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贵州省猪流行性腹泻病毒ORF3、M基因克隆及序列分析
引用本文:冯旭芳,周碧君,张双翔,王跃章,文明,程振涛,毛黎红,王开功. 贵州省猪流行性腹泻病毒ORF3、M基因克隆及序列分析[J]. 中国畜牧兽医, 2016, 43(2): 356-362. DOI: 10.16431/j.cnki.1671-7236.2016.02.010
作者姓名:冯旭芳  周碧君  张双翔  王跃章  文明  程振涛  毛黎红  王开功
作者单位:1. 贵州大学动物科学学院, 贵阳 550025;2. 贵州省生猪质量安全工程技术研究中心, 贵阳 550025;3. 贵州省动物疫病预 防控制中心, 贵阳 550001;4. 贵州都匀市黔昌畜牧发展有限责任公司, 都匀 558000;5. 花溪区农业局, 贵阳 550025
基金项目:贵州省生猪质量安全工程技术研究中心建设项目"生猪健康养殖实验室建设与技术研发"(黔科合农C字[2011]4022号);贵州省科学技术厅农业攻关项目"贵州省仔猪腹泻性疫病风险分析及综合防控技术研究"(黔科合NY[2015]3009-1号);"贵州省动物疫病防控与兽医公共卫生保障科技创新人才团队"(黔科合人才团队[2015]4016号);贵州省动物疫病与兽医公共卫生重点实验室培育(黔科合计Z字[2011]4008号);贵州省农业委员会科技计划项"贵州省2015年重大动物疫病省级定点监测"(2015-01号)
摘    要:为了解贵州省猪流行性腹泻病毒(porcine epidemic diarrhea virus,PEDV)毒株ORF3及M基因的遗传变异情况,试验于2014年4月-2015年3月从贵州省5个地区采集105份腹泻仔猪的粪便,应用RT-PCR方法进行PEDV检测,从中选择8份PEDV阳性样本,扩增其ORF3及M基因,测序并进行序列比对分析.结果显示,从采集的105份粪便样本中可检出75份PEDV阳性样本,阳性率为71.43%;8株PEDV贵州株ORF3及M基因序列均无碱基缺失或插入;ORF3基因核苷酸及推导的氨基酸同源性在95.1%~100.0%与95.1%~99.6%之间,M基因核苷酸及推导的氨基酸同源性在98.4%~100.0%与98.7%~100.0%之间;氨基酸系统进化树分析结果显示,2014~2015年贵州流行株与近年来中国毒株、韩国毒株及泰国毒株亲缘关系较近,与疫苗株Attenuated DR13及CV777株亲缘关系较远.提示目前贵州省仔猪腹泻病原主要是PEDV,且为PEDV强毒株.

关 键 词:猪流行性腹泻病毒  ORF3基因  M基因  克隆  序列分析  
收稿时间:2015-07-28

Cloning and Sequence Analysis of ORF3 and M Gene of Porcine Epidemic Diarrhea Virus in Guizhou Province
FENG Xu-fang,ZHOU Bi-jun,ZHANG Shuang-xiang,WANG Yue-zhang,WEN Ming,CHENG Zhen-tao,MAO Li-hong,WANG Kai-gong. Cloning and Sequence Analysis of ORF3 and M Gene of Porcine Epidemic Diarrhea Virus in Guizhou Province[J]. China Animal Husbandry & Veterinary Medicine, 2016, 43(2): 356-362. DOI: 10.16431/j.cnki.1671-7236.2016.02.010
Authors:FENG Xu-fang  ZHOU Bi-jun  ZHANG Shuang-xiang  WANG Yue-zhang  WEN Ming  CHENG Zhen-tao  MAO Li-hong  WANG Kai-gong
Affiliation:1. College of Animal Science, Guizhou University, Guiyang 550025, China;2. Guizhou Pig Quality Safety Engineering Research Center, Guiyang 550025, China;3. Animal Epidemic Disease Control Center of Guizhou Province, Guiyang 550001, China;4. Qian Chang Stockbreeding Develop Limited Liability Company of Duyun, Duyun 558000, China;5. Agricultural Bureau of Huaxi, Guiyang 550025, China
Abstract:To study the genetic variations of porcine epidemic diarrhea virus (PEDV) ORF3 and M gene in Guizhou province,we used RT-PCR method to detect PEDV in the dung what collected from diarheal porket in five regions of Guizhou province between April 2014 to March 2015,then selected eight positive samples,cloned and sequenced their ORF3 and M gene.The results showed that 75 samples were positive for PEDV,and the positive rate was 71.43%.The result of sequencing showed that ORF3 and M gene were intact;ORF3 gene shared from 95.1% to 100.0% nucleotide identity and 95.1% to 99.6% amino acid identity,and M gene shared from 98.4% to 100.0% nucleotide identity and 98.7% to 100.0% amino acid identity with eight PEDV Guizhou strains.Phylogenetic analysis revealed that Guizhou strains seem to be closely related to Chinese strains,Korean strains and Thai strains,and there were genetically different from the vaccine strains attenuated DR13 and CV777.The results suggested that in rencent years the mainly etiology of orket diarrhea was velogenic PEDV.
Keywords:porcine epidemic diarrhea virus  ORF3 gene  M gene  cloning  sequence analysis  
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