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Journal of Clinical Microbiology, June 2006, p. 1951-1962, Vol. 44, No. 6
0095-1137/06/$08.00+0     doi:10.1128/JCM.01775-05
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Evaluation of the Epidemiological Relevance of Variable-Number Tandem-Repeat Genotyping of Mycobacterium bovis and Comparison of the Method with IS6110 Restriction Fragment Length Polymorphism Analysis and Spoligotyping

Caroline Allix,¹ Karl Walravens,² Claude Saegerman,^3, Jacques Godfroid,^2, Philip Supply,^4,¶ and Maryse Fauville-Dufaux^1*,¶

Institut Pasteur de Bruxelles, Laboratoire Tuberculose et Mycobactéries, rue Engeland 642, 1180 Bruxelles, Belgium,¹ Centre d'Etude et de Recherches Vétérinaires et Agrochimiques (CERVA-CODA), Section Maladies Bactériennes et Immunologie, Groeselenberg 99, 1180 Bruxelles, Belgium,² Agence Fédérale pour la Sécurité de la Chaîne Alimentaire, DG Politique de Contrôle, Secrétariat du Comité Scientifique, Avenue Simon Bolivar 30, 1000 Bruxelles, Belgium,³ Institut Pasteur de Lille, Laboratoire des Mécanismes Moléculaires de la Pathogenèse Bactérienne, INSERM U629, 1, rue du Professeur Calmette, BP 245, 59019 Lille Cedex, France⁴

Received 27 August 2005/ Returned for modification 2 December 2005/ Accepted 10 March 2006

Sources of Mycobacterium bovis contamination remain unclear for many cases of animal and human disease. A major limitation is the lack of sufficiently informative or epidemiologically well evaluated molecular methods for typing. Here, we report an evaluation of a high-throughput method based on 29 mycobacterial interspersed repetitive unit-variable-number tandem-repeat (MIRU-VNTR) loci to genotype 127 M. bovis isolates from cattle from 77 different Belgian farms, representative of a nationwide collection obtained from 1995 to 2003. MIRU-VNTR stability was demonstrated by analyzing a series of 74 isolates in total, obtained from different animals from a single farm or from different farms with an identified epidemiological link. The genotyping results and the genotypic diversity (h) were compared with those obtained by IS6110 restriction fragment length polymorphism (RFLP) analysis and spoligotyping. Among 68 isolates with no known epidemiological link, MIRU-VNTR typing discriminated better than either RFLP analysis or spoligotyping, with isolates taken individually (32 versus 16 and 17 genotypes; h = 0.91 versus 0.73 and 0.85, respectively) or in combination (32 versus 28 genotypes; h = 0.91 versus 0.92). Maximal resolution was already achieved with a subset of 9 loci. The observed congruence of the genetic relationships based on IS6110 RFLP analysis, spoligotyping, and MIRU-VNTR markers is consistent with a clonal population structure of M. bovis. These results support MIRU-VNTR typing as a convenient and discriminatory technique for analysis of the population structure of M. bovis in much greater detail and for addressing some still unresolved issues in the epidemiology of the pathogen.

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

Present address: Department of Infectious and Parasitic Diseases, Epidemiology and Risk Analysis, Faculty of Veterinary Medicine, University of Liège, Boulevard de Colonster, 20, B43b, B-4000 Liege, Belgium.

Present address: University of Pretoria, Faculty of Veterinary Science, 0110 Onderstepoort, South Africa.

^¶ P.S. and M.F.-D. contributed equally to the work.

This article has been cited by other articles:

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