The use of biotin-labeled cDNA probes for the detection of infectious bursal disease viruses.
A cDNA library was prepared from the double-stranded RNA genome of the infectious bursal disease virus (IBDV) strain ST-C. The cDNA molecules were annealed into the plasmid pUC9 and used to transform Escherichia coli strain JM107. A cDNA clone that contained IBDV-specific nucleotide sequences was selected and designated STC-1. Radiolabeled probes were prepared from STC-1 and hybridized to genome segment A of ST-C in a northern blot hybridization assay. The STC-1 cDNA was 448 base pairs in length, and its nucleotide sequence indicated that it is located near the VP-2/VP-4 junction in IBDV genome segment A. Biotin-labeled probes were prepared from STC-1 and used in a dot-blot hybridization assay to detect IBDV. Under relatively low stringency conditions of hybridization, the biotinylated probes detected four subtypes of IBDV serotype 1 and a serotype 2 IBDV isolate.
Jackwood
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DJ
Food Animal Health Research Program, Ohio Agricultural Research and Development Center, Ohio State University, Wooster 44691.
Kibenge
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FS
Mercado
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CC
D000818:Animals; D015152:Blotting, Northern; D002645:Chickens; D003001:Cloning, Molecular; D015342:DNA Probes; D004279:DNA, Viral / Q000032:analysis*; D007243:Infectious bursal disease virus / Q000235:genetics / Q000302:isolation & purification*; D009693:Nucleic Acid Hybridization; D012330:RNA, Double-Stranded / Q000235:genetics; D012367:RNA, Viral / Q000235:genetics; D012087:Reoviridae / Q000302:isolation & purification*
D015342:DNA Probes; D004279:DNA, Viral; D012330:RNA, Double-Stranded; D012367:RNA, Viral