Application of monoclonal antibody for the immunohistochemical detection of Newcastle disease virus in the chicken Public Deposited

http://ir.library.oregonstate.edu/concern/graduate_thesis_or_dissertations/tx31qn34h

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  • The detection and identification of Newcastle disease virus (NDV) has never been straightforward. Serologically indistinguishable pathotypes produce a wide spectrum of disease that ranges from a severe respiratory, neurologic, or enteric disease with high mortality to an asymptomatic infection. Extensive use of ND vaccine strains further complicates the serological differentiation of field isolates. Recently, monoclonal antibodies (MAbs) have been produced in some laboratories which are capable of detecting antigenically different pathotypes (Russell 1983). Our major objective of this study was to apply immunohistological techniques for rapid detection of NDV antigen using monoclonal or polyclonal antibody in infected chickens. Two methods of immunocytochemistry, immunofluorescence (IF) and alkaline phosphatase-monoclonal anti-alkaline phosphatase (APAAP), were used to demonstrate Newcastle disease virus (NDV) antigen in impression smears and formalin-fixed, paraffin-processed sections of lungs and tracheas obtained from infected chickens. Specific immunofluorescent and alkaline phosphatase staining with a monoclonal antibody was detected within the cytoplasm of epithelial cells of trachea or secondry and tertiary bronchi. The results were compared between the above mentioned techniques and virus isolation. Immunofluorescent staining of impression smears was the easiest approach. Specific staining pattern with MAb was confirmatory in comparison with polyclonal antibodies which generally showed varying degree of nonspecific staining. The staining with APAAP was permanent, and it was possible to correlate histopathological changes with actual location of antigen in situ. These techniques appeared potentially useful for rapid and definitive diagnosis of Newcastle disease in the future.
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