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Journal of Clinical Microbiology, March 2007, p. 691-697, Vol. 45, No. 3
0095-1137/07/$08.00+0     doi:10.1128/JCM.01393-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Assessment of an Optimized Mycobacterial Interspersed Repetitive- Unit-Variable-Number Tandem-Repeat Typing System Combined with Spoligotyping for Population-Based Molecular Epidemiology Studies of Tuberculosis{triangledown} ,{ddagger}

Mara Cardoso Oelemann,1,2,§ Roland Diel,3 Vincent Vatin,2,4 Walter Haas,5 Sabine Rüsch-Gerdes,6 Camille Locht,1,2 Stefan Niemann,6,{dagger} and Philip Supply1,2*,{dagger}

INSERM, U629,1 Institut Pasteur de Lille, Lille, France,2 School of Public Health, University of Düsseldorf, Düsseldorf, Germany,3 CNRS UMR 8090, Lille, France,4 Department for Infectious Disease Epidemiology, Robert Koch-Institut, Berlin, Germany,5 Forschungszentrum Borstel, National Reference Center for Mycobacteria, Borstel, Germany6

Received 6 July 2006/ Returned for modification 13 October 2006/ Accepted 13 December 2006

An optimized set of 24 mycobacterial interspersed repetitive-unit-variable-number tandem-repeat (MIRU-VNTR) loci, including a discriminatory subset of 15 loci, has recently been defined for the typing of Mycobacterium tuberculosis. Here, we evaluated the performances of this MIRU-VNTR typing system in combination with spoligotyping for the detection of transmission chains in a population-based study comprising 91% of culture-confirmed tuberculosis patients reported in 2003 in Hamburg, Germany. Of the 154 isolates investigated, more than 90% had high IS6110 copy numbers (≥6). IS6110 restriction fragment length polymorphism (RFLP) typing resulted in 13 clusters, 5 of which had a confirmed epidemiological link. All five, as well as six of the eight IS6110 clusters with no identified epidemiological link, were perfectly matched by MIRU-VNTR typing with the 24 loci. Two IS6110 clusters were split by differences into 6 to 12 MIRU-VNTR loci, clearly supporting the absence of a link, as judged by contact tracing data. In contrast, only one MIRU-VNTR cluster, grouping what were probably epidemiologically unlinked isolates, was split by IS6110 RFLP. However, these isolates were also distinguished by spoligotyping. Both the optimized 24-locus and 15-locus sets thus showed a comparable to slightly better predictive value, especially when combined with spoligotyping, than the current gold standard IS6110 RFLP for the study of tuberculosis transmission in Hamburg. Because the epidemiological characteristics of this setting are similar to those of many developed countries, these results support the wide applicability of this real-time genotyping approach for population-based studies of M. tuberculosis transmission.

* Corresponding author. Mailing address: Laboratoire de Mécanismes Moléculaires de la Pathogenèse Bactérienne, INSERM U629, Institut Pasteur de Lille, 1, Rue du Prof. Calmette F-59019 Lille Cedex, France. Phone: (33) 320-871154. Fax: (33) 320-871158. E-mail: philip.supply{at}pasteur-lille.fr.

{triangledown} Published ahead of print on 27 December 2006.

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

§ Present address: Laboratório de Biologia Molecular Aplicada a Micobactérias, Departamento de Micobacterioses, Instituto Oswaldo Cruz-Fiocruz, Rio de Janeiro, RJ, Brazil.

{dagger} S.N. and P.S. contributed equally to this study.

Journal of Clinical Microbiology, March 2007, p. 691-697, Vol. 45, No. 3
0095-1137/07/$08.00+0     doi:10.1128/JCM.01393-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.



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