A study was undertaken to develop an easy-to-use and cost-effective algorithm for the detection and identification of vancomycin-resistant enterococci (VRE) in surveillance cultures, because the incidence of VRE outbreaks in institutions across Canada has made continuous surveillance a necessity. Enterococcus faecium and Enterococcus faecalis carry transferable resistance genes and hence are a problem for infection control. In laboratory surveillance, however, Enterococcus gallinarum and Enterococcus casseliflavus, which exhibit low-level nontransferable resistance, are also encountered and often create confusion in identification. Included in this study were a total of 218 strains of enterococci and other streptococci isolated from surveillance cultures. Conventional methods were used to determine their biochemical activities, and speciation was attempted in 121 strains using a rapid multiplex polymerase chain reaction (PCR) method that utilized primers for the vanA, B, C1, C2/C3 and ddl genes. The results indicated that by using only a few tests (Gram stain, pyrrolidonyl arylamidase activity, motility, xylose and methyl-alpha-D-glucopyranoside utilization), Enterococcus faecium/faecalis strains could be accurately differentiated from Enterococcus gallinarum/casseliflavus strains. For the 121 strains on which PCR was performed, there was a 100% correlation with the biochemical identification, with the added advantage that the presence of van genes could be determined at the same time. The cost of identification using minimal biochemical testing and PCR was less than that of identification using automated systems or a battery of conventional biochemical methods. The algorithm presented here may be used in the microbiology laboratory.