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Journal of Clinical Microbiology, December 2004, p. 5731-5738, Vol. 42, No. 12
0095-1137/04/$08.00+0     DOI: 10.1128/JCM.42.12.5731-5738.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Phylogenetic Analysis and PCR-Restriction Fragment Length Polymorphism Identification of Campylobacter Species Based on Partial groEL Gene Sequences

R. I. Kärenlampi,* T. P. Tolvanen, and M.-L. Hänninen

Department of Food and Environmental Hygiene, Faculty of Veterinary Medicine, University of Helsinki, Helsinki, Finland

Received 5 July 2004/ Returned for modification 17 August 2004/ Accepted 30 August 2004

The phylogeny of 12 Campylobacter species and reference strains of Arcobacter butzleri and Helicobacter pylori was studied based on partial 593-bp groEL gene sequences. The topology of the phylogenetic neighbor-joining tree based on the groEL gene was similar to that of the tree based on the 16S rRNA gene. However, groEL was found to provide a better resolution for Campylobacter species, with lower interspecies sequence similarities (range, 65 to 94%) compared with those for the 16S rRNA gene (range, 90 to 99%) and high intraspecies sequence similarities (range, 95 to 100%; average, 99%). A new universal reverse primer that amplifies a 517-bp fragment of the groEL gene was developed and used for PCR-restriction fragment length polymorphism (PCR-RFLP) analysis of 68 strains representing 11 Campylobacter species as well as reference strains of A. butzlerii and H. pylori. Digestion with the AluI enzyme discriminated all Campylobacter species included in the study but showed more intraspecies diversity than digestion with the ApoI enzyme. A hippurate-negative variant of Campylobacter jejuni with a high level of groEL sequence similarity to both C. jejuni (96%) and C. coli (94%) gave a unique AluI profile and an ApoI profile identical to those of other C. jejuni strains. In conclusion, groEL gene sequencing and PCR-RFLP analysis are recommended as valuable tools for the identification of Campylobacter species.

* Corresponding author. Mailing address: Department of Food and Environmental Hygiene, Faculty of Veterinary Medicine, P.O. Box 66, University of Helsinki, FI-00014 Helsinki, Finland. Phone: 358-9-19157117. Fax: 358-9-19157101. E-mail: rauni.karenlampi{at}helsinki.fi.

Journal of Clinical Microbiology, December 2004, p. 5731-5738, Vol. 42, No. 12
0095-1137/04/$08.00+0     DOI: 10.1128/JCM.42.12.5731-5738.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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