COMPETITIVE-INHIBITION ENZYME-LINKED-IMMUNOSORBENT-ASSAY FOR ANTIBODYIN SHEEP AND OTHER RUMINANTS TO A CONSERVED EPITOPE OF MALIGNANT CATARRHAL FEVER VIRUS

Malignant catarrhal fever (MCP) is a severe, usually fatal, acute syst emic disease syndrome of certain domestic and wild ruminants caused by members of the family Gammaherpesvirinae. Two distinct but closely re lated viruses cause clinically indistinguishable syndromes: one that i s indigenous to the wildebeest and the other that apparently is indige nous to domestic sheep. Neither the pathogenesis nor the epidemiology of sheep-associated MCF (SA-MCF) is understood, primarily because of a lack of adequate detection methods for the etiologic agent or antibod y against it. No acceptably documented isolates of SA-MCF virus have b een reported, and existing antibody assays suffer from significant cro ss-reactivity with other viruses. As a basis for a specific serologic assay, an attempt was made to identify an epitope conserved among all isolates of MCF viruses, by using a monoclonal antibody (MAb) produced against a previously reported U.S. isolate of MCF virus. A MAb (15-A) which bound a conserved epitope present on all four isolates of MCF v irus examined was found. MAb 15-A did not react with eight common shee p and goat viruses or five common bovine viruses. Immunoprecipitation revealed that the 15-A epitope was located on the viral glycoprotein c omplex, with molecular masses of 115, 110, 105, 78, and 45 kDa. Sera f rom experimentally and naturally infected animals which yielded a simi lar glycoprotein complex immunoprecipitation pattern competed with MAb 15-A for its epitope. A competitive inhibition enzyme-linked immunoso rbent assay (ELISA) based on MAb 15-A was therefore developed. The ass ay detected antibody in inapparently infected sheep and in cattle, dee r, and bison with clinical MCF. Of the 149 serum samples from sheep as sociated with MCF outbreaks, 88 (55%) were seropositive by competitive inhibition ELISA.