猪流行性腹泻病毒德州株的分离鉴定及ORF3基因遗传进化分析

Isolation and Identification of PEDV Dezhou Strain and Genetic Evolution Analysis of ORF3 Gene

为了分析猪流行性腹泻病毒(PEDV)的ORF 3基因遗传情况,本试验采集患有腹泻仔猪的小肠组织内容物样品12份,通过病毒分离、形态学观察、反转录PCR(RT-PCR)方法和间接免疫荧光(IFA)试验进行PEDV鉴定,并分析分离株的ORF 3基因核苷酸序列。结果显示,分离得到1株PEDV,命名为德州株(JIAOZUO101),该毒株的ORF 3基因与GenBank中登录的20株PEDV参照株ORF3基因核苷酸序列的同源性介于97.6%~99.9%,与Jiangxi 2017株、Henan 2015株位于同一支且遗传距离较近,其同源性介于99.3%~99.9%,与其他参考株的遗传距离较远;德州株(JIAOZUO101)的ORF 3基因序列中只是个别碱基发生变化,未发生明显变异。结果表明,分离获得PEDV德州株(JIAOZUO101)的ORF 3基因序列与Jiangxi 2017株、Henan 2015株同源性较近,且未发生变异情况。本试验为进一步研究PEDV的流行病学提供分子生物学依据。

In order to analyze the ORF3 gene inheritance of porcine epidemic diarrhea virus(PEDV),this study collected 12 intestinal tissue contents from piglets with diarrhea and tested for PEDV by Vero cell virus isolation,morphological observation,reverse transcription PCR(RT-PCR)and indirect immunofluorescence assay(IFA),and ORF3 gene sequencing.The results showed that one PEDV strain was isolated and named as Dezhou strain(JIAOZUO101);the ORF 3 gene of this strain had 97.6%-99.9%homology with that of 20 PEDV reference strains registered in GenBank,and 99.3%-99.9%homology with that of Jiangxi 2017 strain and Henan 2015 strain in the same branch,indicating certain evolutionary distance from other reference strains;and the ORF 3 gene sequence of Dezhou strain(JIAOZUO101)contained only individual base changes without other obvious variation.Thus,the ORF 3 gene sequence of the isolated PEDV Dezhou strain(JIAOZUO101)shares close homology with Jiangxi 2017 strain and Henan 2015 strain,and demonstrates no obvious variation.This data provides molecular evidence on the epidemiology of PEDV.