2011—2016年浙江省猪流行性腹泻病毒S基因与ORF3基因遗传进化分析

发布时间：2018-03-03 01:33

本文选题：猪流行性腹泻病毒　切入点：S基因　出处：《浙江农业学报》2017年03期 　论文类型：期刊论文

【摘要】：为调查近年来浙江省猪流行性腹泻病毒流行毒株与疫苗株的S基因和ORF3基因遗传变异情况,2011—2016年,从浙江省各地区采集疑似猪流行性腹泻病料并提取其RNA,用3对特异性引物对S基因和ORF3基因进行RT-PCR扩增、克隆及测序,并对其序列变化、遗传进化情况和抗原位点进行分析。结果表明:与国内疫苗株CV777相比,12个样品株的S基因存在15个核苷酸插入和6个核苷酸缺失,导致氨基酸序列存在5个氨基酸插入(~(58)QGVN~(61)和~(140)N)和2个氨基酸缺失(~(163)DI~(164)),且在主要突变区S1区的2个中和表位(499-638和764-771 aa)有8处氨基酸突变;样品株的ORF3基因有7处氨基酸变异。遗传进化分析结果显示,12株分离株的S基因与亚洲主要疫苗株(国内疫苗株CV777和韩国弱毒疫苗株DR13)的同源性仅为93.3%~94.9%,而与2011—2016年中国流行株(BJ-2011和Hu N)的同源性高达97.8%~99.9%,表明近年来猪流行性腹泻病毒呈现较大程度的变异,可为研制更加有效的疫苗奠定基础。
[Abstract]:In order to investigate the genetic variation of S gene and ORF3 gene of epidemic and vaccine strains of porcine epidemic diarrhea virus in Zhejiang Province in recent years, Suspected porcine epidemic diarrhea materials were collected from Zhejiang Province and their RNAs were extracted. The S and ORF3 genes were amplified by RT-PCR, cloned, sequenced and sequenced with 3 pairs of specific primers. Genetic evolution and antigenic sites were analyzed. The results showed that there were 15 nucleotide insertions and 6 nucleotide deletions in S gene of 12 strains compared with domestic vaccine strains CV777. The results showed that there were 5 amino acid insertions in the amino acid sequence, 1) and 140N) and 2 amino acid deletions, and 8 amino acid mutations were found in the two neutralization epitopes of S1 region (49-638 and 764-771 aa). There were 7 amino acid mutations in the ORF3 gene of the sample strain. The results of genetic evolution analysis showed that the S gene of 12 strains was only 93.3% 94.9 homology with the major Asian vaccine strains (domestic vaccine CV777 and Korean attenuated virus strain DR13), but only 93.3% 94.9 with 2011-2016. The homology of Chinese epidemic strains BJ-2011 and Hu Nis as high as 97.8% and 99.9%, indicating that the swine epidemic diarrhea virus has shown a large degree of variation in recent years. It can lay the foundation for the development of a more effective vaccine.
【作者单位】：甘肃农业大学动物医学院;浙江省农业科学院畜牧兽医研究所;
【基金】：公益技术研究农业项目(2015C32028,2016C32072)
【分类号】：S852.65

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