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Open Access Technical Note

Pyrosequencing assay for rapid identification of Mycobacterium tuberculosis complex species

Imen Ben Kahla12, Mireille Henry2, Jalel Boukadida1 and Michel Drancourt2*

Author Affiliations

1 Laboratoire de Microbiologie et d'Immunologie, UR02/SP13, CHU Farhat Hached Sousse, Tunisie

2 URMITE, CNRS UMR6236, IRD198, IFR 48, Institut Méditerranée Infection, Aix-Marseille-Université, Marseille, France

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BMC Research Notes 2011, 4:423  doi:10.1186/1756-0500-4-423

Published: 19 October 2011

Abstract

Background

Identification of the Mycobacterium tuberculosis complex organisms to the species level is important for diagnostic, therapeutic and epidemiologic perspectives. Indeed, isolates are routinely identified as belonging to the M. tuberculosis complex without further discrimination in agreement with the high genomic similarity of the M. tuberculosis complex members and the resulting complex available identification tools.

Findings

We herein develop a pyrosequencing assay analyzing polymorphisms within glpK, pykA and gyrB genes to identify members of the M. tuberculosis complex at the species level. The assay was evaluated with 22 M. tuberculosis, 21 M. bovis, 3 M. caprae, 3 M. microti, 2 M. bovis BCG, 2 M. pinnipedii, 1 M. canettii and 1 M. africanum type I isolates. The resulted pyrograms were consistent with conventional DNA sequencing data and successfully identified all isolates. Additionally, 127 clinical M. tuberculosis complex isolates were analyzed and were unambiguously identified as M. tuberculosis.

Conclusion

We proposed a pyrosequencing-based scheme for the rapid identification of M. tuberculosis complex isolates at the species level. The assay is robust, specific, rapid and can be easily introduced in the routine activity.

Keywords:
Mycobacterium tuberculosis complex; pyrosequencing; identification