Fluorescent and electron-microscopy immunoassays employing polyclonal and monoclonal antibodies for detection of goose parvovirus infection

Polyclonal antibodies (PAbs) raised in geese and eight mice hybridomas secr eting monoclonal antibodies (MAbs) against the goose parvovirus (GPV) were prepared. They were used for development of immunofluorescence (IF) and imm unoelectron-microscopic (IEM) techniques to demonstrate the GPV infection i n infected organs and biological fluids. The GPV antigens were established by immunofluorecence within the nuclei and the cytoplasm of many infected c ells of the chorioallantoic membrane of goose and Peckin duck embryos, live r and heart of mortally diseased goslings. By means of IEM it was possible to detect the GPV in native organ homogenate supernatants and allantoic flu ids. All techniques used in the study could be successfully applied for rap id diagnosis of the GPV infection. The test systems on the basis of MAbs sh ould, however, be preferred. By means of immunoblotting (IB) using PAbs and MAbs four viral proteins (VP) with MW 88, 77, 65 and 60 kDa were demonstra ted. Contrary to the others the VP with MW 65 kDa was the most antigenicall y reactive though invisible in the SDS-PAGE and Coomassie-blue dye-stained preparations. (C) 1999 Elsevier Science B.V. All rights reserved.