Immunohistochemical Staining of Influenza Virus in Tissues
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Immunohistochemical methods are commonly used for studying the pathogenesis of influenza A virus by allowing the identification of sites of replication of the virus in infected tissues and the correlation with the histopathological changes observed. In this chapter, the materials and procedures for performing immunohistochemical detection of influenza virus antigens in tissues using a biotin–streptavidin detection method are provided. The technique involves the following steps: heat-induced antigen retrieval, binding of a primary antibody to the virus antigen, antibody–antigen complex binding by a biotinylated secondary antibody, and binding of an enzyme–streptavidin conjugate. The enzyme is then visualized by application of the substrate chromogen solution to produce a colorimetric reaction with a product that can be visualized. Visualization of influenza virus antigen in tissues is based on chromogen deposition in the nucleus and cytoplasm of infected cells.
Key wordsAvian influenza virus Immunohistochemistry Biotin–streptavidin detection method Influenza
I gratefully acknowledge the assistance of Diane Smith and the personnel from the Food Safety and Inspection Service histopathology laboratory (USDA, Athens, GA) in the development and adaptation of this method.
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