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Journal of Clinical Microbiology, August 2000, p. 2846-2852, Vol. 38, No. 8
0095-1137/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Evaluation of recA Sequences for Identification of Mycobacterium Species

Kym S. Blackwood,1 Cheng He,2 James Gunton,1 Christine Y. Turenne,1,* Joyce Wolfe,1,2 and Amin M. Kabani1,2

National Reference Centre for Mycobacteriology, Bureau of Microbiology, Health Canada,1 and Department of Clinical Microbiology, Health Sciences Centre,2 Winnipeg, Manitoba, Canada

Received 3 April 2000/Returned for modification 1 May 2000/Accepted 29 May 2000

16S rRNA sequence data have been used to provide a molecular basis for an accurate system for identification of members of the genus Mycobacterium. Previous studies have shown that Mycobacterium species demonstrate high levels (>94%) of 16S rRNA sequence similarity and that this method cannot differentiate between all species, i.e., M. gastri and M. kansasii. In the present study, we have used the recA gene as an alternative sequencing target in order to complement 16S rRNA sequence-based genetic identification. The recA genes of 30 Mycobacterium species were amplified by PCR, sequenced, and compared with the published recA sequences of M. tuberculosis, M. smegmatis, and M. leprae available from GenBank. By recA sequencing the species showed a lower degree of interspecies similarity than they did by 16S rRNA gene sequence analysis, ranging from 96.2% between M. gastri and M. kansasii to 75.7% between M. aurum and M. leprae. Exceptions to this were members of the M. 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 new Mycobacterium species that contain a protein intron in their recA genes, similar to M. tuberculosis and M. leprae. We propose that recA gene sequencing offers a complementary method to 16S rRNA gene sequencing for the accurate identification of the Mycobacterium species.

* Corresponding author. Mailing address: National Reference Centre for Mycobacteriology, Canadian Science Centre for Human and Animal Health, 1015 Arlington St., Winnipeg, Manitoba, Canada, R3E 3R2. Phone: (204) 789-6081. Fax: (204) 789-2036. E-mail: cturenne{at}hc-sc.gc.ca.

Journal of Clinical Microbiology, August 2000, p. 2846-2852, Vol. 38, No. 8
0095-1137/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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