Treatment failures with vancomycin prompted us to investigate the phenomenon of tolerance to glycopeptides in recent clinical isolates of Staphylococcus aureus. We used both MBC/MIC determinations and time-kill measurements to study tolerance to vancomycin and teicoplanin in 35 blood or heart valve isolates of S. aureus from patients with endocarditis or bacteraemia. There was generally good agreement between vancomycin tolerance indicated by an MBC:MIC ratio of > or =32 and by < or =90% kill after 6 h incubation in the presence of 20 mg/L vancomycin. However, two isolates were tolerant according to their MBC:MIC ratios but non-tolerant as judged by time-kill measurements. Seven of 15 methicillin-resistant S. aureus (MRSA) isolates but only two of 20 methicillin-susceptible ones were tolerant as judged by time-kill experiments (chi2 = 4.27 with Yates' correction, P = 0.04). Seven of the 16 isolates from patients with endocarditis were tolerant, compared with only two of the 19 isolates from patients with other conditions (chi2 = 3.43 with Yates' correction, P = 0.06). Within the endocarditis and non-endocarditis subgroups, tolerance was associated more frequently with methicillin resistance than with susceptibility, but the numbers were too small for the differences to be statistically significant. Most of the vancomycin-tolerant isolates were also tolerant to teicoplanin. We conclude that glycopeptide tolerance is a real phenomenon in S. aureus, particularly amongst MRSA isolates, and can be reliably determined by our method of time-kill analysis. Tolerance may compromise glycopeptide therapy of serious S. aureus infection and should be taken into account when deciding treatment.