Investigation of Reduced Susceptibility to Glycopeptides among Methicillin-Resistant Staphylococcus aureusIsolates from Patients in Ireland and Evaluation of Agar Screening Methods for Detection of Heterogeneously Glycopeptide-Intermediate S. aureus

ABSTRACTMethicillin-resistant Staphylococcus aureus(MRSA) isolates (n= 3,189) from 2,990 patients were investigated by agar screening and by the Etest macromethod for reduced susceptibility to glycopeptide. No vancomycin-resistant S. aureusor glycopeptide-intermediate S. aureus(GISA) isolates were detected, but 178 isolates were confirmed as hetero-GISA (hGISA) by vancomycin population analysis profile (vPAP)-area under the curve (AUC) ratio determination and/or teicoplanin PAP (tPAP) methods. Of 139 isolates detected using the recommended Etest macromethod cutoff values of =8 mg/liter for both vancomycin and teicoplanin or =12 mg/liter for teicoplanin alone, 73 were confirmed as hGISA by vPAP-AUC, 95 were confirmed as hGISA by tPAP, and 108 were confirmed as hGISA by both methods. An Etest macromethod cutoff value of 8 mg/liter for teicoplanin alone detected a further 70 hGISA (17 were confirmed by vPAP-AUC and 70 were confirmed by tPAP). Agar screening utilizing brain heart infusion (BHI) agar containing 6 mg of vancomycin/liter (BHIV6) and Mueller-Hinton (MH) agar containing 8 mg of teicoplanin/liter (MHT8) failed to detect hGISA. MH agar containing 5 mg of teicoplanin/liter (MHT5) and BHI containing 5 mg of teicoplanin/liter (BHIT5) were evaluated using 10-µl volumes of three inoculum concentrations (with densities equivalent to 0.5 and 2.0 McFarland turbidity standards and stationary-phase BHI broth subcultures [MHT50.5, MHT52.0, MHT5S, BHIT50.5, BHIT52.0, and BHIT5S]). The sensitivity of all methods except MHT50.5and MHT52.0was 100%. The specificity ranged from 4 to 82%. BHIT50.5yielded the best performance, with a specificity of 84% for detecting isolates with teicoplanin Etest macromethod values of =8 mg/liter. Screening on BHIT50.5is useful where screen-positive isolates are investigated with the Etest macromethod and confirmed by vPAP-AUC and tPAP. The prevalence of hGISA among patients with blood culture isolates recovered in Irish hospitals between 1999 and 2003 was 2.6%, whereas the prevalence among patients with isolates from all specimen sites collected during a 2-week survey in 1999 was 12%. The prevalence in one hospital decreased from 5.3% in 2003 to 1.5% in 2004.