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Journal of Clinical Microbiology, January 2003, p. 15-26, Vol. 41, No. 1
0095-1137/03/$08.00+0     DOI: 10.1128/JCM.41.1.15-26.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Comparative Genotyping of Campylobacter jejuni by Amplified Fragment Length Polymorphism, Multilocus Sequence Typing, and Short Repeat Sequencing: Strain Diversity, Host Range, and Recombination

Leo M. Schouls,1* Sanne Reulen,1 Birgitta Duim,2 Jaap A. Wagenaar,2 Rob J. L. Willems,1 Kate E. Dingle,3 Frances M. Colles,3 and Jan D. A. Van Embden1

Research Laboratory for Infectious Diseases, National Institute of Public Health and the Environment, Bilthoven,1 Division of Infectious Disease and Food Chain Quality, Institute for Animal Science and Health, Lelystad, The Netherlands,2 The Peter Medawar Building for Pathogen Research and Department of Zoology, Oxford University, Oxford OX1 3FY, England3

Received 9 July 2002/ Returned for modification 6 September 2002/ Accepted 18 October 2002

Three molecular typing methods were used to study the relationships among 184 Campylobacter strains isolated from humans, cattle, and chickens. All strains were genotyped by amplified fragment length polymorphism (AFLP) analysis, multilocus sequence typing (MLST), and sequence analysis of a genomic region with short tandem repeats designated clustered regularly interspaced short palindromic repeats (CRISPRs). MLST and AFLP analysis yielded more than 100 different profiles and patterns, respectively. These multiple-locus typing methods resulted in similar genetic clustering, indicating that both are useful in disclosing genetic relationships between Campylobacter jejuni isolates. Group separation analysis of the AFLP analysis and MLST data revealed an unexpected association between cattle and human strains, suggesting a common source of infection. Analysis of the polymorphic CRISPR region carrying short repeats allowed about two-thirds of the typeable strains to be distinguished, similar to AFLP analysis and MLST. The three methods proved to be equally powerful in identifying strains from outbreaks of human campylobacteriosis. Analysis of the MLST data showed that intra- and interspecies recombination occurs frequently and that the role of recombination in sequence variation is 50 times greater than that of mutation. Examination of strains cultured from cecum swabs revealed that individual chickens harbored multiple Campylobacter strain types and that some genotypes were found in more than one chicken. We conclude that typing of Campylobacter strains is useful for identification of outbreaks but is probably not useful for source tracing and global epidemiology because of carriage of strains of multiple types and an extremely high diversity of strains in animals.

* Corresponding author. Mailing address: Research Laboratory for Infectious Diseases, National Institute of Public Health and the Environment, P.O. Box 1, 3720 BA Bilthoven, The Netherlands. Phone: 31302742121. Fax: 31302744449. E-mail: LM.Schouls{at}rivm.nl.

Journal of Clinical Microbiology, January 2003, p. 15-26, Vol. 41, No. 1
0095-1137/03/$08.00+0     DOI: 10.1128/JCM.41.1.15-26.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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