Rapid detection and differentiation of Newcastle disease virus isolates by a triple one-step RT-PCR
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Date
Authors
Wang, Zheng-Wei
Vreede, F.T.
Mitchell, J.O.
Viljoen, G.J.
Journal Title
Journal ISSN
Volume Title
Publisher
Published jointly by the Agricultural Research Council, ARC-Onderstepoort Veterinary Institute and the Faculty of Veterinary Science, University of Pretoria.
Abstract
A triple one-step RT-PCR was developed to screen and differentiate virulent from avirulent Newcastle disease virus (NDV) isolates. Three sets of oligonucleotides were designed, each specific for amplifying NDV fusion protein gene-specific RNA from virulent, avirulent or all isolates respectively. The sensitivity of one-step RT-PCR was determined using viral RNA extracted from serially diluted NDV-infected allantoic fluid and found to be 10ˉ⁵ HA units. Application of one-step RT-PCR to various NDV samples, including wild-type virulent isolates and avirulent vaccine strains, demonstrated the potential for rapid identification (3-4 h) of NDV isolates as well as the differentiation of virulent from avirulent strains.
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Keywords
Veterinary medicine, Detection, Identification, Newcastle disease virus (NDV), RT-PCR, Triton X-100 differentiation of NDV isolates
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Citation
Wang, Z, Vreede, FT, Mitchell, JO & Viljoen, GJ 2001, 'Rapid detection and differentiation of Newcastle disease virus isolates by a triple one-step RT-PCR’. Onderstepoort Journal of Veterinary Research, vol. 68, no. 2, pp. 131-134.