四川彭州猪流行性腹泻流行株全基因序列测定及其遗传变异分析

doi:10.16431/j.cnki.1671-7236.2020.01.025

摘要/Abstract

摘要： 为了对四川省彭州某免疫过猪流行性腹泻病毒（PEDV）疫苗猪场猪感染的PEDV毒株基因组特征和遗传变异规律进行研究，本试验采用RT-PCR法筛选PEDV阳性样本，命名为SCXM株，对全基因组序列克隆测序获取全基因组序列，并对主要结构基因进行遗传变异分析和系统进化分析。结果显示，SCXM株的遗传变异主要集中在S基因，与71个毒株相比同源性为90.4%~99.2%，与猪场免疫疫苗株CV777相比同源性仅为93.8%。与国内常用疫苗毒株相比，在5个线性抗原表位（P1、S1P2、S1P3、SS5和SS6）和1个中和抗原表位（SID）均存在氨基酸位点突变。遗传进化分析表明，SCXM株属于G2b亚型PEDV，与湖北HBXY3株和越南毒株同在一个分支，表明这些地区流行毒株的演化过程存在相关性。另外对ORF3、M和N基因分析结果显示，SCXM与疫苗株相比均存在一定数量的氨基酸突变，但变异程度相对较小，且未引起抗原性预测值的变化。结果表明，PEDV SCXM株属新型变异毒株，其S基因发生较大程度的变异，S基因多个氨基酸突变和抗原性的变化可能是导致猪场疫苗免疫失败的原因之一。本研究丰富了PEDV基因学研究资料，探讨了SCXM株PEDV遗传变异和演化特征，为PEDV的分子演化研究奠定了基础。

关键词: 猪流行性腹泻病毒(PEDV); 基因组; 结构基因; 免疫失败

Abstract: In order to study the genomic characteristics and genetic variation of porcine epidemic diarrhea virus (PEDV) strains infected by a pig farm immunized with PEDV vaccine in Pengzhou,Sichuan province,RT-PCR was used to screen the positive samples of PEDV,named SCXM strain.The whole genome sequence was cloned and sequenced to obtain the whole genome sequence,and the main structural genes were analyzed for genetic variation and phylogenetic analysis.The results showed that the genetic variation of SCXM strain was mainly concentrated in S gene,the homologies were 90.4%-99.2% compared with 71 strains and 93.8% compared with CV777.Compared with common vaccine strains in China,amino acid mutations were found in 5 linear epitopes (P1,S1P2,S1P3,SS5 and SS6) and 1 neutralizing epitope (SID).The analysis of genetic evolution showed that SCXM strain belonged to the G2b subtype of PEDV,which was the same branch with HBXY3 strain in Hubei province and Vietnam strain.In addition,the analysis of ORF3,M and N genes showed that there were a certain number of amino acid mutations in SCXM compared with the vaccine strain,but the degree of variation was relatively small,and did not cause the change of antigenicity prediction value.The results showed that PEDV SCXM strain was a new variant strain,and its S gene was mutated to a large extent.The mutation and antigenicity change of multiple amino acids in S gene might be one of the reasons for the failure of vaccine immunization in pig farms.This study enriched the genetic data of PEDV,discussed the genetic variation and evolution characteristics of PEDV of SCXM strain,and laid a foundation for the molecular evolution of PEDV.

Key words: porcine epidemic diarrhea virus (PEDV); genome; structural gene; failure immunization

中图分类号:

S852.65

向萌, 张焕容, 任玉鹏, 张可可. 四川彭州猪流行性腹泻流行株全基因序列测定及其遗传变异分析[J]. 中国畜牧兽医, 2020, 47(1): 209-220.

XIANG Meng, ZHANG Huanrong, REN Yupeng, ZHANG Keke. Sequencing and Genetic Variation Analysis of the Whole Gene of Epidemic Strain of PEDV in Pengzhou,Sichuan Province[J]. China Animal Husbandry and Veterinary Medicine, 2020, 47(1): 209-220.

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