Affiliation:
1. National Reference Centre for Mycobacteriology, Bureau of Microbiology, Health Canada,1 and
2. Department of Clinical Microbiology, Health Sciences Centre,2 Winnipeg, Manitoba, Canada
Abstract
ABSTRACT
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.
Publisher
American Society for Microbiology
Cited by
85 articles.
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