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Journal of Clinical Microbiology, July 2000, p. 2520-2524, Vol. 38, No. 7
0095-1137/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Molecular Typing of Mycobacterium tuberculosis Based on Variable Number of Tandem DNA Repeats Used Alone and in Association with Spoligotyping

Ingrid Filliol, Severine Ferdinand, Laetitia Negroni, Christophe Sola, and Nalin Rastogi*

Unité de la Tuberculose et des Mycobactéries, Institut Pasteur, F-97165 Pointe-à-Pitre Cedex, Guadeloupe

Received 21 January 2000/Returned for modification 22 March 2000/Accepted 25 April 2000

Fingerprinting based on variable numbers of tandem DNA repeats (VNTR), a recently described methodology, was evaluated for molecular typing of Mycobacterium tuberculosis in an insular setting. In this study, VNTR fingerprinting was used alone or as a second-line test in association with spoligotyping, double-repetitive-element PCR (DRE-PCR), and IS6110 restriction fragment length polymorphism (RFLP) analysis, and the discriminatory power for each method or the combination of methods was compared by calculating the Hunter-Gaston discriminative index (HGI). The results obtained showed that in 6 out of 12 (50%) cases, VNTR-defined clusters were further subdivided by spoligotyping, compared to 7 out of 18 (39%) cases where spoligotyping-defined clusters were further subdivided by VNTR. When used alone, VNTR was the least discriminatory method (HGI = 0.863). Although VNTR was significantly more discriminatory when used in association with spoligotyping (HGI = 0.982), the combination of spoligotyping and DRE-PCR (HGI = 0.992) was still the most efficient among rapid, PCR-based methodologies, giving results comparable to IS6110 RFLP analysis. Nonetheless, VNTR typing may provide additional phylogenetical information that may be helpful to trace the molecular evolution of tubercle bacilli.

* Corresponding author. Mailing address: Unité Tuberculose et Mycobactéries, Institut Pasteur, Morne Jolivière, BP 484, F-97165 Pointe-à-Pitre Cedex, Guadeloupe. Phone: 590-893-881. Fax: 590-893-880. E-mail: rastogi{at}ipagua.gp.

Journal of Clinical Microbiology, July 2000, p. 2520-2524, Vol. 38, No. 7
0095-1137/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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