ISOLATION AND CHARACTERIZATION OF A CORONAVIRUS FROM ELK CALVES WITH DIARRHEA

This is the first report of the isolation of a coronavirus from elk ca lves, Two fecal samples from elk calves with diarrhea were shown to be positive for coronavirus-like particles by electron microscopy, and t he particles were propagated in the human rectal tumor-18 cell line, A fter 24 h, syncytia were observed, and cell culture supernatants from both samples showed hemagglutinating activity with mouse erythrocytes. Cells infected with both elk coronavirus (ECV) isolates reacted with Z3A5, a monoclonal antibody against the spike protein of bovine corona virus (BCV), on an indirect fluorescent antibody test, The protein pro files of both ECV isolates were similar to that of BCV as determined b y sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis, On Northern blot analysis, the transcriptional pattern of ECV was typi cal of coronaviruses, with a nested set of transcripts with common 3' end sequences, Based on a published nucleoprotein gene sequence for BC V (Mebus isolate), we arbitrarily designed two primers for amplificati on by PCR, After cloning, the nucleoprotein was sequenced and a high d egree of homology (99%) between the nucleoprotein gene sequences of EC V and BCV was observed, Thus, ECV is closely related genetically and a ntigenically to BCV and will be a new member of antigenic group 2 of t he mammalian coronaviruses, which possess hemagglutinin-esterase prote in.