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Data from four recent studies (S. H. Goh et al., J. Clin. Microbiol. 36:2164–2166, 1998; S. H. Goh et al., J. Clin. Microbiol. 34:818–823, 1996; S. H. Goh et al., J. Clin. Microbiol. 35:3116–3121, 1997; A. Y. C. Kwok et al., Int. J. Syst. Bacteriol. 49:1181–1192, 1999) suggest that an approximately 600-bp region of the chaperonin 60 (Cpn60) gene, amplified by PCR with a single pair of degenerate primers, has utility as a potentially universal target for bacterial identification (ID). This Cpn60 gene ID method correctly identified isolates representative of numerous staphylococcal species andStreptococcus iniae , a human and animal pathogen. We report herein that this method enabled us to distinguish clearly between 17Enterococcus species (Enterococcus asini ,Enterococcus rattus ,Enterococcus dispar ,Enterococcus gallinarum ,Enterococcus hirae ,Enterococcus durans ,Enterococcus cecorum ,Enterococcus faecalis ,Enterococcus mundtii ,Enterococcus casseliflavus ,Enterococcus faecium ,Enterococcus malodoratus ,Enterococcus raffinosus ,Enterococcus avium ,Enterococcus pseudoavium ,Enterococcus new sp. strain Facklam, andEnterococcus saccharolyticus ), andVagococcus fluvialis ,Lactococcus lactis , andLactococcus garvieae . From 123 blind-tested samples, only two discrepancies were observed between the Facklam and Collins phenotyping method (R. R. Facklam and M. D. Collins, J. Clin. Microbiol. 27:731–734, 1989) and the Cpn60 ID method. In each case, the discrepancies were resolved in favor of the Cpn60 ID method. The species distributions of the 123 blind-tested isolates wereEnterococcus new sp. strain Facklam (ATCC 700913), 3;E. asini , 1;E. rattus , 4;E. dispar , 2;E. gallinarum , 20;E. hirae , 9;E. durans , 9;E. faecalis , 12;E. mundtii , 3;E. casseliflavus , 8;E. faecium , 25;E. malodoratus , 3;E. raffinosus , 8;E. avium , 4;E. pseudoavium , 1; an unknownEnterococcus clinical isolate, sp. strain R871;Vagococcus fluvialis , 4;Lactococcus garvieae , 3;Lactococcus lactis , 3;Leuconostoc sp., 1; andPediococcus sp., 1. The Cpn60 gene ID method, coupled with reverse checkerboard hybridization, is an effective method for the identification ofEnterococcus and related organisms.

journal of clinical microbiology

Identification of Enterococcus Species and Phenotypically Similar Lactococcus and Vagococcus Species by Reverse Checkerboard Hybridization to Chaperonin 60 Gene Sequences