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United States Department of Agriculture

Agricultural Research Service

Title: Comparison of Polyclonal Antibodies to Three Different Preparations of Mycobacterium Paratuberculosis in Immunohistochemical Diagnosis of Johne's Disease in Cattle

Authors
item Stabel, Judith
item Ackermann, Mark
item Goff, Jesse

Submitted to: International Colloquium on Paratuberculosis
Publication Type: Abstract Only
Publication Acceptance Date: October 4, 1996
Publication Date: N/A

Technical Abstract: Polyclonal antisera were raised in rabbits against preparations of live and heat-killed Mycobacterium paratuberculosis as well as cell-wall proteins of M. paratuberculosis and evaluated as diagnostic tools in immunohistochemical staining of bovine tissue. Live preparations of M. paratuberculosis (LMp) were inoculated at 10*9/ml either intraperitoneal or rintravenous. Heat-killed M. paratuberculosis (HKMp) was prepared by treatment of bacteria at 85C for 10 minutes. Cell-wall proteins were isolated from M. paratuberculosis and conjugated to keyhole limpet hemocyanin to improve antigenicity (KLH-CWPMp). Heat-killed and KLH-CWPMp preparations were emulsified in incomplete Freund's adjuvant before subcutaneous inoculation of rabbits. At the terminal bleed, antibody titers were higher for HKMp and KLH-CWPMp rabbits compared to the LMp inoculum (1:1024 vs 1:64). The KLH-CWPMp antibody did not cross-react with hM. bovis-infected tissues. Sensitivity and specificity of immunohistochemical detection of paratuberculosis in bovine tissues was much higher for the KLH-CWPMp polyclonal antibody.

Last Modified: 8/29/2014
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