BACKGROUND: Enterococci are important cause of nosocomial infections. Recently, vancomycin-resistant enterococci (VRE) has been increasingly reported as significant nosocomial pathogens. Therefore, accurate identification of enterococcal species is a prerequisite step for the appropriate antibiotic treatment and epidemiologic surveillance. We wanted to know the usefulness of PCR method compared with Vitek automatic identification system. METHODS: Totally 105 isolates were identified on the species level by Vitek (GPI card and software version R06.1), methyl-alpha-D-glucopyranoside test, and PCR methods. RESULTS: Among 105 enterococcal isolates, 59 were identified as E. faecium, 11 E. faecalis, 6 E. gallinarum by Vitek. But 29 isolates (28%) were unidentified. Subsequently all of these isolates were analyzed by PCR, the results of which were as follows: 17 E. faecium, 5 E. casseliflavus, 7 E. gallinarum. Two isolates identified as E. gallinarum by Vitek were reidentified as E. casseliflavus by PCR and other methods for phenotypic characterization. CONCLUSOIN: PCR method was more accurate and sensitive than Vitek for the identification of enterococci species.