Multiplex RT-PCR for rapid detection and differentiation of class I and class II Newcastle disease viruses
Multiplex RT-PCR for rapid detection and differentiation of class I and class II Newcastle disease viruses
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多重 RT-PCR 用于快速检测和区分 I 类和 II 类新城疫病毒
DOI：
10.1016/j.jviromet.2010.10.017
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发表时间：
2011-01-01
影响因子：
3.1
通讯作者：
Wang, Zhiliang
中科院分区：
文献类型：
--
作者：
Liu, Hualei;Zhao, Yunling;Wang, Zhiliang
建立了鉴别纽卡斯尔病毒（NDV）Ⅰ、Ⅱ类毒株的多重RT-PCR方法。该方法具有较高的特异性和灵敏度。对2009年获得的67株NDV野毒株进行多重RT-PCR检测，结果与核苷酸序列测定和系统发育分析结果一致。根据F基因部分序列构建的系统发育树将67株NDV野毒株分为两类。27个NDV分离物被归为I类，并鉴定出两种基因型。大多数I类分离株被确定为基因型3，但分离株NDV 09 -034除外，其属于基因型2。将40株Ⅱ类NDV分离株分为3种基因型，即基因型VII（27株）、基因型I（2株）和基因型II（11株）。II类中基因型I和II的分离株显示与中国常用的商业疫苗株相关。根据F基因切割位点的序列基序，预测基因型VII的所有分离株都是毒性的。近年来，这种基因型已成为中国大部分ND暴发的主要原因。总之，这种多重RT-PCR提供了一种新的快速检测和区分这两类NDV分离株。(C)2010 Elsevier B. V.保留所有权利。
A multiplex RT-PCR was developed for detection and differentiation of class I and class II strains of Newcastle disease virus (NDV). The method was shown to have high specificity and sensitivity. The results obtained from the multiplex RT-PCR for a total of 67 NDV field isolates obtained in 2009 were consistent with those obtained by nucleotide sequencing and phylogenetic analysis. A phylogenetic tree based on the partial sequences of the F gene revealed that the 67 field isolates of NDV could be divided into two classes. Twenty-seven NDV isolates were grouped into class I, and two genotypes were identified. Most of the class I isolates were determined to be of genotype 3, with the exception of isolate NDV09-034, which belonged to genotype 2. Forty class II NDV isolates were divided into three genotypes, namely genotype VII (27 isolates), genotype I (2 isolates) and genotype II (11 isolates). Isolates of genotypes I and II in class II were shown to be related to commercial vaccine strains used commonly in China. All isolates of genotype VII were predicted to be virulent, on the basis of the sequence motif at the cleavage site of the F gene. This genotype has become predominantly responsible for most outbreaks of ND in China in recent years. In conclusion, this multiplex RT-PCR provides a new assay for rapid detection and differentiation of both classes of NDV isolates. (C) 2010 Elsevier B.V. All rights reserved.