title: Diagnostic complementary DNA probes for genome segments 2 and 3 of epizootic hemorrhagic disease virus serotype 1.
abstract: Potential diagnostic complementary DNA (cDNA) clones of gene segments 2 and 3 from epizootic hemorrhagic disease virus serotype 1 (EHDV-1) have been produced. Individual segments of EHDV-1 were isolated, denatured with methylmercury hydroxide, and polyadenylated. The polyadenylated RNA was reverse-transcribed and self-hybridized into duplex structures, and the incomplete ends were repaired. The resulting product was then cloned into the plasmid vector pBR322, using the complementary tailing method. Two clones, 1 from segment 2 (E1-2-10) and 1 from segment 3 (E1-3-16) were isolated, colony-purified, and characterized by cDNA/RNA blot hybridization and endonuclease restriction analysis. The cDNA clones of RNA segment 3 of EHDV-1 cross hybridized with the corresponding segment of EHDV serotype 2 by results of cDNA/RNA blot hybridization, but not with RNA of bluetongue virus serotypes isolated in the United States. After cDNA/RNA dot-blot hybridization analysis of 17 EHDV field strains, the segment-2 clone was found to be serotype-specific, whereas the segment-3 clone was serogroup-specific.
authors.affiliation: US Department of Agriculture, Agricultural Research Service, Laramie, WY 82071-3965.
authors.forename: W C
authors.initials: WC
authors.lastname: Wilson
authors.forename: A
authors.initials: A
authors.lastname: Fukusho
authors.forename: P
authors.initials: P
authors.lastname: Roy
mesh_terms: D000818:Animals; D001820:Bluetongue virus / Q000145:classification; D015342:DNA Probes / Q000138:chemical synthesis / Q000235:genetics*; D004279:DNA, Viral / Q000235:genetics*; D004587:Electrophoresis, Agar Gel; D004926:Escherichia coli / Q000235:genetics; D015698:Genomic Library; D006476:Orthohantavirus / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification; D015151:Immunoblotting; D009693:Nucleic Acid Hybridization; D012330:RNA, Double-Stranded / Q000032:analysis / Q000235:genetics* / Q000302:isolation & purification; D012367:RNA, Viral / Q000235:genetics*
chemical_list: D015342:DNA Probes; D004279:DNA, Viral; D012330:RNA, Double-Stranded; D012367:RNA, Viral