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Journal of Clinical Microbiology, August 2000, p. 2966-2971, Vol. 38, No. 8
Department of Microbiology, Korean Institute
of Tuberculosis, The Korean National Tuberculosis Association,
Seocho-gu, Seoul 137-140,1 and
Department of Microbiology, Yonsei University College of
Medicine, Seodaemoon-gu, Seoul 120-752,2
Korea
Received 18 February 2000/Returned for modification 30 April
2000/Accepted 29 May 2000
PCR-restriction fragment length polymorphism analysis (PRA) using
the novel region of the rpoB gene was developed for rapid and precise identification of mycobacteria to the species level. A
total of 50 mycobacterial reference strains and 3 related bacterial strains were used to amplify the 360-bp region of rpoB, and
the amplified DNAs were subsequently digested with restriction enzymes such as MspI and HaeIII. The results from this
study clearly show that most of the mycobacterial species were easily
differentiated at the species level by this PRA method. In addition,
species with several subtypes, such as Mycobacterium
gordonae, M. kansasii, M. celatum, and
M. fortuitum, were also differentiated by this PRA method.
Subsequently, an algorithm was constructed based on the results, and a
blinded test was carried out with more than 260 clinical isolates that
had been identified on the basis of conventional tests. Comparison of
these two sets of results clearly indicates that this new PRA method
based on the rpoB gene is more simple, more rapid, and more
accurate than conventional procedures for differentiating mycobacterial species.
0095-1137/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.
Species Identification of Mycobacteria by
PCR-Restriction Fragment Length Polymorphism of the
rpoB Gene
*
Corresponding author. Mailing address: Korean Institute
of Tuberculosis, 14 Woomyun-dong, Seocho-Gu, Seoul 137-140, Korea. Phone: 82-2-575-1547. Fax: 82-2-573-1914. E-mail:
sjkim{at}knta.or.kr.
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