A total of 250 consecutive Staphylococcus aureus clinical isolates were collected during the period 1999-2000 from the five major hospitals of the district of Thessaly (Central Greece). Thirty seven (14.8%) of the isolates were mecA-positive (MRSA) in a PCR-based assay; all exhibited resistance to oxacillin (agar dilution MICs > or =4 mg/L) and were also resistant to multiple antibiotics. Most of the MRSA isolates had been collected in the intensive care units and the surgical wards of the participating hospitals in a sporadic fashion. The MRSA incidence found here was significantly lower than reported in previous studies from Greece. Molecular typing by PFGE showed that the MRSA isolates were distributed between three pulsotypes. Evaluation of various conventional methods for assessing methicillin resistance showed that oxacillin agar dilution and immunological detection of PBP2a with the Slidex MRSA Detection kit were the most reliable in this setting. Misclassifications of isolates exhibiting low-level resistance (oxacillin MIC 2-4 mg/L) occurred with the salt agar screen, the oxacillin disk diffusion and the ATB Staph System methods.