Exotic and Emerging Avian Viral Diseases Unit, US National Poultry Research Center, US Department of Agriculture, Agricultural Research Service, Athens, GA, USA.
Methods Mol Biol. 2020;2123:29-36. doi: 10.1007/978-1-0716-0346-8_3.
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.
免疫组织化学方法常用于研究甲型流感病毒的发病机制，通过这种方法可以识别病毒在感染组织中的复制部位，并与观察到的组织病理学变化相关联。在本章中，提供了使用生物素-链霉亲和素检测方法在组织中检测流感病毒抗原的免疫组织化学检测的材料和程序。该技术包括以下步骤：热诱导抗原修复、将针对病毒抗原的一抗与病毒抗原结合、用生物素化的二抗结合抗体-抗原复合物、以及酶-链霉亲和素缀合物的结合。然后通过应用底物显色溶液将酶可视化，产生可可视化的产物的比色反应。组织中流感病毒抗原的可视化是基于在感染细胞的细胞核和细胞质中沉积的显色剂。
