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,...

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Bibliographic Details
Published in:Research in veterinary science 2010-10, Vol.89 (2), p.282-289
Main Authors: Zhang, Lei, Pan, Zhiming, Geng, Shizhong, Chen, Xiang, Hu, Shunlin, Liu, Huimo, Wu, Yantao, Jiao, Xinan, Liu, Xiufan
Format: Article
Language:English
Subjects:
Animals
Chick Embryo
Chickens
Disease
disease diagnosis
Fusion protein
Genes
Microbiology
molecular sequence data
Newcastle disease
Newcastle Disease - diagnosis
Newcastle Disease - virology
Newcastle disease virus
Newcastle disease virus - classification
Newcastle disease virus - isolation & purification
nucleotide sequences
pathogen identification
Phylogeny
Poultry
reverse transcriptase polymerase chain reaction
Reverse Transcriptase Polymerase Chain Reaction - methods
Reverse Transcriptase Polymerase Chain Reaction - veterinary
RNA, Viral - genetics
Semi-nested RT-PCR
Sensitivity and Specificity
strain differences
Veterinary medicine
viral fusion proteins
Viral Vaccines - immunology
Viruses
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Summary: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.
ISSN:0034-5288
1532-2661
DOI:10.1016/j.rvsc.2010.02.007