中国畜牧兽医

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2012年河南省猪流行性腹泻病毒分离株S1基因遗传变异分析

陈慧娟1,吴志明2,闫若潜2,赵雪丽2,赵明军2,周兵强2,孔刚锐2,闫志玲2,程俊贞2,靳利粉1   

  1. (1.河南农业大学牧医工程学院,河南郑州 450002;2.河南省动物疫病预防控制中心,河南郑州 450008)
  • 收稿日期:2013-08-16 出版日期:2014-03-20 发布日期:2014-05-15
  • 通讯作者: 吴志明,研究员。E-mail:wuzhiming6@sina.com
  • 作者简介:陈慧娟(1986—),女,河南人,硕士生,研究方向:动物疫病分子生物学。
  • 基金资助:

    河南省科技攻关计划项目“猪流行性腹泻病毒分子流行病学调查及其致病性研究”(132102110159)。

Genetic Variation Analysis of S1 Genes of Porcine Epidemic Diarrhea Virus Isolated from Henan in 2012

CHEN Hui-juan1, WU Zhi-ming2, YAN Ruo-qian2, ZHAO Xue-li2, ZHAO Ming-jun2, ZHOU Bing-qiang2, KONG Gang-rui2, YAN Zhi-ling2, CHENG Jun-zhen2, JIN Li-fen1   

  1.  (1.College of Husbandry and Veterinary, Henan Agricultural University, Zhengzhou 450002, China;2.Henan Center for Animal Disease Control and Prevention, Zhengzhou 450008, China)
  • Received:2013-08-16 Online:2014-03-20 Published:2014-05-15

摘要: 为分析2012年河南省猪流行性腹泻病毒(PEDV)的遗传变异情况,本研究设计合成2对扩增S1基因的特异性引物,利用RT-PCR方法,对2012年分离的5株PEDV毒株的S1基因进行扩增、克隆和序列测定,并对其进行遗传进化分析。结果表明,5株PEDV S1基因核苷酸和氨基酸同源性分别为98.3%~99.5% 和97.1%~98.9%;与2011年以来国内登录的PEDV流行变异株核苷酸同源性为88.6%~98.0%,氨基酸同源性为85.3%~98.7%;与经典毒株CV777核苷酸同源性为88.7%~89.1%,氨基酸同源性为87.3%~88.4%。5株分离株S1基因均存在相同的插入和缺失,与2011年以来国内登录的流行变异毒株相比没有大的变异,但与参考毒株CV777相比,在163—166 bp处有3个核苷酸插入;在173—174 bp之间有9个核苷酸插入;在405—406 bp之间有3个核苷酸插入;在463—464 bp之间有3个核苷酸缺失,这些位点核苷酸的插入和缺失导致了其编码的氨基酸相应改变。S1基因系统进化树分析结果表明,5株分离株与2011年以来国内登录的流行变异毒株均属于基因Ⅲ群,与2011年国内登录的少部分PEDV基因Ⅰ群流行毒株的核苷酸同源性为88.6%~89.3%,氨基酸同源性为85.3%~86.9%,而经典毒株CV777所在分支群属于基因Ⅱ群。本试验结果表明,2011年以来,中国流行的PEDV毒株中同时有基因Ⅲ和Ⅰ群的存在,但以基因Ⅲ群毒株为主,其致病性和抗原性差异仍有待进一步研究。

关键词: 猪流行性腹泻病毒; S1基因; 遗传变异; 分析

Abstract: We designed and synthesized two pairs of specific primers amplified S1 gene by RT-PCR method to investigate the variation in S1 genes of porcine epidemic diarrhea virus (PEDV) in 2012 in Henan.S1 genes of 5 PEDV strains were cloned and sequenced, and their phylogenetic trees were analyzed from different swine breeding farms. Sequences analysis showed that S1 genes shared 98.3% to 99.5% nucleotide identities and 97.1% to 98.9% amino acids homologies among five PEDV isolates. Compared with domestic landing PEDV from 2011, the nucleotide homologies were 88.6% to 98.0% and amino acid homologies were 85.3% to 98.7%. Compared with CV777, the nucleotide homologies were 88.7% to 89.1% and amino acid homologies were 87.3% to 88.4%. Homology analysis showed that S1 genes of 5 isolates shared the same genetic mutation, there were the same insertions and deletions. Compared with domestic mutant strains landed from 2011, there was no tendency. However, compared with CV777, there were three nucleotide insertions from 163 to 166 bp, nine nucleotide insertions from 173 to 174 bp, three nucleotide insertions from 405 to 406 bp and three nucleotide deletions from 463 to 464 bp. These insertions and deletions of nucleotides led to its corresponding changes in encoding amino acids. Phylogenetic analysis showed that S1 genes of 5 PEDV strains belonged to the third group. However compared with part of PEDV first group domestic mutant strains landed in 2011, the nucleotide homologies were 88.6% to 89.3% and amino acid homologies were 85.3% to 86.9%. CV777 was the second group. The results showed that S1 genes of PEDV prevalent strain exist in first and third groups, but mainly prevailing third group strains, the pathogenic and antigenic differences of these strains should be further studied.

Key words: porcine epidemic diarrhea virus; S1 gene; genetic variation; analysis