The performance of the Vitek System and the API 20S for identifying enterococci to the species level was evaluated in comparison with a conventional reference test scheme. A total of 369 isolates originally derived from a sample from 97 medical centers were tested, and the percentage of correctly identified organisms was (Vitek/API): 97.6%/79.2%, 95.3%/91.2%, and 86.0%/20.7% for Enterococcus faecalis, Enterococcus faecium, and Enterococcus non-faecalis-non-faecium species, respectively. In addition, the predictive values for species identification of the results reported by the systems were 98.1% for Vitek and 85.9% for API. The Vitek System GPI card (software version > or = 8.1) was considered to be very acceptable for enterococci identification, with errors principally among the rare Enterococcus raffinosus isolates.