Rapid detection and differentiation of Newcastle disease virus isolates by a triple one-step RT-PCR |
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| Authors: | Wang Z Vreede F T Mitchell J O Viljoen G J |
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| Institution: | Applied Biotechnology Division, Onderstepoort Veterinary Institute, South Africa. |
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| 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 10(-5) 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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