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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.


Journal article


Onderstepoort J Vet Res

Publication Date





131 - 134


Animals, Birds, Chick Embryo, Newcastle Disease, Newcastle disease virus, RNA, Viral, Reverse Transcriptase Polymerase Chain Reaction, Sensitivity and Specificity, Specific Pathogen-Free Organisms, Virulence