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Journal of Clinical Microbiology, March 1999, p. 852-857, Vol. 37, No. 3
0095-1137/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

hsp65 Sequencing for Identification of Rapidly Growing Mycobacteria

H. Ringuet,1 C. Akoua-Koffi,1 S. Honore,1 A. Varnerot,2 V. Vincent,2 P. Berche,1 J. L. Gaillard,1 and C. Pierre-Audigier1,*

Service de Microbiologie, Hôpital Necker-Enfants Malades, 75015 Paris,1 and Centre National de Référence des Mycobactéries, Institut Pasteur, 75724 Paris,2 France

Received 20 May 1998/Returned for modification 25 June 1998/Accepted 19 November 1998

Partial sequencing of the hsp65 gene was used for the identification of rapidly growing mycobacteria (RGM). A 441-bp fragment (A. Telenti, F. Marchesi, M. Balz, F. Bally, E. Böttger, and T. Bodmer, J. Clin. Microbiol. 31:175-178, 1993) was amplified and sequenced by an automated fluorescence-based method involving capillary electrophoresis. Type strains of 10 RGM species were first studied. Each species had a unique nucleotide sequence, distinguishing it clearly from the other species. A panel of strains from the four main RGM species responsible for human infections, Mycobacterium abscessus, Mycobacterium chelonae, Mycobacterium fortuitum, and Mycobacterium peregrinum, was also studied. There were few sequence differences within each of these species (<2% of bases were different from the type strain sequence), and they had no effect on species assignment. hsp65 sequencing unambiguously differentiated M. chelonae and M. abscessus, two species difficult to identify by classical methods and 16S rRNA gene sequencing. The devised procedure is a rapid and reliable tool for the identification of RGM species.

* Corresponding author. Mailing address: Laboratoire de Microbiologie, Hôpital Necker-Enfants Malades, 149 rue de Sèvres, 75743 Paris Cedex 15, France. Phone: (33) 01 44 49 49 61. Fax: (33) 01 44 49 49 60. E-mail: berche{at}necker.fr.

Journal of Clinical Microbiology, March 1999, p. 852-857, Vol. 37, No. 3
0095-1137/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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