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Journal of Clinical Microbiology, February 2006, p. 433-440, Vol. 44, No. 2
0095-1137/06/$08.00+0     doi:10.1128/JCM.44.2.433-440.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Sequencing of hsp65 Distinguishes among Subsets of the Mycobacterium avium Complex

Christine Y. Turenne,1 Makeda Semret,1 Debby V. Cousins,2 Desmond M. Collins,3 and Marcel A. Behr1*

McGill University Health Centre, Montreal, Quebec, Canada H3G 1A4,1 Australian Reference Laboratory for Bovine Tuberculosis, Department of Agriculture, South Perth, Western Australia, Australia 6151,2 AgResearch, Wallaceville Animal Research Centre, Upper Hutt, New Zealand3

Received 8 September 2005/ Returned for modification 2 November 2005/ Accepted 16 November 2005

The Mycobacterium avium complex consists of epidemiologically distinct subsets. The classification of these subsets is complicated by a number of factors, including the ambiguous results obtained with phenotypic and genetic assays and the recent appreciation that human and avian strains appear to be distinct. In previous work, sequencing based on a 441-bp portion of the hsp65 gene has proven to efficiently classify isolates within the Mycobacterium genus but provides low resolution for distinguishing among members of the M. avium complex. Therefore, in this study, we have targeted the more variable 3' region of the hsp65 gene to determine whether it can effectively discriminate M. avium complex isolates at the levels of species and subspecies. Primers designed for this target consistently generated amplicons for all organisms classified as M. avium complex. Sequences obtained indicate that M. intracellulare is genetically divergent from M. avium organisms, and distinct sequevars were obtained for M. avium subsets, including M. avium subsp. avium (bird type), M. avium subsp. hominissuis, and M. avium subsp. paratuberculosis. In addition, sequence differences served to distinguish bovine from ovine strains of M. avium subsp. paratuberculosis. A unique profile for M. avium subsp. silvaticum was not obtained. These results indicate that sequencing the 3' region of the hsp65 gene can simply and unambiguously distinguish species and subspecies of the M. avium complex.

* Corresponding author. Mailing address: Division of Infectious Diseases and Medical Microbiology, A5-156, Montreal General Hospital, 1650 Cedar Avenue, Montreal, QC H3G 1A4 Canada. Phone: 514 934 1934, ext. 42815. Fax: 514 934 8423. E-mail: marcel.behr{at}mcgill.ca.

Journal of Clinical Microbiology, February 2006, p. 433-440, Vol. 44, No. 2
0095-1137/06/$08.00+0     doi:10.1128/JCM.44.2.433-440.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.



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