Sensitive, semi-nested RT-PCR amplification of fusion gene sequences for the rapid detection and differentiation of Newcastle disease virus.
A rapid, sensitive and specific semi-nested RT-PCR was developed to detect and differentiate virulent and avirulent strains of Newcastle disease virus (NDV). For a total of 67 NDV strains, the results obtained from the semi-nested RT-PCR were consistent with those from nucleotide sequence analysis, plaque forming assays, mean death time (MDT) measurements and intracerebral pathogenicity index (ICPI). Furthermore, 13 class I NDV strains can be characterized by the semi-nested RT-PCR approach, which was feasible by the conventional methods. The detection limit for the semi-nested RT-PCR was two plaque forming units (PFU) both for NDV strain F48E9 in allantoic fluid and for isolate APMV1/ch/ChinaND4031 in oral or cloacal swabs. In conclusion, this semi-nested RT-PCR method offers a new assay for the rapid detection and differentiation of NDVs.
Zhang
Lei
L
Jiangsu Key Laboratory of Zoonosis, College of Veterinary Medicine, Yangzhou University, Yangzhou 225009, PR China.
Pan
Zhiming
Z
Geng
Shizhong
S
Chen
Xiang
X
Hu
Shunlin
S
Liu
Huimo
H
Wu
Yantao
Y
Jiao
Xinan
X
Liu
Xiufan
X
D000818:Animals; D002642:Chick Embryo; D002645:Chickens; D009521:Newcastle Disease / Q000175:diagnosis* / Q000821:virology; D009522:Newcastle disease virus / Q000145:classification* / Q000302:isolation & purification*; D010802:Phylogeny; D012367:RNA, Viral / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods / Q000662:veterinary*; D012680:Sensitivity and Specificity; D014765:Viral Vaccines / Q000276:immunology
D012367:RNA, Viral; D014765:Viral Vaccines