staphylococcus aureus. Both oxacillin and amoxicillin-clavulanic acid MICs were increased on NaCl supplementation. Results of amoxicillin-clavulanic acid disk diffusion susceptibility testing did not correlate with quantitative beta-lactamase production. It is recommended that clinical laboratories do not use amoxicillin-clavulanic acid disk diffusion assays to differentiate suspected borderline resistance due to beta-lactamase hyperproduction from mecA gene expression of PBP-2a since additional mechanisms may account for resistance.