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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

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: 4/19/2015
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