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Journal of Clinical Microbiology, April 2008, p. 1398-1406, Vol. 46, No. 4
0095-1137/08/$08.00+0     doi:10.1128/JCM.02089-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Three-Year Population-Based Evaluation of Standardized Mycobacterial Interspersed Repetitive-Unit-Variable-Number Tandem-Repeat Typing of Mycobacterium tuberculosis{triangledown}

Caroline Allix-Béguec,1,{dagger} Maryse Fauville-Dufaux,1,{ddagger} and Philip Supply2,3,{ddagger}*

Scientific Institute of Public Health, Department Institut Pasteur de Bruxelles, Laboratoire Tuberculose et Mycobacteries, rue Engeland 642, 1180 Bruxelles, Belgium,1 Institut Pasteur de Lille,2 Laboratoire des Mécanismes Moléculaires de la Pathogenèse Bactérienne, INSERM U629-1, rue du Professeur Calmette, BP 245, 59019 Lille Cedex, France3

Received 29 October 2007/ Returned for modification 10 December 2007/ Accepted 22 January 2008

Standardized mycobacterial interspersed repetitive-unit-variable-number tandem repeat (MIRU-VNTR) typing based on 15 and 24 loci recently has been proposed for Mycobacterium tuberculosis genotyping. So far, this optimized system has been assessed in a single, 1-year population-based study performed in Germany (M. C. Oelemann, R. Diel, V. Vatin, W. Haas, S. Rusch-Gerdes, C. Locht, S. Niemann, and P. Supply, J. Clin. Microbiol. 45:691-697, 2007). Here, we evaluated these optimized formats in a much larger population-based study conducted during 39 months in the Brussels capital region of Belgium. Isolates from 807 patients were genotyped. The resolution power, cluster, and lineage identification by the standardized MIRU-VNTR sets were compared to those obtained using standardized IS6110-restriction fragment length polymorphism (RFLP), spoligotyping, and a previous 12-MIRU-VNTR-locus set. On a subset representing 77% of the cases during a 16-month period, a high concordance was observed between unique isolates or strain clusters as defined by standardized MIRU-VNTR and IS6110-RFLP (i.e., more than five IS6110 bands). When extended to the entire population-based collection, the discriminatory subset of 15 loci decreased the strain-clustering rate by almost twofold compared to that of the old 12-locus set. The addition of the nine ancillary MIRU-VNTR loci and/or spoligotyping only slightly further decreased this strain-clustering rate. Familial, social, and/or geographic proximity links were found in 48% of the clusters identified, and well-known risk factors for tuberculosis transmission were identified. Finally, an excellent correspondence was determined between our MIRU-VNTR-spoligotyping strain identifications and external reference strain lineages included in the MIRU-VNTRplus database and identified by, e.g., large sequence polymorphisms. Our results reinforce the proposal of standardized MIRU-VNTR typing as a new reference genotyping method for the epidemiological and phylogenetic screening of M. tuberculosis strains.

* Corresponding author. Mailing address: Molecular Mechanisms of Bacterial Pathogenesis, INSERM U629, Institut de Biologie de Lille/Institut Pasteur de Lille, 1 rue du Professeur Calmette, F-59021 Lille Cedex, France. Phone: (33)320871154. Fax: (33)320871158. E-mail: philip.supply{at}pasteur-lille.fr

{triangledown} Published ahead of print on 30 January 2008.

{dagger} Present address: Genoscreen, Campus de l'Institut Pasteur de Lille, 1 rue du Professeur Calmette, 59000 Lille, France.

{ddagger} These authors contributed equally to this work.

Journal of Clinical Microbiology, April 2008, p. 1398-1406, Vol. 46, No. 4
0095-1137/08/$08.00+0     doi:10.1128/JCM.02089-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.



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