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J Med Microbiol 57 (2008), 432-438; DOI: 10.1099/jmm.0.47628-0
© 2008 Society for General Microbiology
ISSN 1473-5644

PCR-RFLP assay for species and subspecies differentiation of the Streptococcus bovis group based on groESL sequences

Hsiao-Jan Chen1, Jui-Chang Tsai2,3, Tsung-Chain Chang4, Wei-Chun Hung1, Sung-Pin Tseng1, Po-Ren Hsueh5 and Lee-Jene Teng1,5

1 Department of Clinical Laboratory Sciences and Medical Biotechnology, National Taiwan University College of Medicine, Taipei, Taiwan

2 Center for Optoelectronic Biomedicine, National Taiwan University College of Medicine, Taipei, Taiwan

3 Division of Neurosurgery, Department of Surgery, National Taiwan University Hospital, Taipei, Taiwan

4 Department of Medical Laboratory Science and Biotechnology, School of Medicine, National Cheng Kung University, Tainan, Taiwan

5 Department of Laboratory Medicine, National Taiwan University Hospital, Taipei, Taiwan

Correspondence
Lee-Jene Teng
ljteng{at}ntu.edu.tw

Received 13 September 2007
Accepted 5 December 2007

The sequence diversity of groESL genes among Streptococcus bovis group isolates was analysed, including five reference strains and 36 clinical isolates. Phylogenetic analysis of the groES and groEL sequences showed that the isolates that belonged to the same species or subspecies usually clustered together. The intergenic spacer region between groES and groEL was variable in size (67–342 bp) and sequence and appeared to be a unique marker for species or subspecies determination. Sequence similarities of the groESL genes among species and subspecies ranged from 84.2 to 99.0 % in groES, and from 88.0 to 99.0 % in groEL. Based on the sequences determined, a Streptococcus bovis group-specific PCR assay was developed, which may provide an alternative means of distinguishing the bovis group from other viridans streptococci. Restriction digestion of the amplicon with AclI further differentiated the species and subspecies.

Abbreviations: ITS, intergenic spacer.

The GenBank/EMBL/DDBJ accession numbers for the groESL gene sequences reported in this study are given in Supplementary Table S1 in JMM Online.

Tables showing the GenBank/EMBL/DDBJ accession numbers for the groES and groEL gene sequences of strains used in this study and the nucleotide lengths and sequences of groES–groEL spacer regions are available as supplementary material with the online version of this paper.






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