| Title |
Application of antibody to recombinant polypeptide a362 and of recombinant DNA probe F57 for diagnosis of Johne's disease. |
| Author(s) |
Buergelt CD,
Mathelart F,
Coetsier C,
Vannuffel P,
Haveau X,
Limbourg B,
Denef JF,
Cocito C.
|
| Institution(s) |
Depart Pathobiol, University of Florida, USA; Depart Histology, Catholic University of Louvain, Brussels, Belgium.
|
| Source |
Fifth International Colloquium on Paratuberculosis
|
| Section |
4:
Diagnostic Approaches to paratuberculosis
|
| Abstract |
The expressed product (a362) of a M. ptbc gene coding for a 34-kDa protein (P34) was used to generate polyclonal antibody in rabbits and monoclonal antibody in rats. The polyclonal antibody was tested for potential to immunohistochemically detect antigen of M. ptbc in intestinal and lymph node tissues of infected cattle. Tissues were subjected to different fixation methods, including cryostat sectioning. For comparison, conventional staining methods were included. The recombinant DNA probe F57 was tested on fresh tissues via PCR and DNA hybridization. Preliminary results obtained indicated that the immunohistochemical detection of M. ptbc antigen was strong in pluribacillary and sporadic in paucibacillary cases. When compared with conventional stains, the immunohistochemical signal was more sensitive in paucibacillary cases. Choice and time of fixation were critical for detection. The DNA probe F57 was specific and sensitive to detect Johne's disease antigen in fresh and paraffin tissues from infected animals.
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