SINGLE-TUBE, NONINTERRUPTED REVERSE TRANSCRIPTION-PCR FOR DETECTION OF INFECTIOUS BURSAL DISEASE VIRUS

An assay protocol based on single-tube, noninterrupted reverse transcr iption-PCR (RT-PCR) for the detection of infectious bursal disease vir us (IBDV) is described. lifter the conditions for RT-PCR had been opti mized, a primer set framing a region within the gene coding for IBDV V P2 protein,cas used to amplify a 318-bp fragment of the IBDV genome. A mplified product was detected with three strains of IBDV, whereas none was obtained from uninfected bursal tissue or seven unrelated avian v iruses. The sensitivity of this RT-PCR was tested with purified viral RNA from three strains of IBDV. The detection limit was 10 fg in an et hidium bromide-stained gel. In addition, this assay system was used to detect IBDV in bursal-tissue specimens from commercially reared chick ens. The identity of the amplified products from the tissue specimen p reparation was determined by using a rapid, simple procedure in which internally nested, end-labeled probes were used.