猪流行性腹泻病毒山东SD20BZ01株的分离鉴定及遗传进化分析
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Isolation,Identification and Genetic Evolution Analysis of Porcine Epidemic Diarrhea Virus SD20BZ01 Strain in Shandong Province
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    摘要:

    为了解山东省猪流行性腹泻病毒(porcine epidemic diarrhea virus,PEDV)基因组遗传演化特征,将山东省滨州市某猪场的PEDV阳性样品接种Vero E6细胞,结果成功分离到1株PEDV,将其命名为SD20BZ01株。通过RT-PCR分段扩增病毒基因组,经测序、拼接获得SD20BZ01株全基因组序列,并将其与GenBank收录的参考毒株序列进行同源性比对和遗传进化分析。全基因组序列分析结果显示,SD20BZ01株与我国2010年以来流行的GII-b亚群变异株序列同源性为97.8%~98.7%,明显高于CV777、DR13、SM98、SD-M等早期经典毒株,因此SD20BZ01属于GII-b亚群变异株。基于S基因的遗传进化关系与基于全基因组序列的遗传进化关系基本一致。与CV777株相比,SD20BZ01株S蛋白在中和抗原表位COE处有10个氨基酸突变,与近年来变异株S蛋白的氨基酸序列基本一致,说明GII-b亚群PEDV的变异规律逐渐趋于稳定。本研究为山东省PEDV流行病学研究及其流行控制策略的制定提供了理论依据。

    Abstract:

    In order to investigate the genetic evolution characteristics of porcine epidemic diarrhea virus(PEDV)in Shandong Province,PEDV-positive samples collected from a swine farm in Binzhou City of Shandong Province were inoculated with Vero E6 cells,then a strain of PEDV was successfully isolated,then named as SD20BZ01. Viral genome fragments were amplified by RT-PCR to obtain its whole genome sequence through sequencing and splicing,then the homology and genetic evolution relationship were compared with the reference strain registered in GenBank. It was found that,by the analysis of whole genome sequence,the homology of SD20BZ01 and GII-b variants prevalent in China since 2010 was from 97.8% to 98.7%,which was significantly higher than early classical strains like CV777,DR13,SM98 and SD-M. It was determined that SD20BZ01 was belonged to GII-b subtype variants. The genetic evolutionary relationship based on S gene was generally consistent with that based on the whole genome sequence. Compared with CV777,the S protein of SD20BZ01 harbored 10 amino acid mutations in the neutralizing antigen epitope,while was basically consistent with the amino acid sequence of S protein of variants prevalent in recent years,indicating that the rules of variation of GII-b subtype PEDV tended to be stable gradually. A theoretical basis was thus provided for future epidemiological research as well as development of prevention and control strategy against PEDV in Shandong Province.

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田似报,尹明荣,王文文,厉磊,王一新,李阳.猪流行性腹泻病毒山东SD20BZ01株的分离鉴定及遗传进化分析[J].《中国动物检疫》编辑部,2022,39(6):24-29.

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