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Journal of Clinical Microbiology, September 2001, p. 3186-3192, Vol. 39, No. 9
0095-1137/01/$04.00+0   DOI: 10.1128/JCM.39.9.3186-3192.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Francisella tularensis Strain Typing Using Multiple-Locus, Variable-Number Tandem Repeat Analysis

Jason Farlow,1 Kimothy L. Smith,1 Jane Wong,2 Michelle Abrams,1 Michael Lytle,3 and Paul Keim1,*

Department of Biological Sciences, Northern Arizona University, Flagstaff, Arizona 86011-56401; Microbial Diseases Laboratory, California Department of Health Services, Berkeley, California 947042; and Oklahoma State Department of Health, Oklahoma City, Oklahoma 731173

Received 23 March 2001/Returned for modification 2 June 2001/Accepted 1 July 2001

Francisella tularensis, the etiological agent of tularemia, is found throughout the Northern hemisphere. After analyzing the F. tularensis genomic sequence for potential variable-number tandem repeats (VNTRs), we developed a multilocus VNTR analysis (MLVA) typing system for this pathogen. Variation was detected at six VNTR loci in a set of 56 isolates from California, Oklahoma, Arizona, and Oregon and the F. tularensis live vaccine strain. PCR assays revealed diversity at these loci with total allele numbers ranging from 2 to 20, and Nei's diversity index values ranging from 0.36 to 0.93. Cluster analysis identified two genetically distinct groups consistent with the current biovar classification system of F. tularensis. These findings suggest that these VNTR markers are useful for identifying F. tularensis isolates at this taxonomic level. In this study, biovar B isolates were less diverse than those in biovar A, possibly reflecting the history of tularemia in North America. Seven isolates from a recent epizootic in Maricopa County, Ariz., were identical at all VNTR marker loci. Their identity, even at a hypervariable VNTR locus, indicates a common source of infection. This demonstrates the applicability of MLVA for rapid characterization and identification of outbreak isolates. Future construction of reference databases will allow faster outbreak tracking as well as providing a foundation for deciphering global genetic relationships.

* Corresponding author. Mailing address: Department of Biological Sciences, Northern Arizona University, Flagstaff, AZ 86011-5640. Phone: (520) 523-1078. Fax: (520) 523-0639. E-mail: Paul.Keim{at}nau.edu.

Journal of Clinical Microbiology, September 2001, p. 3186-3192, Vol. 39, No. 9
0095-1137/01/$04.00+0   DOI: 10.1128/JCM.39.9.3186-3192.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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