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Journal of Clinical Microbiology, January 2000, p. 301-308, Vol. 38, No. 1
0095-1137/0/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Differentiation of Phylogenetically Related Slowly Growing Mycobacteria by Their gyrB Sequences

Hiroaki Kasai,1,* Takayuki Ezaki,2 and Shigeaki Harayama1

Marine Biotechnology Institute, Kamaishi Laboratories, 3-75-1 Heita, Kamaishi, Iwate 026-0001,1 and Department of Microbiology, Gifu University School of Medicine, Gifu, Gifu 500-8705,2 Japan

Received 27 May 1999/Returned for modification 18 August 1999/Accepted 4 October 1999

The conventional methods for identifying mycobacterial species are based on their phenotypic characterization. Since some problematic species are slow growers, their taxonomy takes several weeks or months to identify. The ribosomal DNA (rDNA) sequence-based identification strategy has been adopted to solve this problem. More recently, the gyrB sequences have been shown to be useful phylogenetic markers for the identification of species. We determined the gyrB sequences of 43 slowly growing strains belonging to 15 species in the genus Mycobacterium. The frequencies of base substitutions in the gyrB sequences were comparable to those in the 16S-23S rDNA internal transcribed spacer (ITS) sequences. The ITS sequences of four species belonging to the M. tuberculosis complex (M. tuberculosis, M. bovis, M. africanum, and M. microti) were 100% identical, while four synonymous substitutions were found in the gyrB sequences of these strains. Based on the differences found in the gyrB sequences, we developed PCR and PCR-restriction fragment length polymorphism methods to discriminate these species.

* Corresponding author. Mailing address: Marine Biotechnology Institute, Kamaishi Laboratories, 3-75-1 Heita, Kamaishi, Iwate 026-0001, Japan. Phone: 81-193-26-6544. Fax: 81-193-26-6584/6592. E-mail: hiroaki.kasai{at}kamaishi.mbio.co.jp.

Journal of Clinical Microbiology, January 2000, p. 301-308, Vol. 38, No. 1
0095-1137/0/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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