Title: Immunohistochemical staining of avian influenza virus in tissues Author
Submitted to: Animal Influenza Virus
Publication Type: Book / Chapter
Publication Acceptance Date: November 18, 2013
Publication Date: June 12, 2014
Citation: Pantin Jackwood, M.J. 2014. Immunohistochemical staining of avian influenza virus in tissues. In: Spackman, E, editor. Animal Influenza Virus: Methods in Molecular Biology. Volume 1161. London: Humana Press. p. 51-58. Interpretive Summary: Viral antigens in tissues of infected animals can be detected by the use of specific antibodies labeled with dyes or enzymes. This detection process is known as immunohistochemistry (IHC). This antigen detection system permits to correlate virus replication with the corresponding pathologic cellular changes. For this reason, IHC is frequently used in avian influenza (AI) virus pathogenesis studies and has allowed for greater understanding of the pathogenic mechanisms involved during AI virus infection by the detection of viral antigens in target tissues. In this chapter, we provide the materials and methods needed for performing immunohistochemical detection of AI virus antigens in formalin fixed-paraffin embedded tissues.
Technical Abstract: Immunohistochemical methods are commonly used for studying the pathogenesis of avian influenza (AI) 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 AI virus antigens in tissues 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 end product. Demonstration of AI virus antigen in tissues is based on chromogen deposition in the nucleus and cytoplasm of infected cells.