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Journal of Clinical Microbiology, September 2001, p. 3326-3331, Vol. 39, No. 9
0095-1137/01/$04.00+0   DOI: 10.1128/JCM.39.9.3326-3331.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Determination of Enterococcus faecalis groESL Full-Length Sequence and Application for Species Identification

Lee-Jene Teng,1,2,* Po-Ren Hsueh,2 Yi-Hui Wang,1 Hsiao-Mann Lin,1 Kwen-Tay Luh,2 and Shen-Wu Ho1,2

School of Medical Technology, National Taiwan University College of Medicine,1 and Department of Laboratory Medicine, National Taiwan University Hospital,2 Taipei, Taiwan

Received 20 March 2001/Returned for modification 14 May 2001/Accepted 3 July 2001

Amplification of the partial Cpn60 (or GroEL) gene segment has been used for identification of many bacteria, including Enterococcus species. To obtain more sequence data from groESL genes of Enterococcus faecalis, the full-length sequence of the E. faecalis groESL genes containing groES (285 bp), spacer (57 bp), and groEL (1,626 bp) was determined. A database search of GenBank revealed that the deduced E. faecalis GroES and GroEL proteins show significant homology to the GroES and GroEL proteins of other bacteria. The GroEL (groEL) of E. faecalis had the highest identity with Streptococcus pneumoniae (81.8% amino acid sequence identity and 73.0% nucleotide sequence identity), followed by Lactococcus zeae, while GroES (groES) had 60.2% (64.6%) identity with Lactobacillus zeae and 58.5% (66.2%) identity with Lactococcus lactis, followed by 57.0% (65.5%) identity with Bacillus subtilis. Based on the groES sequence, an E. faecalis-specific PCR assay was developed, and this PCR assay was positive for all the E. faecalis strains tested. Dot blot hybridization using either groES or groEL as the probe distinguished E. faecalis clearly from other species, indicating that both genes can be used as suitable targets for E. faecalis identification. Moreover, broad-range PCR-restriction fragment length polymorphism of groESL was designed to differentiate eight commonly encountered Enterococcus species. The Enterococcus species of reference strains could be easily differentiated on the basis of restriction patterns produced by HaeIII and RsaI. The DNA-based assays developed in this study provide an alternative to currently used methods of identification for clinically important enterococcal species.

* Corresponding author. Mailing address: School of Medical Technology, National Taiwan University College of Medicine, No 1, Chang-Te St., Taipei, Taiwan. Phone: 886-2-23123456, ext. 6918. Fax: 886-2-23711574. E-mail: ljteng{at}ha.mc.ntu.edu.tw.

Journal of Clinical Microbiology, September 2001, p. 3326-3331, Vol. 39, No. 9
0095-1137/01/$04.00+0   DOI: 10.1128/JCM.39.9.3326-3331.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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