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16S rRNA sequence data have been used to provide a molecular basis for an accurate system for identification of members of the genusMycobacterium . Previous studies have shown thatMycobacterium species demonstrate high levels (&gt;94%) of 16S rRNA sequence similarity and that this method cannot differentiate between all species, i.e.,M. gastri andM. kansasii . In the present study, we have used therecA gene as an alternative sequencing target in order to complement 16S rRNA sequence-based genetic identification. TherecA genes of 30Mycobacterium species were amplified by PCR, sequenced, and compared with the publishedrecA sequences ofM. tuberculosis ,M. smegmatis , andM. leprae available from GenBank. ByrecA sequencing the species showed a lower degree of interspecies similarity than they did by 16S rRNA gene sequence analysis, ranging from 96.2% betweenM. gastri andM. kansasii to 75.7% betweenM. aurum andM. leprae . Exceptions to this were members of theM. tuberculosis complex, which were identical. Two strains of each of 27 species were tested, and the intraspecies similarity ranged from 98.7 to 100%. In addition, we identified newMycobacterium species that contain a protein intron in theirrecA genes, similar toM. tuberculosis andM. leprae . We propose thatrecA gene sequencing offers a complementary method to 16S rRNA gene sequencing for the accurate identification of theMycobacterium species.

Evaluation of recA Sequences for Identification of Mycobacterium Species