China Animal Health and Epidemiology Center, Qingdao, Shangdong, China.
J Virol Methods. 2011 Jan;171(1):149-55. doi: 10.1016/j.jviromet.2010.10.017. Epub 2010 Oct 27.
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.
一种用于检测和区分新城疫病毒（NDV）I 型和 II 型毒株的多重 RT-PCR 方法被开发出来。该方法显示出高度的特异性和敏感性。该多重 RT-PCR 方法总共对 2009 年获得的 67 个 NDV 田间分离株进行了检测，其结果与核苷酸测序和系统进化分析的结果一致。基于 F 基因部分序列构建的系统进化树显示，67 个 NDV 田间分离株可分为两类。27 个 NDV 分离株归为 I 类，鉴定出两个基因型。大多数 I 类分离株被确定为基因型 3，除了 NDV09-034 分离株属于基因型 2。40 个 II 类 NDV 分离株分为三个基因型，即基因型 VII（27 个分离株）、基因型 I（2 个分离株）和基因型 II（11 个分离株）。II 类的基因型 I 和 II 分离株与中国常用的商业疫苗株有关。根据 F 基因裂解位点的序列特征，预测所有基因型 VII 的分离株均为强毒力。该基因型已成为近年来中国大多数 ND 暴发的主要原因。总之，该多重 RT-PCR 为快速检测和区分两类 NDV 分离株提供了一种新方法。
