This Article Full Text Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Reprints and Permissions Copyright Information Books from ASM Press MicrobeWorld Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Johansson, A. Articles by Keim, P. Search for Related Content PubMed PubMed Citation Articles by Johansson, A. Articles by Keim, P.

 Previous Article  |  Next Article 

Journal of Bacteriology, September 2004, p. 5808-5818, Vol. 186, No. 17
0021-9193/04/$08.00+0     DOI: 10.1128/JB.186.17.5808-5818.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Worldwide Genetic Relationships among Francisella tularensis Isolates Determined by Multiple-Locus Variable-Number Tandem Repeat Analysis

Anders Johansson,^1,2, Jason Farlow,^3, Pär Larsson,² Meghan Dukerich,³ Elias Chambers,³ Mona Byström,² James Fox,³ May Chu,⁴ Mats Forsman,² Anders Sjöstedt,⁵ and Paul Keim^3*

Divisions of Infectious Diseases,¹ Clinical Bacteriology, Department of Clinical Microbiology, Umeå University,⁵ Department of NBC Analysis, Swedish Defence Research Agency, Umeå, Sweden,² Department of Biological Sciences, Northern Arizona University, Flagstaff, Arizona,³ Centers for Disease Control and Prevention, Fort Collins, Colorado⁴

Received 29 January 2004/ Accepted 6 June 2004

The intracellular bacterium Francisella tularensis is the causative agent of tularemia and poses a serious threat as an agent of bioterrorism. We have developed a highly effective molecular subtyping system from 25 variable-number tandem repeat (VNTR) loci. In our study, multiple-locus VNTR analysis (MLVA) was used to analyze genetic relationships and potential population structure within a global collection of 192 F. tularensis isolates, including representatives from each of the four subspecies. The VNTR loci displayed between 2 and 31 alleles with Nei's diversity values between 0.05 and 0.95. Neighbor-joining cluster analysis of VNTR data revealed 120 genotypes among the 192 F. tularensis isolates, including accurate subspecies identification. F. tularensis subsp. tularensis (type A) isolates showed great diversity at VNTR loci, while F. tularensis subsp. holarctica (type B) isolates showed much lower levels despite a much broader geographical prevalence. The resolution of two distinct clades within F. tularensis subsp. tularensis (designated A.I and A.II) revealed a previously unrecognized genetic division within this highly virulent subspecies. F. tularensis subsp. holarctica appears to have recently spread globally across continents from a single origin, while F. tularensis subsp. tularensis has a long and complex evolutionary history almost exclusively in North America. The sole non-North American type A isolates (Slovakian) were closely related to the SCHU S4 strain. Significant linkage disequilibrium was detected among VNTR loci of F. tularensis consistent with a clonal population structure. Overall, this work greatly augments the study of tularemia ecology and epidemiology, while providing a framework for future forensic analysis of F. tularensis isolates.

A.J. and J.F. contributed equally to this work.

This article has been cited by other articles:

