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Clinical and Diagnostic Laboratory Immunology, July 1998, p. 446-451, Vol. 5, No. 4
1071-412X/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Detection of Mycobacterium avium subsp. paratuberculosis in Infected Tissues by New Species-Specific Immunohistological Procedures

Christophe Coetsier,1 Xavier Havaux,2 Francois Mattelard,1 Sanaa Sadatte,1 Francoise Cormont,2 Klaus Buergelt,3 Bernard Limbourg,4 Dominique Latinne,2 Herve Bazin,2 Jean-Francois Denef,1 and Carlo Cocito1,*

Histology Unit1 and Experimental Immunology Unit,2 University of Louvain Medical School, Brussels, and Centre de Dépistage des Maladies du Bétail, Erpent,4 Belgium, and College of Veterinary Medicine, University of Florida, Gainesville, Florida3

Received 1 October 1997/Returned for modification 25 November 1997/Accepted 6 April 1998

We have previously described the cloning and sequencing of a gene portion coding for the terminal part of a 34-kDa protein of Mycobacterium avium subsp. paratuberculosis, the etiological agent of Johne's disease (P. Gilot, M. De Kesel, L. Machtelinckx, M. Coene, and C. Cocito, J. Bacteriol. 175:4930-4935, 1993). The recombinant polypeptide (a362) carries species-specific B-cell epitopes which do not cross-react with other mycobacterial pathogens (M. De Kesel, P. Gilot, M.-C. Misonne, M. Coene, and C. Cocito, J. Clin. Microbiol. 31:947-954, 1993). The present work describes the preparation of polyclonal and monoclonal antibodies directed against a362 and the use of these immunoglobulins for histopathological diagnosis of Johne's disease. The new immunohistological procedures herewith detailed proved to be able to identify M. avium subsp. paratuberculosis antigens in the intestinal tissues and lymph nodes of cattle affected by either the paucibacillary or pluribacillary form of the disease. They yielded negative responses not only with healthy animals but also with those affected by tuberculosis (Mycobacterium bovis). Both immunohistological procedures proved to be as sensitive as or more sensitive than Ziehl-Neelsen staining and, in addition, to be endowed with species specificity.

* Corresponding author. Mailing address: GEMO-ISTO-UCL-5225, Ave. Mounier 52, Brussels 1200, Belgium. Phone and fax: 32-2-764-5225.

Clinical and Diagnostic Laboratory Immunology, July 1998, p. 446-451, Vol. 5, No. 4
1071-412X/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.


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