Characterization of monoclonal antibodies directed against the bovine herpesvirus-1 glycoprotein E and use for the differentiation between vaccinatedand infected animals

A panel of seven monoclonal antibodies (MAbs) directed against the bovine h erpesvirus- (BHV-1) glycoprotein E (gE) was obtained. For that purpose, mic e were either tolerized to BHV-1 gE-negative virus and then immunized with wild type BHV-1 or immunized with plasmid DNA expressing the gE and gl glyc oproteins. The MAbs were characterized by their reactivity with the gE prot ein or the gE/gI complex and by competition experiments. Results showed tha t the MAbs were directed against three antigenic domains, two located on th e gE glycoprotein and one on the gE/gI complex. Blocking experiments were p erformed with sera from experimentally vaccinated and infected cattle. A co mpetition was observed between gE-positive bovine sera and six of the seven MAbs. The bovine sera thus recognized two of the three antigenic sites. Fi eld sera were then tested in blocking enzyme-linked immunosorbent assay usi ng one horseradish peroxidase-conjugated MAb. A specificity of 98.2% and a sensitivity of 98.2% compared to the commercially available test were obser ved. (C) 2001 Elsevier Science B.V. All rights reserved.