Affiliation:
1. Forschungszentrum Borstel, National Reference Center for Mycobacteria, D-23845 Borstel,
1
and
2. Institute for Hygiene & Microbiology, University of Würzburg, D-97080 Würzburg,
2
Germany
Abstract
The discriminatory power of gyrB DNA sequence polymorphisms for differentiation of the species of theMycobacterium tuberculosis complex (MTBC) was evaluated by sequencing and restriction fragment length polymorphism (RFLP) analysis of a 1,020-bp fragment amplified from clinical isolates of M. tuberculosis, Mycobacterium bovis (pyrazinamide [PZA] resistant as well as PZA susceptible), Mycobacterium africanum subtypes I and II, and Mycobacterium microti types vole and llama. We found sequence polymorphisms in four regions described previously and at one additional position. These differences in the gyrB sequences allow an accurate discrimination of M. bovis, M. microti, andM. africanum subtype I. The PZA-susceptible subtypes ofM. bovis shared the M. bovis-specific substitution at position 756 with the PZA-resistant strains, but can be unambiguously differentiated by a characteristic substitution at position 1311. As a drawback, M. tuberculosis and M. africanum subtype II showed an identical gyrBsequence that facilitates discrimination from the other species, but not from each other. A PCR-RFLP technique applying three restriction enzymes could be shown to be a rapid and easy-to-perform tool for the differentiation of the members of the MTBC. Based on these results, we present a clear diagnostic algorithm for the differentiation of species of the MTBC.
Publisher
American Society for Microbiology
Cited by
134 articles.
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