• Oyston, P. C. F. (2008). Francisella tularensis: unravelling the secrets of an intracellular pathogen. J Med Microbiol 57: 921-930 [Abstract] [Full Text]  
• Ludu, J. S., de Bruin, O. M., Duplantis, B. N., Schmerk, C. L., Chou, A. Y., Elkins, K. L., Nano, F. E. (2008). The Francisella Pathogenicity Island Protein PdpD Is Required for Full Virulence and Associates with Homologues of the Type VI Secretion System. J. Bacteriol. 190: 4584-4595 [Abstract] [Full Text]  
• Lembo, A., Pelletier, M., Iyer, R., Timko, M., Dudda, J. C., West, T. E., Wilson, C. B., Hajjar, A. M., Skerrett, S. J. (2008). Administration of a Synthetic TLR4 Agonist Protects Mice from Pneumonic Tularemia. J. Immunol. 180: 7574-7581 [Abstract] [Full Text]  
• La Scola, B., Elkarkouri, K., Li, W., Wahab, T., Fournous, G., Rolain, J.-M., Biswas, S., Drancourt, M., Robert, C., Audic, S., Lofdahl, S., Raoult, D. (2008). Rapid comparative genomic analysis for clinical microbiology: The Francisella tularensis paradigm. Genome Res 18: 742-750 [Abstract] [Full Text]  
• Eisen, R. J., Mead, P. S., Meyer, A. M., Pfaff, L. E., Bradley, K. K., Eisen, L. (2008). Ecoepidemiology of Tularemia in the Southcentral United States. Am J Trop Med Hyg 78: 586-594 [Abstract] [Full Text]  
• Gorge, O., Lopez, S., Hilaire, V., Lisanti, O., Ramisse, V., Vergnaud, G. (2008). Selection and Validation of a Multilocus Variable-Number Tandem-Repeat Analysis Panel for Typing Shigella spp.. J. Clin. Microbiol. 46: 1026-1036 [Abstract] [Full Text]  
• Dempsey, M. P., Dobson, M., Zhang, C., Zhang, M., Lion, C., Gutierrez-Martin, C. B., Iwen, P. C., Fey, P. D., Olson, M. E., Niemeyer, D., Francesconi, S., Crawford, R., Stanley, M., Rhodes, J., Wagner, D. M., Vogler, A. J., Birdsell, D., Keim, P., Johansson, A., Hinrichs, S. H., Benson, A. K. (2007). Genomic Deletion Marking an Emerging Subclone of Francisella tularensis subsp. holarctica in France and the Iberian Peninsula. Appl. Environ. Microbiol. 73: 7465-7470 [Abstract] [Full Text]  
• Guina, T., Radulovic, D., Bahrami, A. J., Bolton, D. L., Rohmer, L., Jones-Isaac, K. A., Chen, J., Gallagher, L. A., Gallis, B., Ryu, S., Taylor, G. K., Brittnacher, M. J., Manoil, C., Goodlett, D. R. (2007). MglA Regulates Francisella tularensis subsp. novicida (Francisella novicida) Response to Starvation and Oxidative Stress. J. Bacteriol. 189: 6580-6586 [Abstract] [Full Text]  
• Karpathy, S. E., Dasch, G. A., Eremeeva, M. E. (2007). Molecular Typing of Isolates of Rickettsia rickettsii by Use of DNA Sequencing of Variable Intergenic Regions. J. Clin. Microbiol. 45: 2545-2553 [Abstract] [Full Text]  
• Vishnoi, A., Roy, R., Bhattacharya, A. (2007). Comparative analysis of bacterial genomes: identification of divergent regions in mycobacterial strains using an anchor-based approach. Nucleic Acids Res 35: 3654-3667 [Abstract] [Full Text]  
• Monteil, M., Durand, B., Bouchouicha, R., Petit, E., Chomel, B., Arvand, M., Boulouis, H.-J., Haddad, N. (2007). Development of discriminatory multiple-locus variable number tandem repeat analysis for Bartonella henselae. Microbiology 153: 1141-1148 [Abstract] [Full Text]  
• Boxrud, D., Pederson-Gulrud, K., Wotton, J., Medus, C., Lyszkowicz, E., Besser, J., Bartkus, J. M. (2007). Comparison of Multiple-Locus Variable-Number Tandem Repeat Analysis, Pulsed-Field Gel Electrophoresis, and Phage Typing for Subtype Analysis of Salmonella enterica Serotype Enteritidis. J. Clin. Microbiol. 45: 536-543 [Abstract] [Full Text]  
• Pasqualotto, A. C., Denning, D. W., Anderson, M. J. (2007). A Cautionary Tale: Lack of Consistency in Allele Sizes between Two Laboratories for a Published Multilocus Microsatellite Typing System. J. Clin. Microbiol. 45: 522-528 [Abstract] [Full Text]  
• Wobeser, G., Ngeleka, M., Appleyard, G., Bryden, L., Mulvey, M. R. (2007). TULAREMIA IN DEER MICE (PEROMYSCUS MANICULATUS) DURING A POPULATION IRRUPTION IN SASKATCHEWAN, CANADA. J Wildl Dis 43: 23-31 [Abstract] [Full Text]  
