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Journal of Clinical Microbiology, September 2001, p. 3140-3146, Vol. 39, No. 9
0095-1137/01/$04.00+0   DOI: 10.1128/JCM.39.9.3140-3146.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Extensive Allelic Variation among Francisella tularensis Strains in a Short-Sequence Tandem Repeat Region

Anders Johansson,1,2,3 Ingela Göransson,3 Pär Larsson,2,3 and Anders Sjöstedt2,3,*

Department of Clinical Microbiology, Infectious Diseases,1 and Department of Clinical Microbiology, Clinical Bacteriology,2 Umeå University, Umeå, and NBC Analysis, Swedish Defence Research Agency,3 Umeå, Sweden

Received 14 March 2001/Returned for modification 7 June 2001/Accepted 17 June 2001

Members of the genus Francisella and the species F. tularensis appear to be genetically very similar despite pronounced differences in virulence and geographic localization, and currently used typing methods do not allow discrimination of individual strains. Here we show that a number of short-sequence tandem repeat (SSTR) loci are present in F. tularensis genomes and that two of these loci, SSTR9 and SSTR16, are together highly discriminatory. Labeled PCR amplification products from the loci were identified by an automated DNA sequencer for size determination, and each allelic variant was sequenced. Simpson's index of diversity was 0.97 based on an analysis of 39 nonrelated F. tularensis isolates. The locus showing the highest discrimination, SSTR9, gave an index of diversity of 0.95. Thirty-two strains isolated from humans during five outbreaks of tularemia showed much less variation. For example, 11 of 12 strains isolated in the Ljusdal area, Sweden in 1995 and 1998 had identical allelic variants. Phenotypic variants of strains and extensively cultured replicates within strains did not differ, and, for example, the same allelic combination was present in 55 isolates of the live-vaccine strain of F. tularensis and another one was present in all 13 isolates of a strain passaged in animals. The analysis of short-sequence repeats of F. tularensis strains appears to be a powerful tool for discrimination of individual strains and may be useful for a detailed analysis of the epidemiology of this potent pathogen.

* Corresponding author. Mailing address: Department of Clinical Microbiology, Clinical Bacteriology, Umeå University, SE-901 85 Umeå, Sweden. Phone: 46 90 7851120. Fax: 46 90 7852225. E-mail: anders.sjostedt{at}climi.umu.se.

Journal of Clinical Microbiology, September 2001, p. 3140-3146, Vol. 39, No. 9
0095-1137/01/$04.00+0   DOI: 10.1128/JCM.39.9.3140-3146.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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