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Journal of Clinical Microbiology, December 2002, p. 4612-4618, Vol. 40, No. 12
0095-1137/02/$04.00+0     DOI: 10.1128/JCM.40.12.4612-4618.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Strain Typing of Borrelia burgdorferi, Borrelia afzelii, and Borrelia garinii by Using Multiple-Locus Variable-Number Tandem Repeat Analysis

Jason Farlow,1 Danielle Postic,2 Kimothy L. Smith,1 Zack Jay,1 Guy Baranton,2 and Paul Keim1*

Department of Biological Sciences, Northern Arizona University, Flagstaff, Arizona 86011-5640,1 Institut Pasteur, Paris, France2

Received 12 February 2002/ Returned for modification 31 March 2002/ Accepted 2 September 2002

Human Lyme borreliosis (LB) is the most prevalent arthropod-borne infection in temperate climate zones around the world and is caused by Borrelia spirochetes. We have identified 10 variable-number tandem repeat (VNTR) loci present within the genome of Borrelia burgdorferi and subsequently developed a multiple-locus VNTR analysis (MLVA) typing system for this disease agent. We report here the successful application of MLVA for strain discrimination among a group of 41 globally diverse Borrelia isolates including B. burgdorferi, B. afzelii, and B. garinii. PCR assays displayed diversity at these loci, with total allele numbers ranging from two to nine and Nei's diversity (D) values ranging from 0.10 to 0.87. The average D value was 0.53 across all VNTR loci. A clear correlation exists between the repeat copy number and the D value (r = 0.62) or the number of alleles (r = 0.93) observed across diverse strains. Cluster analysis by the unweighted pair-group method with arithmetic means resolved the 30 observed unique Borrelia genotypes into five distinct groups. B. burgdorferi, B. afzelii, and B. garinii clustered into distinct affiliations, consistent with current 16S rRNA phylogeny studies. Genetic similarity and diversity suggest that B. afzelii and B. garinii are close relatives and were perhaps recently derived from B. burgdorferi. MLVA provides both phylogenetic relationships and additional resolution to discriminate among strains of Borrelia species. This new level of strain identification and discrimination will allow more detailed epidemiological and phylogenetic analysis in future studies.

* Corresponding author. Mailing address: Department of Biological Sciences, Northern Arizona University, P.O. Box 5640, Flagstaff, AZ 86011-5640. Phone: (928) 523-1078. Fax: (928) 523-0639. E-mail: Paul.Keim{at}nau.edu.

Journal of Clinical Microbiology, December 2002, p. 4612-4618, Vol. 40, No. 12
0095-1137/02/$04.00+0     DOI: 10.1128/JCM.40.12.4612-4618.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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