Lh. Lee et al., SINGLE-TUBE, NONINTERRUPTED REVERSE TRANSCRIPTION-PCR FOR DETECTION OF INFECTIOUS BURSAL DISEASE VIRUS, Journal of clinical microbiology, 32(5), 1994, pp. 1268-1272
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.