• Hajjar, A. M., Harvey, M. D., Shaffer, S. A., Goodlett, D. R., Sjostedt, A., Edebro, H., Forsman, M., Bystrom, M., Pelletier, M., Wilson, C. B., Miller, S. I., Skerrett, S. J., Ernst, R. K. (2006). Lack of In Vitro and In Vivo Recognition of Francisella tularensis Subspecies Lipopolysaccharide by Toll-Like Receptors. Infect. Immun. 74: 6730-6738 [Abstract] [Full Text]  
• Rohmer, L., Brittnacher, M., Svensson, K., Buckley, D., Haugen, E., Zhou, Y., Chang, J., Levy, R., Hayden, H., Forsman, M., Olson, M., Johansson, A., Kaul, R., Miller, S. I. (2006). Potential Source of Francisella tularensis Live Vaccine Strain Attenuation Determined by Genome Comparison. Infect. Immun. 74: 6895-6906 [Abstract] [Full Text]  
• LoVullo, E. D., Sherrill, L. A., Perez, L. L., Pavelka, M. S. Jr (2006). Genetic tools for highly pathogenic Francisella tularensis subsp. tularensis.. Microbiology 152: 3425-3435 [Abstract] [Full Text]  
• Petrosino, J. F., Xiang, Q., Karpathy, S. E., Jiang, H., Yerrapragada, S., Liu, Y., Gioia, J., Hemphill, L., Gonzalez, A., Raghavan, T. M., Uzman, A., Fox, G. E., Highlander, S., Reichard, M., Morton, R. J., Clinkenbeard, K. D., Weinstock, G. M. (2006). Chromosome Rearrangement and Diversification of Francisella tularensis Revealed by the Type B (OSU18) Genome Sequence.. J. Bacteriol. 188: 6977-6985 [Abstract] [Full Text]  
• Nix, E. B., Cheung, K. K. M., Wang, D., Zhang, N., Burke, R. D., Nano, F. E. (2006). Virulence of Francisella spp. in Chicken Embryos.. Infect. Immun. 74: 4809-4816 [Abstract] [Full Text]  
• Dempsey, M. P., Nietfeldt, J., Ravel, J., Hinrichs, S., Crawford, R., Benson, A. K. (2006). Paired-End Sequence Mapping Detects Extensive Genomic Rearrangement and Translocation during Divergence of Francisella tularensis subsp. tularensis and Francisella tularensis subsp. holarctica Populations.. J. Bacteriol. 188: 5904-5914 [Abstract] [Full Text]  
• Nubel, U., Reissbrodt, R., Weller, A., Grunow, R., Porsch-Ozcurumez, M., Tomaso, H., Hofer, E., Splettstoesser, W., Finke, E.-J., Tschape, H., Witte, W. (2006). Population Structure of Francisella tularensis.. J. Bacteriol. 188: 5319-5324 [Abstract] [Full Text]  
• Whatmore, A. M., Shankster, S. J., Perrett, L. L., Murphy, T. J., Brew, S. D., Thirlwall, R. E., Cutler, S. J., MacMillan, A. P. (2006). Identification and Characterization of Variable-Number Tandem-Repeat Markers for Typing of Brucella spp.. J. Clin. Microbiol. 44: 1982-1993 [Abstract] [Full Text]  
• Chandler, D. P., Alferov, O., Chernov, B., Daly, D. S., Golova, J., Perov, A., Protic, M., Robison, R., Schipma, M., White, A., Willse, A. (2006). Diagnostic Oligonucleotide Microarray Fingerprinting of Bacillus Isolates. J. Clin. Microbiol. 44: 244-250 [Abstract] [Full Text]  
• Bystrom, M., Bocher, S., Magnusson, A., Prag, J., Johansson, A. (2005). Tularemia in Denmark: Identification of a Francisella tularensis subsp. holarctica Strain by Real-Time PCR and High-Resolution Typing by Multiple-Locus Variable-Number Tandem Repeat Analysis. J. Clin. Microbiol. 43: 5355-5358 [Abstract] [Full Text]  
• Barns, S. M., Grow, C. C., Okinaka, R. T., Keim, P., Kuske, C. R. (2005). Detection of Diverse New Francisella-Like Bacteria in Environmental Samples. Appl. Environ. Microbiol. 71: 5494-5500 [Abstract] [Full Text]  
• Francois, P., Huyghe, A., Charbonnier, Y., Bento, M., Herzig, S., Topolski, I., Fleury, B., Lew, D., Vaudaux, P., Harbarth, S., van Leeuwen, W., van Belkum, A., Blanc, D. S., Pittet, D., Schrenzel, J. (2005). Use of an Automated Multiple-Locus, Variable-Number Tandem Repeat-Based Method for Rapid and High-Throughput Genotyping of Staphylococcus aureus Isolates. J. Clin. Microbiol. 43: 3346-3355 [Abstract] [Full Text]  
• Svensson, K., Larsson, P., Johansson, D., Bystrom, M., Forsman, M., Johansson, A. (2005). Evolution of Subspecies of Francisella tularensis. J. Bacteriol. 187: 3903-3908 [Abstract] [Full Text]