Competitive inhibition enzyme-linked immunosorbent assay for antibody in sheep and other ruminants to a conserved epitope of malignant catarrhal fever virus

Abstract

Malignant catarrhal fever (MCF) is a severe, usually fatal, acute systemic disease syndrome of certain domestic and wild ruminants caused by members of the family Gammaherpesvirinae. Two distinct but closely related viruses cause clinically indistinguishable syndromes: one that is indigenous to the widebeest and the other that apparently is indigenous 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 antibody against it. No acceptably documented isolates of SA-MCF virus have been reported, and existing antibody assays suffer from significant cross-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 virus examined was found. MAb 15-A did not react with eight common sheep and goat viruses or five common bovine viruses. Immunoprecipitation revealed that the 15-A epitope was located on the viral glycoprotein complex, with molecular masses of 115, 110, 105, 78, and 45 kDa. Sera from experimentally and naturally infected animals which yielded a similar glycoprotein complex immunoprecipitation pattern competed with MAb 15-A for its epitope. A competitive inhibition enzyme-linked immunosorbent assay (ELISA) based on MAb 15-A was therefore developed. The assay detected antibody in inapparently infected sheep and in cattle, deer, and bison with clinical MCF. Of the 149 serum samples from sheep associated with MCF outbreaks, 88 (55%) were seropositive by competitive inhibition ELISA.