Single-tube, noninterrupted reverse transcription-PCR for detection of infectious bursal disease virus
Year: 1994
Type: article
Abstract: An assay protocol based on single-tube, noninterrupted reverse transcription-PCR (RT-PCR) for the detection of infectious bursal disease virus (IBDV) is described. After the conditions for RT-PCR had been optimized, a primer set framing a region within the gene coding for IBDV VP2 protein was used to amplify a 318-bp fragment of the IBDV genome. Amplified product was detected with three strains of IBDV, whereas none was obtained from uninfected bursal tissue or seven unrelated avian viruses. 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 ethidium bromide-stained gel. In addition, this assay system was used to detect IBDV in bursal-tissue specimens from commercially reared chickens. The identity of the amplified products from the tissue specimen preparation was determined by using a rapid, simple procedure in which internally nested, end-labeled probes were used.
Source: Journal of Clinical Microbiology
Authors: L H Lee, L J Ting, J H Shien, H K Shieh
Institution: National Chung Hsing University
Language: English
Subfield: Animal Science and Zoology
Domain: Life Sciences
Open Access status: green
OpenAlex ID: W2145845189