猪流行性腹泻病毒河南株ORF3基因分子流行病学分析

发布时间：2018-04-17 03:33

本文选题：猪流行性腹泻病毒 + ORF基因　；参考：《中国兽医学报》2017年08期

【摘要】：为了解猪流行性腹泻病毒(porcine epidemic diarrhea virus,PEDV)河南株的起源和遗传变异,本试验对2014年8月—2015年5月,来自河南省16个规模猪场仔猪表现为严重腹泻、食欲下降和脱水等临床特征的22份病料样品,利用RT-PCR方法,扩增PEDVORF3基因,回收PCR产物克隆至pMD18-T载体,并进行测序分析。结果显示,22份病料样品均能扩增出PEDVORF3基因,其序列长度均为849bp,包含一个完整阅读框(675bp),编码224个氨基酸残基。22个河南株之间核苷酸同源性为95.9%~100%,与欧洲经典毒株CV777同源性介于97.1%~97.9%,存在8个位点发生相同的氨基酸置换,与Truncated CV777株同源性介于94.8%~95.4%。基因遗传进化树分析表明,PEDV毒株可分为2大群,河南株属于第Ⅱ群,整体独立成支,与河南往年流行株、国内分离株、日本、美国及韩国毒株亲缘关系较近,而与CV777株、Truncated CV777株亲缘关系较远。结果表明PEDV河南株ORF3基因存在变异,为PEDVORF3基因的蛋白功能研究和河南省PED的防控提供技术支持。
[Abstract]:In order to understand the origin and genetic variation of porcine epidemic diarrhea virus (PEV) Henan strain, this experiment showed severe diarrhea in piglets from 16 pig farms in Henan province from August 2014 to May 2015.RT-PCR method was used to amplify the PEDVORF3 gene from 22 samples with clinical characteristics such as loss of appetite and dehydration. The PCR product was recovered and cloned into pMD18-T vector and sequenced.The results showed that PEDVORF3 gene could be amplified from 22 samples.The sequence length was 849 BP, containing a complete reading frame of 675 BP, encoding 224 amino acid residues. The nucleotide homology of 22 Henan strains was 95.9%, and the homology with European classical virus strain CV777 was between 97.1 and 97.9. There were 8 amino acid substitutions at the same sites.The homology with Truncated CV777 strain was 94.8%.Genetic phylogenetic tree analysis showed that the PEDV strains could be divided into two groups. Henan strain belonged to the second group, which was independent and related to the previous epidemic strains in Henan Province, the domestic isolates, Japan, the United States and Korea.The relationship between Truncated CV777 strain and CV777 strain was far from that of Truncated CV777 strain.The results showed that the variation of ORF3 gene in Henan strain of PEDV provided technical support for the study of protein function of PEDVORF3 gene and the prevention and control of PED in Henan province.
【作者单位】：河南农业大学牧医工程学院;吉林大学动物医学学院;河南牧业经济学院;郑州市猪重大疫病防控重点实验室;
【基金】：河南省产学研合作资助项目(132107000002,152107000003)
【分类号】：S852.651

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