Submitted to: Book Chapter
Publication Type: Book / Chapter
Publication Acceptance Date: 1/15/2007
Publication Date: 1/15/2008
Citation: Pantin Jackwood, M.J. 2008. Immunohistochemical staining of avian influenza viruses in tissues. In: Spackman, E., editor. Avian Influenza Virus. Methods in Molecular Biology. Humana Press, Totowa, NJ. p. 77-83.
Technical Abstract: Immunohistochemical methods are commonly used for studying the pathogenesis of avian influenza 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 methods for performing immunohistochemical detection of avian influenza virus antigens in tissues are outlined. The technique involves the following steps: heat induced antigen retrieval, binding of a primary antibody to a viral specific 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 end product. Demonstration of avian influenza virus antigen in tissues is based on chromogen deposition in the nucleus, which is often accompanied by chromogen deposition within the cytoplasma.