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J Med Microbiol 55 (2006), 393-399; DOI: 10.1099/jmm.0.46282-0
© 2006 Society for General Microbiology
ISSN 0022-2615

Identification of Campylobacter spp. and discrimination from Helicobacter and Arcobacter spp. by direct sequencing of PCR-amplified cpn60 sequences and comparison to cpnDB, a chaperonin reference sequence database

Janet E. Hill1,{dagger}, Ana Paccagnella2, Kee Law2, Pasquale L. Melito3, David L. Woodward3, Lawrence Price3, Amy H. Leung4, Lai-King Ng3, Sean M. Hemmingsen1 and Swee Han Goh2,4,5

1 National Research Council Plant Biotechnology Institute, Saskatoon, Saskatchewan, Canada

2 British Columbia Centre for Disease Control, 655W 12th Ave, Vancouver, BC, V5Z 4R4, Canada

3 Bacteriology and Enteric Diseases Program, National Microbiology Laboratory, Winnipeg, Manitoba, Canada

4 ,5 Dept of Pathology and Laboratory Medicine4 , and UBC CDC5 , University of British Columbia, BC, Canada

Correspondence
Swee Han Goh
shgoh{at}interchange.ubc.ca

Received 4 August 2005
Accepted 6 December 2005

A robust method for the identification of Campylobacter spp. based on direct sequencing of PCR-amplified partial cpn60 sequences and comparison of these to a reference database of cpn60 sequences is reported. A total of 53 Campylobacter isolates, representing 15 species, were identified and distinguished from phenotypically similar Helicobacter and Arcobacter strains. Pairwise cpn60 sequence identities between Campylobacter spp. ranged from 71 to 92 %, with most between 71 and 79 %, making discrimination of these species obvious. The method described overcomes limitations of existing PCR-based methods, which require time-consuming and complex post-amplification steps such as the cloning of amplification products. The results of this study demonstrate the potential for use of the reference chaperonin sequence database, cpnDB, as a tool for identification of bacterial isolates based on cpn60 sequences amplified with universal primers.

Abbreviations: BCCDC, British Columbia Centre for Disease Control; NML, National Microbiology Laboratory.

{dagger}Present address: Dept of Veterinary Microbiology, University of Saskatchewan, Saskatoon, SK, Canada.

The GenBank/EMBL/DDBJ accession numbers for the cpn60 sequences determined in this study are DQ059425–DQ059481.




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