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Journal of Clinical Microbiology, December 2002, p. 4561-4566, Vol. 40, No. 12
0095-1137/02/$04.00+0     DOI: 10.1128/JCM.40.12.4561-4566.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Stability of Variable-Number Tandem Repeats of Mycobacterial Interspersed Repetitive Units from 12 Loci in Serial Isolates of Mycobacterium tuberculosis

Evgueni Savine,1 Robin M. Warren,2 Gian D. van der Spuy,2 Nulda Beyers,3 Paul D. van Helden,2 Camille Locht,1 and Philip Supply1*

Laboratoire des Mécanismes Moléculaires de la Pathogenèse Bactérienne, INSERM U447, Institut Pasteur de Lille, F-59019 Lille Cedex, France,1 MRC Centre for Molecular and Cellular Biology, Department of Medical Biochemistry,2 Department of Pediatrics and Child Health, Stellenbosch University, Tygerberg 7505, South Africa3

Received 12 June 2002/ Returned for modification 31 July 2002/ Accepted 2 September 2002

Variable number tandem repeats (VNTRs) of elements named mycobacterial interspersed repetitive units (MIRUs) have previously been identified in 12 minisatellite loci of the Mycobacterium tuberculosis genome. These markers allow reliable high-throughput genotyping of M. tuberculosis and represent a portable approach to global molecular epidemiology of M. tuberculosis. To assess their temporal stability, we genotyped 123 serial isolates, separated by up to 6 years and belonging to a variety of distinct IS6110 restriction fragment length polymorphism (RFLP) families, from 56 patients who had positive sputum cultures. All 12 MIRU VNTR loci were completely identical within the groups of serial isolates in 55 out of 56 groups (98.2%), although 11 pairs of isolates from the same patients with conserved MIRU VNTRs displayed slightly different IS6110 RFLP profiles. In a single case, serial isolates with an unchanged IS6110 RFLP profile showed a change in 1 out of 12 MIRU VNTR loci. These results indicate that MIRU VNTRs are stable over time and therefore are suitable for reliable follow-up of patients chronically infected with tuberculosis over long periods. Moreover, they support MIRU VNTR genotyping as a powerful first-line method followed by subtyping by IS6110 RFLP to define ongoing transmission clusters.

* Corresponding author. Mailing address: Laboratoire de Mécanismes Moléculaires de la Pathogenèse Bactérienne, INSERM U447, Institut Pasteur de Lille, 1, rue du Prof. Calmette, F-59019 Lille Cedex, France. Phone: (33) 320 87 11 54. Fax: (33) 320 87 11 58. E-mail: Philip.Supply{at}pasteur-lille.fr.

Journal of Clinical Microbiology, December 2002, p. 4561-4566, Vol. 40, No. 12
0095-1137/02/$04.00+0     DOI: 10.1128/JCM.40.12.4561-4566.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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