Rapid detection and differentiation of Newcastle disease virus isolates by a triple one-step RT-PCR.
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.
Wang
Z
Z
Applied Biotechnology Division, Onderstepoort Veterinary Institute, South Africa.
Vreede
F T
FT
Mitchell
J O
JO
Viljoen
G J
GJ
D000818:Animals; D001717:Birds; D002642:Chick Embryo; D009521:Newcastle Disease / Q000821:virology*; D009522:Newcastle disease virus / Q000145:classification / Q000235:genetics / Q000302:isolation & purification* / Q000472:pathogenicity; D012367:RNA, Viral / Q000032:analysis; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000662:veterinary; D012680:Sensitivity and Specificity; D013047:Specific Pathogen-Free Organisms; D014774:Virulence
D012367:RNA, Viral