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Journal of Clinical Microbiology, February 1998, p. 499-505, Vol. 36, No. 2
0095-1137/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Separation among Species of Mycobacterium terrae Complex by Lipid Analyses: Comparison with Biochemical Tests and 16S rRNA Sequencing

Pirjo Torkko,1,* Merja Suutari,1 Sini Suomalainen,2 Lars Paulin,2 Lennart Larsson,3 and Marja-Leena Katila4

Laboratory of Environmental Microbiology, National Public Health Institute, Fin-70701 Kuopio,1 Institute of Biotechnology, University of Helsinki, Fin-00014 Helsinki University,2 and Department of Clinical Microbiology, Kuopio University Hospital, Fin-70211 Kuopio,4 Finland, and Department of Medical Microbiology, Lund University, S-22362 Lund, Sweden3

Received 23 June 1997/Returned for modification 11 August 1997/Accepted 10 November 1997

Fatty acids, alcohols, and mycolic acid cleavage products were determined for 13 ATCC strains and 24 clinical isolates, which were initially identified by biochemical and growth characteristics as the Mycobacterium terrae complex. The clinical isolates were also analyzed by partial sequencing of the 16S rRNA gene, which divided them into five genetic entities, M. triviale (three strains), M. terrae (four strains), M. nonchromogenicum sensu stricto (seven strains), Mycobacterium sp. strain MCRO 6 (seven strains), and Mycobacterium sp. strain 31958 (one strain). After acidic methanolysis, secondary alcohols were a characteristic feature in all members of the M. terrae complex but M. triviale. In addition to the prominent secondary alcohols, 2-octadecanol and 2-eicosanol, two previously unidentified alcohols, 2-(8,15-dimethyl)docosenol and 2-(8,17-dimethyl)tetracosenol, were detected in M. nonchromogenicum, Mycobacterium sp. strain MCRO 6, and Mycobacterium sp. strain 31958. Only 2-(8,17-dimethyl)tetracosenol was detected in trace amounts in M. terrae. Genetic differences were associated with differences in phenotypic characteristics, including growth at 42°C and pyrazinamidase production. Based on fatty acid and alcohol composition and biochemical and genetic characteristics, M. nonchromogenicum and Mycobacterium sp. strains MCRO 6 and 31958 were found to be a closely related group, named the M. nonchromogenicum complex. Detected genetic variations associated with phenotypic characteristics may indicate further species separation of this complex. In conclusion, the results of gas-liquid chromatography fatty acid analysis, combined with those of a Tween 80 test, enable identification of the species of the M. terrae complex and their separation from other nonpigmented slowly growing mycobacteria.

* Corresponding author. Mailing address: Dept. Environmental Microbiology, National Public Health Institute, P.O. Box 95, FIN-70701 Kuopio, Finland. Fax: 358-17-201155. E-mail: Pirjo.Torkko{at}ktl.fi.

Journal of Clinical Microbiology, February 1998, p. 499-505, Vol. 36, No. 2
0095-1137/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.


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