An immunohistochemical staining technique for the detection of marble spleen disease (MSD) viral antigens and other type II avian adenoviral antigens was developed using a mixture of monoclonal antibodies produced against hemorrhagic enteritis (HE) virus and a commercial streptavidin-biotin peroxidase indicator system. This technique was applied to both frozen and formalin-fixed paraffin-embedded tissue sections. The immunohistochemical staining technique was used on tissues from pheasants with experimental MSD, on tissues from a pheasant with natural MSD, and on tissues from turkeys with natural HE. Staining results were compared with routine hematoxylin-and-eosin (H&E) staining. Additional viral inclusions, not detected with H&E, were found in the liver, lung, bone marrow, and kidney sections using the immunohistochemical technique. The immunohistochemical technique was highly specific and sensitive for the detection of type II adenoviral antigen, and it appears to be useful for studying the pathogenesis of these diseases and for retrospective evaluation of routinely processed diagnostic tissue samples.
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