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Journal of Clinical Microbiology, April 2003, p. 1656-1663, Vol. 41, No. 4
0095-1137/03/$08.00+0     DOI: 10.1128/JCM.41.4.1656-1663.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Heterogeneity of RNA Polymerase Gene (rpoB) Sequences of Mycobacterium gordonae Clinical Isolates Identified with a DNA Probe Kit and by Conventional Methods

Saotomo Itoh, Yuko Kazumi, Chiyoji Abe, and Mitsuyoshi Takahashi*

Bacteriology Division, The Research Institute of Tuberculosis, Japan Antituberculosis Association, Kiyose-shi, Tokyo 204-8533, Japan

Received 3 July 2002/ Returned for modification 23 August 2002/ Accepted 2 January 2003

In a previous study, we have evaluated genetic identification by using the rpoB gene, which was recently introduced by Kim et al. (J. Clin. Microbiol. 39:2102-2109, 2001; J. Clin. Microbiol. 37:1714-1720, 1999). In this process, we examined the rpoB gene heterogeneity of clinical isolates identified as Mycobacterium gordonae with the conventional biological and biochemical tests and/or a commercially available DNA probe kit. Sequencing of the rpoB gene of 34 clinical isolates revealed that M. gordonae clinical isolates were classified into four major clusters (A, B, C, and D). Interestingly, organisms belonging to cluster D (15 isolates) did not hybridize with M. gordonae ATCC 14470 and specifically possessed urease activity. Therefore, it could be considered to be a novel mycobacterium. The identification of M. gordonae is known to have ambiguous results sometimes. On the other hand, identification of clinical isolates seems to be inconvenient and unsuitable because of a more than 99% 16S rRNA gene similarity value between clusters. These findings suggest that the existence of M. gordonae-like mycobacteria that share similar biochemical and biological characteristics with the 16S rRNA gene of an M. gordonae type strain but less similarity at the genomic DNA level may have complicated the identification of M. gordonae in many laboratories. Furthermore, compared with hsp65 PCR restriction analysis (PRA), rpoB PRA would have the advantage of producing no ambiguous results because of the intracluster homogeneity of the rpoB gene. In this case, rpoB would provide clearer results than hsp65, even if PRA analysis was used. We demonstrated that these M. gordonae-like mycobacteria were easily distinguished by PRA of the rpoB sequence. Additionally, the significance of this M. gordonae-like cluster may help to establish the comparison between the M. gordonae isolates from a clinical specimen and an infectious process in a given patient and to determine the true incidence of infection with this microorganism.

* Corresponding author. Mailing address: Research Institute of Tuberculosis, Japan Antituberculosis Association, Kiyose-shi, Tokyo 204-8533, Japan. Phone: 81-424-93-5773. Fax: 81-424-92-4600. E-mail: tak{at}jata.or.jp.

Journal of Clinical Microbiology, April 2003, p. 1656-1663, Vol. 41, No. 4
0095-1137/03/$08.00+0     DOI: 10.1128/JCM.41.4.1656-1663.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • McNabb, A., Eisler, D., Adie, K., Amos, M., Rodrigues, M., Stephens, G., Black, W. A., Isaac-Renton, J. (2004). Assessment of Partial Sequencing of the 65-Kilodalton Heat Shock Protein Gene (hsp65) for Routine Identification of Mycobacterium Species Isolated from Clinical Sources. J. Clin. Microbiol. 42: 3000-3011 [Abstract] [Full Text]  


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