Application of molecular biological techniques for detection of epizootic hemorrhagic disease virus (EHDV-318) recovered from a sentinel calf in central Sudan.
Epizootic hemorrhagic disease virus (EHDV), isolate 318 (EHDV-318), an untyped virus recovered from a sentinel calf herd at the Khartoum University farm in central Sudan, was characterized using molecular biological techniques. With dot blot hybridization technique, a cDNA probe derived from genome segment 6 of EHDV-2 (Alberta strain) hybridized with RNA from EHDV-318. Application of serogroup-specific EHDV polymerase chain reaction (PCR) to EHDV-318 RNA resulted in specific amplification of a 387 bp PCR product. Amplification product was visualized on ethidium bromide-stained agarose gel. Specificity of the PCR products was confirmed by chemiluminescent hybridization with a non-radiolabelled internal probe. No amplification product or hybridization signal was detected when the serotype-specific EHDV-1 or EHDV-2 PCR-based assays were applied to RNA from EHDV-318. The scientific data presented in this study indicated that cDNA probes and serogroup-specific PCR-based assay can be used to classify the virus as a member of EHDV serogroup, and as serotypically distinct from EHDV-1 and EHDV-2.
Mohammed
M E
ME
Department of Preventive Medicine and Veterinary Public Health, Faculty of Veterinary Science, University of Khartoum, Sudan.
Aradaib
I E
IE
Mukhtar
M M
MM
Ghalib
H W
HW
Riemann
H P
HP
Oyejide
A
A
Osburn
B I
BI
D000818:Animals; D002417:Cattle; D002418:Cattle Diseases / Q000175:diagnosis / Q000453:epidemiology / Q000821:virology*; D015342:DNA Probes; D018164:Hemorrhagic Disease Virus, Epizootic / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D009693:Nucleic Acid Hybridization; D016133:Polymerase Chain Reaction / Q000662:veterinary; D012330:RNA, Double-Stranded / Q000032:analysis; D012367:RNA, Viral / Q000032:analysis*; D012088:Reoviridae Infections / Q000175:diagnosis / Q000453:epidemiology / Q000662:veterinary* / Q000821:virology; D015203:Reproducibility of Results; D013045:Species Specificity; D013397:Sudan / Q000453:epidemiology
D015342:DNA Probes; D012330:RNA, Double-Stranded; D012367:RNA, Viral