Development and characterization of nucleic acid probes to infectious bursal disease viruses

Radiolabeled cDNA probes were prepared using both segments of double-stranded genomic infectious bursal disease virus (IBDV) RNA as template. The probes were synthesized and labeled with 32P using random primers and reverse transcriptase. Probes were prepared to the genomic RNA extracted from a pathogenic serotype 1 virus (ST-C) and from an attenuated serotype 1 vaccine virus (D-78) which is commercially available. These probes were determined to range in size from approximately 200 to 1500 nucleotides in length using a 6% denaturing polyacrylamide gel. The probes were used in a dot hybridization assay and detected approximately 10 ng of IBDV RNA. In addition, they detected genomic RNA from five different subtypes of IBDV serotype 1 and from two serotype 2 viruses. The probes appeared to be specific for viral RNA since hybridization to cell culture nucleic acid was not detected.