Background and Objective: Vancomycin is frequently the antibiotic of choice for the treatment of infections caused by methicillin-resistant Staphylococcus aureus (MRSA). For the last years, the incidence of vancomycin-intermediate S. aureus (VISA) and vancomycin-resistant S. aureus (VRSA) has been increased in various parts of the world. The present study was carried out to determine the presence of VISA and VRSA in Tehran. Materials and Methods: A total of 164 S. aureus strains were isolated from clinical specimens in four university-affiliated hospitals in Tehran from November 2006 to June 2007. Minimum inhibitory concentration (MIC) of vancomycin of isolates was determined by agar dilution method. Vancomycin (6 mg/l) screen agar plate method and E-test were used to confirm presence of resistance to vancomycin. Disc diffusion agar test was also used to detect resistance to other antimicrobial agents. Results: Only one VRSA(MIC 256 mg/l) was detected and three strains with MIC 4 mg/l considered VISA according to recent CLSI breakpoints for vancomycin. Only VRSA strain had shown growth on vancomycin screen agar plate and was also resistant to several antimicrobial agents but susceptible to quinupristin/dalfopristin, linezolid, chloramphenicol, mupirocin and cotrimoxazole. Isolated VISA were also multi-resistant but showed susceptibility to quinupristin/dalfopristin, linezolid, chloramphenicol and mupirocin. Conclusion: Detection of vancomycin resistance in Iranian S. aureus isolates emphasizes the challenges confronted by the infection control specialists in hospitals in Iran as well as causing problems in the treatment of patients with S. aureus infections.