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Journal of Clinical Microbiology, November 2000, p. 3953-3959, Vol. 38, No. 11
0095-1137/00/$04.00+0

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

Swee Han Goh,1,2,* Richard R. Facklam,3 Michelle Chang,1 Janet E. Hill,5 Gregory J. Tyrrell,4 Emma C. M. Burns,2 David Chan,2 Cheng He,2 Tazim Rahim,2 Carol Shaw,2 and Sean M. Hemmingsen5

Department of Pathology & Laboratory Medicine, The University of British Columbia,1 and Laboratory Services, British Columbia Centre for Disease Control Society,2 Vancouver, British Columbia, National Centre for Streptococcus, Edmonton, Alberta,4 and National Research Council Canada, Plant Biotechnology Institute, Saskatoon, Saskatchewan,5 Canada, and Centers for Disease Control and Prevention, Atlanta, Georgia3

Received 26 May 2000/Returned for modification 27 July 2000/Accepted 28 August 2000

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 and Streptococcus iniae, a human and animal pathogen. We report herein that this method enabled us to distinguish clearly between 17 Enterococcus 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, and Enterococcus saccharolyticus), and Vagococcus fluvialis, Lactococcus lactis, and Lactococcus 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 were Enterococcus 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 unknown Enterococcus clinical isolate, sp. strain R871; Vagococcus fluvialis, 4; Lactococcus garvieae, 3; Lactococcus lactis, 3; Leuconostoc sp., 1; and Pediococcus sp., 1. The Cpn60 gene ID method, coupled with reverse checkerboard hybridization, is an effective method for the identification of Enterococcus and related organisms.

* Corresponding author. Mailing address: Dept. of Pathology & Laboratory Medicine, The University of British Columbia, and Laboratory Services, British Columbia Centre for Disease Control Society, 655W 12th Ave., Vancouver, British Columbia V5Z 4R4, Canada. Phone: (604) 660-6005. Fax: (604) 660-0403. E-mail: shgoh{at}interchange.ubc.ca.

Journal of Clinical Microbiology, November 2000, p. 3953-3959, Vol. 38, No. 11
0095-1137/00/$04.00+0


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