ELISA for bovine herpesvirus 1 (BHV1) antibody with HIRT supernatant

Detailed studies were made of the ability of HIRT supernatant (HS) from bovine herpesvirus 1 (BHV1) infected cultures to sensitize plates for enzyme-linked immunosorbent assay (ELISA). The ultracentrifuged pellet of HS had less sensitizing activity than the supernatant, but the antigen was removed completely by 0.22 micron filters and to some extent by 0.45 micron filters; it was minimally affected by sonication; it was destroyed by the action of Pronase but not by DNAase I when a kallikrein inactivator was added and the mixture incubated; incubation with DNAase II had no effect. Thus the presence of DNA was not required for the sensitizing activity of HS and the antigens recognized by antibodies in HS in ELISA were directed to its protein component. Strong reactions were given in immunoblotting of HS from BHV1 infected tissue cultures with anti-BHV1 glycoprotein monoclonal IgG, but HS from uninfected tissue cultures did not react with the same monoclonal IgG.