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Journal of Clinical Microbiology, August 2000, p. 2966-2971, Vol. 38, No. 8
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

Hyeyoung Lee,1 Hee-Jung Park,1 Sang-Nae Cho,2 Gill-Han Bai,1 and Sang-Jae Kim1,*

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.

* 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.

Journal of Clinical Microbiology, August 2000, p. 2966-2971, Vol. 38, No. 8
0095-1137/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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