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Journal of Clinical Microbiology, March 2008, p. 1026-1036, Vol. 46, No. 3
0095-1137/08/$08.00+0     doi:10.1128/JCM.02027-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Selection and Validation of a Multilocus Variable-Number Tandem-Repeat Analysis Panel for Typing Shigella spp.{triangledown} ,{dagger}

Olivier Gorgé,1* Stéphanie Lopez,1 Valérie Hilaire,1 Olivier Lisanti,1 Vincent Ramisse,1 and Gilles Vergnaud1,2

Department of Analytical Microbiology, Centre d'Etudes du Bouchet, BP3, F-91710 Vert-le-Petit, France,1 Université Paris-Sud, Institut de Génétique et Microbiologie, and CNRS, Orsay, F-91405, France2

Received 17 October 2007/ Returned for modification 17 December 2007/ Accepted 14 January 2008

The Shigella genus has historically been separated into four species, based on biochemical assays. The classification within each species relies on serotyping. Recently, genome sequencing and DNA assays, in particular the multilocus sequence typing (MLST) approach, greatly improved the current knowledge of the origin and phylogenetic evolution of Shigella spp. The Shigella and Escherichia genera are now considered to belong to a unique genomospecies. Multilocus variable-number tandem-repeat (VNTR) analysis (MLVA) provides valuable polymorphic markers for genotyping and performing phylogenetic analyses of highly homogeneous bacterial pathogens. Here, we assess the capability of MLVA for Shigella typing. Thirty-two potentially polymorphic VNTRs were selected by analyzing in silico five Shigella genomic sequences and subsequently evaluated. Eventually, a panel of 15 VNTRs was selected (i.e., MLVA15 analysis). MLVA15 analysis of 78 strains or genome sequences of Shigella spp. and 11 strains or genome sequences of Escherichia coli distinguished 83 genotypes. Shigella population cluster analysis gave consistent results compared to MLST. MLVA15 analysis showed capabilities for E. coli typing, providing classification among pathogenic and nonpathogenic E. coli strains included in the study. The resulting data can be queried on our genotyping webpage (http://mlva.u-psud.fr). The MLVA15 assay is rapid, highly discriminatory, and reproducible for Shigella and Escherichia strains, suggesting that it could significantly contribute to epidemiological trace-back analysis of Shigella infections and pathogenic Escherichia outbreaks. Typing was performed on strains obtained mostly from collections. Further studies should include strains of much more diverse origins, including all pathogenic E. coli types.

* Corresponding author. Mailing address: Department of Analytical Microbiology, Centre d'Etudes du Bouchet, BP3, F-91710 Vert-le-Petit, France. Phone: 33-1-69-90-82-23. Fax: 33-1-69-90-83-35. E-mail: olivier.gorge{at}dga.defense.gouv.fr

{triangledown} Published ahead of print on 23 January 2008.

{dagger} Supplemental material for this article may be found at http://jcm.asm.org/.

Journal of Clinical Microbiology, March 2008, p. 1026-1036, Vol. 46, No. 3
0095-1137/08/$08.00+0     doi:10.1128/JCM.02027-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Chiou, C.-S., Watanabe, H., Wang, Y.-W., Wang, W.-L., Terajima, J., Thong, K.-L., Phung, D. C., Tung, S. K. (2009). Utility of Multilocus Variable-Number Tandem-Repeat Analysis as a Molecular Tool for Phylogenetic Analysis of Shigella sonnei. J. Clin. Microbiol. 47: 1149-1154 [Abstract] [Full Text]  


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