Detection of bluetongue virus using a cDNA probe derived from genome segment 4 of bluetongue virus serotype 2.
The double-stranded (ds) RNA genome segment 4 of bluetongue virus (BTV) serotype 2 was cloned and used as a serogroup-specific complementary (c) DNA probe for BTV diagnosis. A cDNA representing a 60% copy of genome segment 4 BTV-2 prototype was produced. The specificity of the cDNA probe was determined by hybridizing this probe to a northern blot of dsRNA (separated by polyacrylamide gel electrophoresis) of plaque-purified BTV-2 prototype. This cDNA probe was then used to hybridize to the RNA samples. Because the probe hybridized to all BTV samples but not to epizootic hemorrhagic disease virus samples, it appears to be a group-specific probe that could be used in BTV diagnosis.
Chinsangaram
J
J
Department of Veterinary Pathology, School of Veterinary Medicine, University of California, Davis 95616.
Hammami
S
S
Osburn
B I
BI
D000818:Animals; D015152:Blotting, Northern; D001820:Bluetongue virus / Q000235:genetics / Q000302:isolation & purification*; D002460:Cell Line; D003001:Cloning, Molecular; D015342:DNA Probes; D004591:Electrophoresis, Polyacrylamide Gel; D005347:Fibroblasts; D015151:Immunoblotting; D009693:Nucleic Acid Hybridization; D012367:RNA, Viral / Q000032:analysis*; D012680:Sensitivity and Specificity; D014709:Vero Cells
D015342:DNA Probes; D012367:RNA, Viral