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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.

Pyrosequencing assay for rapid identification of Mycobacterium tuberculosis complex species