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Journal of Clinical Microbiology, August 2005, p. 3704-3712, Vol. 43, No. 8
0095-1137/05/$08.00+0     doi:10.1128/JCM.43.8.3704-3712.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Genomic Polymorphisms for Mycobacterium avium subsp. paratuberculosis Diagnostics

Makeda Semret,1 David C. Alexander,1 Christine Y. Turenne,1 Petra de Haas,2 Pieter Overduin,2 Dick van Soolingen,2 Debby Cousins,3 and Marcel A. Behr1*

McGill University Health Centre, Montreal, Quebec, Canada H3G 1A4,1 National Institute of Public Health and the Environment, 3720BA Bilthoven, The Netherlands,2 Australian Reference Laboratory for Bovine Tuberculosis, Department of Agriculture, South Perth, Western Australia, Australia 61513

Received 16 December 2004/ Returned for modification 13 March 2005/ Accepted 13 May 2005

Mycobacterium avium subsp. paratuberculosis is an emerging pathogen of mammals and is being actively investigated as a possible zoonotic agent. The lack of reliable diagnostic assays has hampered rational assessment of the prevalence of this organism in humans and animals. We have used a comparative genomic approach to reveal genomic differences between M. avium subsp. paratuberculosis and its close relative M. avium subsp. avium, a highly prevalent environmental organism. From computational and DNA microarray-based study of two prototype strains, M. avium subsp. avium strain 104 and M. avium subsp. paratuberculosis strain K10, we have uncovered two types of large sequence polymorphisms (LSPs): those present in the former but missing in the latter (LSPAs) and those only present in the latter (LSPPs). We examined the distribution of 3 LSPAs and 17 LSPPs across a panel of 383 M. avium complex isolates in order to determine their potential utility for the development of accurate diagnostic tests. Our results show that the absence of LSPA8 is 100% specific for the identification of M. avium subsp. paratuberculosis. Of the 17 LSPPs, 10 regions were not specific for M. avium subsp. paratuberculosis while 7 were shown to be highly specific (>98%) and, in some cases, highly sensitive as well (up to 95%). These data highlight the need to evaluate these regions across a diverse panel of clinical and environmental isolates and indicate the LSPs best suited for M. avium subsp. paratuberculosis diagnostics.

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

Journal of Clinical Microbiology, August 2005, p. 3704-3712, Vol. 43, No. 8
0095-1137/05/$08.00+0     doi:10.1128/JCM.43.8.3704-3712.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



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