The reporting of vancomycin resistance in Staphylococcus spp. has enormous therapeutic and epidemiological consequences. We evaluated Vitek software version 2.01 by the CLSI-recommended broth dilution method as well as the CDC-recommended vancomycin screen agar and Etest for vancomycin susceptibility testing of Staphylococcus spp. Of the 105 isolates of Staphylococci tested by the above methods, Vitek 2.01 gave 16 (15%) a false vancomycin intermediate/resistant phenotype. Laboratories using automated systems for routine microbiological susceptibility testing must confirm such resistance by validated methods.