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1Departments of Medicine and Microbiology and Immunology, University of British Columbia, Vancouver, BC, Canada
2Departments of Medicine, University of British Columbia, Vancouver, BC, Canada
3Vancouver Hospital Health Sciences Centre, 2733 Heather Street, Vancouver, BC, Canada
4Department of Chemistry, University of Alberta, Edmonton, AB, Canada
5 Division of Infectious Diseases, G. F. Strong Research Laboratories and the Canadian Bacterial Diseases Network, Vancouver, BC, Canada
Author for correspondence: Anthony W. Chow. Tel: + 1 604 875 4148. Fax: + 1 604 875 4013. e-mail: tonychow{at}interchange.ubc.ca
ABSTRACT
The phylogenetic relationships among 36 validly described species or subspecies within the genus Staphylococcus were investigated by cloning an sequencing their 60 kDa heat-shock protein (HSP60) genes using a set of universal degenerate HSP60 PCR primers. The cloned partial HSP60 DNA sequences from nine Staphylococcus aureus strains were highly conserved (97-100% DNA sequence similarity; mean 98%), indicating that the HSP60 gene of multiple isolates within the same species have little microheterogeneity. At the subspecies level, DNA sequence similarity among members of S. aureus, Staphylococcus schleiferi, Staphylococcus cohnii and Staphylococcus capitis ranged from 91 to 98%. At the interspecies level, sequence similarity among 23 distinct species of staphylococci ranged from 7 to 93% (mean 82%). By comparison, the highest sequence similarity of Bacill subtilis and Escherichia coli with members within the genus Staphylococcus was only 70 and 59%, respectively. Importantly, phylogenetic analysis based on the neighbour-joining distance method revealed remarkable concordance between the tree derived from partial HSP60 gene sequences and that based on genomic DNA-DNA hybridization, while 16S rRNA gene sequences correlated less well. The results demonstrate that DNA sequences from the highly conserved and ubiquitous HSP60 gene offer a convenient and accure tool for species-specific identification and phylogenetic analysis of staphylococci.
Key Words: Staphylococcus • HSP60 gene sequences • 16S rRNA gene sequences • DNA-DNA hybridization • phylogenetic analysis
The GenBank accession numbers for the sequences reported in this paper are AF033622, AF036322-AF036324, AF053568-AF053587, AF060184-AF060191 and U92809.
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