猪流行性腹泻病毒变异株(CH/YNKM8/2013)的分离鉴定和全基因组序列分析
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国家现代产业(生猪)技术体系项目(CARS36);国家自然科学基金项目(31272572)


Isolation,identification and full length genome sequence analysis of a variant porcine epidemic diarrhea virusCH/YNKM8/2013
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    摘要:

    为研究猪流行性腹泻病毒(PEDV)变异株特性,通过细胞培养、细胞病变(CPE)观察、RTPCR、间接免疫荧光实验和透射电镜观察分离鉴定1株PEDV变异株(CH/YNKM8/2013株)并分析了全基因组序列。运用基因组分段扩增方法,测得该毒株的全基因组序列(GenBank登录号为KF761675.1)。该毒株S基因N端比CV777毒株S基因的N端长9 bp,包括15 bp的插入和6 bp的缺失,表明CH/YNKM8/2013为变异毒株。同源性分析显示,该毒株的基因组序列与我国广东2011年分离毒株LC的同源关系最近,相似性为99.6%,与韩国毒株KNU1305和美国毒株USA/Minnesota84/2013的序列相似性均为99.0%,而与CV777疫苗毒株的相似性较低,为96.7%。序列进化树分析表明该毒株和我国新近毒株、韩国及美国的变异毒株进化关系较近,而与我国之前流行的PEDV毒株以及疫苗毒株CV777处于不同分支。

    Abstract:

    To characterize the variant porcine epidemic diarrhea virus (PEDV) and lay the foundation for prevention and control of porcine epidemic diarrhea (PED),a variant PEDV strainCH/YNKM8/2013 was isolated in 2013 in China.The strain was identified by cytopathic effects (CPEs),RTPCR,IFA,and transmission electron microscope (TEM).The genome sequence of it was sequenced by segmented amplification method and was deposited at GenBank under the accession number of KF761675.1.The Nterminal of S gene of CH/YNKM8/2013 was 9 bp longer than that of CV777 strain,including 15 bp insertions and 6 bp deletions,which suggested that CH/YNKM8/2013 was a variant PEDV strain.The CH/YNKM8/2013 had high sequence similarity of 99.6% with the Chinese strain LC (isolated in 2011 in Guangdong province),of 99.0% with the Korea strain KNU1305 and the USA strain USA/Minnesota84/2013 and a lower sequence similarity of 96.7% with the vaccine strainCV777,respectively.The phylogenetic analysis showed that CH/YNKM8/2013 was clustered into the same group with Chinese,South Korea and USA variant PEDV strains,while it was clustered into different branch with former Chinese classical PEDV strains and the vaccine strainCV777.

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吴美洲,陈芳洲,李中华,万胜锋,叶十一,何启盖.猪流行性腹泻病毒变异株(CH/YNKM8/2013)的分离鉴定和全基因组序列分析[J].华中农业大学学报,2016,35(4):93-99

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  • 收稿日期:2015-11-30
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  • 在线发布日期: 2016-06-30
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