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United States Department of Agriculture

Agricultural Research Service

Title: Diagnosis of Mycobacterium Bovis Infection Using Formalin-Fixed Paraffin Embedded Tissues

Authors
item Miller, Janice
item Rhyan, J - USDA, APHIS, PL, NVSL
item Jenny, A - USDA, APHIS, PL, NVSL
item Henderson, L - USDA, APHIS, VBL, NVSL

Submitted to: International Symposium on Bovine Tuberculosis in Animals and Humans
Publication Type: Proceedings
Publication Acceptance Date: May 11, 1995
Publication Date: N/A

Technical Abstract: Immunohistochemistry (IHC) and polymerase chain reaction (PCR) procedures were developed to detect M. bovis infection in paraffin sections of formalin-fixed tissues or cell cultures. A rabbit antipeptide antibody to MPB70 was used for IHC. Diffuse cytoplasmic staining was observed in many macrophages and giant cells of granulomatous lesions in lymph nodes from M. .bovis-infected cattle. In tests for specificity, some staining was observed in lesions that contained Coccidioides, unknown fungi, or M. paratuberculosis. PCR was performed with primers for a 123 bp segment of IS6110. The test was able to detect M. bovis that were present in large numbers in bovine macrophages infected in vitro and in lymph node from an experimentally infected pig. However, macrophages infected with a small number of bacteria and culture-positive bovine lymph nodes that had few acid-fast organisms gave negative results.

Last Modified: 9/2/2014
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