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Journal of Clinical Microbiology, October 2005, p. 4984-4992, Vol. 43, No. 10
0095-1137/05/$08.00+0     doi:10.1128/JCM.43.10.4984-4992.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Characterization of Mycobacterium caprae Isolates from Europe by Mycobacterial Interspersed Repetitive Unit Genotyping

Wolfgang M. Prodinger,^1* Anita Brandstätter,² Ludmila Naumann,³ Maria Pacciarini,⁴ Tanja Kubica,⁵ Maria Laura Boschiroli,⁶ Alicia Aranaz,⁷ György Nagy,⁸ Zeljko Cvetnic,⁹ Matjaz Ocepek,¹⁰ Artem Skrypnyk,¹¹ Wilfried Erler,¹¹ Stefan Niemann,⁵ Ivo Pavlik,^12, and Irmgard Moser^11,

Department of Hygiene, Microbiology, and Social Medicine,¹ Institute of Legal Medicine, Innsbruck Medical University, Innsbruck, Austria,² Bavarian Health and Food Safety Authority, Oberschleissheim, Germany,³ Istituto Zooprofilattico Sperimentale della Lombardia e dell'Emilia Romagna, Brescia, Italy,⁴ Forschungszentrum Borstel, National Reference Center for Mycobacteria, Borstel, Germany,⁵ Agence Française pour la Sécurité Sanitaire des Aliments, Maisons Alfort, France,⁶ Departamento de Sanidad Animal, Universidad Complutense, Madrid, Spain,⁷ Central Veterinary Institute, Budapest, Hungary,⁸ Croatian Veterinary Institute, Zagreb, Croatia,⁹ Veterinary Faculty, University of Ljubljana, Slovenia,¹⁰ German National Veterinary Reference Laboratory for Tuberculosis, Friedrich-Loeffler-Institut, Jena, Germany,¹¹ Veterinary Research Institute, Brno, Czech Republic,¹²

Received 2 May 2005/ Returned for modification 16 June 2005/ Accepted 17 July 2005

Mycobacterium caprae, a recently defined member of the Mycobacterium tuberculosis complex, causes tuberculosis among animals and, to a limited extent, in humans in several European countries. To characterize M. caprae in comparison with other Mycobacterium tuberculosis complex members and to evaluate genotyping methods for this species, we analyzed 232 M. caprae isolates by mycobacterial interspersed repetitive unit (MIRU) genotyping and by spoligotyping. The isolates originated from 128 distinct epidemiological settings in 10 countries, spanning a period of 25 years. We found 78 different MIRU patterns (53 unique types and 25 clusters with group sizes from 2 to 9) but only 17 spoligotypes, giving Hunter-Gaston discriminatory indices of 0.941 (MIRU typing) and 0.665 (spoligotyping). For a subset of 103 M. caprae isolates derived from outbreaks or endemic foci, MIRU genotyping and IS6110 restriction fragment length polymorphism were compared and shown to provide similar results. MIRU loci 4, 26, and 31 were most discriminant in M. caprae, followed by loci 10 and 16, a combination which is different than those reported to discriminate M. bovis best. M. caprae MIRU patterns together with published data were used for phylogenetic inference analysis employing the neighbor-joining method. M. caprae isolates were grouped together, closely related to the branches of classical M. bovis, M. pinnipedii, M. microti, and ancestral M. tuberculosis, but apart from modern M. tuberculosis. The analysis did not reflect geographic patterns indicative of origin or spread of M. caprae. Altogether, our data confirm M. caprae as a distinct phylogenetic lineage within the Mycobacterium tuberculosis complex.

Supplemental material for this article may be found at http://jcm.asm.org/.

These authors contributed equally to this work.