title: A liquid-hybridization method for typing the Vp4 and Vp7 genes of bovine rotaviruses.
abstract: A simple liquid-hybridization assay was developed which allows assessment of the degree of hybridization between the two serotype-determining genes of the bovine rotavirus strain UK and the homologous genes of the isolate under test. 32P-labelled transcription probes were produced from cloned complementary DNA (cDNA) copies of UK gene segments 4 and 8 and hybridized to double stranded RNA (dsRNA) extracted from rotavirus-positive field samples. Subsequent treatment with ribonuclease A (RNase A), separation of the RNase A-resistant hybrid fragments by polyacrylamide gel electrophoresis (PAGE) and autoradiography yielded a specific, reproducible banding pattern for each isolate. A total of 74 field samples was tested by both the hybridization assay and by an enzyme-linked immunosorbent assay (ELISA) using serotype-specific monoclonal antibodies (Mabs). The results obtained were in excellent agreement and confirmed that serotype G6 rotaviruses predominated. Hybridization of these G6 viruses with the gene 4 probe suggested that viruses with Vp4s related to that of UK rotavirus are also common. The hybridization assay was more sensitive than the ELISA.
authors.affiliation: Moredun Research Institute, Edinburgh, UK.
authors.forename: D L
authors.initials: DL
authors.lastname: Redmond
authors.forename: N F
authors.initials: NF
authors.lastname: Inglis
authors.forename: T A
authors.initials: TA
authors.lastname: Fitzgerald
authors.forename: D R
authors.initials: DR
authors.lastname: Snodgrass
authors.forename: A J
authors.initials: AJ
authors.lastname: Herring
mesh_terms: D000818:Animals; D000956:Antigens, Viral; D002213:Capsid / Q000235:genetics; D036022:Capsid Proteins; D002417:Cattle; D005069:Evaluation Studies as Topic; D005814:Genes, Viral; D009693:Nucleic Acid Hybridization / Q000379:methods*; D015347:RNA Probes; D012367:RNA, Viral / Q000235:genetics; D012401:Rotavirus / Q000145:classification* / Q000235:genetics*; D012703:Serotyping / Q000379:methods; D014773:Virology / Q000379:methods*
chemical_list: D000956:Antigens, Viral; D036022:Capsid Proteins; D015347:RNA Probes; D012367:RNA, Viral; C060339:VP4 protein, Rotavirus; C050779:VP7 protein, Rotavirus