title: Serogrouping and topotyping of Sudanese and United States strains of epizootic hemorrhagic disease virus using PCR.
abstract: The potential use of the recently reported polymerase chain reaction (PCR) protocol for detection of United States epizootic hemorrhagic disease virus (EHDV) serotype 1 (EHDV-1) and serotype 2 (EHDV-2) ribonucleic acid in cell culture and clinical specimens was evaluated for detection of Sudanese EHDV strains. EHDV serotype 5 (EHDV-5) and EHDV, isolate 318 (untyped) designated (EHDV-318), recovered from sentinel calves at the Khartoum University farm (Sudan) were studied. RNA from EHDV-5 and EHDV-318 and a number of EHDV field isolates, propagated in cell cultures, were detected by the described PCR-based assay. The specific 387 bp PCR products were visualized on ethidium-bromide stained agarose gel. Specificity of the PCR products was confirmed by chemiluminescent hybridization with non-radiolabeled internal probe. Amplification product was not detected when the PCR-based assay was applied to RNA from blutongue virus (BTV) prototypes serotypes 2, 10, 11, 13, 16 and 17; total nucleic acid extracts from uninfected BHK-21 cells. The results of this study indicated that the previously described EHDV-PCR assay could be applied for detection of Sudanese as well as United States strains of EHDV serogroup. In addition, the described EHDV-PCR assay could be used as a supportive diagnostic assay to the current conventional virus isolation procedures used for detection of EHDV infection in susceptible ruminants.
authors.affiliation: Department of Veterinary Pathology, Microbiology and Immunology, School of Veterinary Medicine, University of California, Davis 95616, U.S.A.
authors.forename: I E
authors.initials: IE
authors.lastname: Aradaib
authors.forename: M E
authors.initials: ME
authors.lastname: Mohammed
authors.forename: M M
authors.initials: MM
authors.lastname: Mukhtar
authors.forename: H W
authors.initials: HW
authors.lastname: Ghalib
authors.forename: B I
authors.initials: BI
authors.lastname: Osburn
mesh_terms: D000818:Animals; D002460:Cell Line; D003670:Deer / Q000821:virology*; D018164:Hemorrhagic Disease Virus, Epizootic / Q000145:classification* / Q000235:genetics* / Q000302:isolation & purification; D016133:Polymerase Chain Reaction / Q000379:methods* / Q000706:statistics & numerical data; D012330:RNA, Double-Stranded / Q000235:genetics / Q000302:isolation & purification; D012367:RNA, Viral / Q000235:genetics / Q000302:isolation & purification; D012088:Reoviridae Infections / Q000175:diagnosis / Q000662:veterinary* / Q000821:virology; D012680:Sensitivity and Specificity; D013397:Sudan; D014481:United States; D014776:Virus Cultivation
chemical_list: D012330:RNA, Double-Stranded; D012367:RNA, Viral