猪流行性腹泻病毒PCR检测及其M基因的遗传变异分析  

PCR Detection of Porcine Epidemic Diarrhea Virus and Genetic Analysis of M Gene

作　　者：张志[1] 李岚[2] 董雅琴[1] 王赛赛[3] 刘爽[1] 吴发兴[1] 李晓成[1] 

机构地区：[1]中国动物卫生与流行病学中心,山东青岛266032 [2]云南农业大学,云南昆明650201 [3]福建农林大学,福建福州350002

出　　处：《动物医学进展》2013年第8期13-18,共6页Progress In Veterinary Medicine

基　　金：国家支撑计划项目(2007BAD86B05);广西科技攻关课题(桂科攻0815009-3-9)

摘　　要：为了解我国猪流行性腹泻病毒(PEDV)的流行情况,利用套式反转录-聚合酶链反应(nRT-PCR)技术,从表现腹泻症状的仔猪粪便中扩增出6份阳性样品,测序表明,一扩PCR产物的大小为854bp,涵盖PEDV的M基因。选择34个参考毒株基于M基因进行进化树分析,结果显示PEDV可以分为G1和G2两个基因型7个基因亚型。G1型以2007年前的经典毒株为主,包括CV777、Br1/87、KPEDV-9、JMe2和CH/S株等;G2型则以近年分离的毒株为主,包括GDXS5、JS-2004-2、PFF188、KU06RB08等。其中G2.3亚型已经成为目前世界上主要的PEDV流行毒株,包括本次检测的6株PEDV、泰国的KU06RB08及GenBank中我国和韩国2011年的流行毒株。通过同源性比较发现,这6株PEDV之间的同源性为98.2%～100%,与经典毒株CV777的同源性为97.6%～98.2%。结果表明,尽管PEDV之间的同源性仍较高,但PEDV流行毒株已经逐渐形成独特的流行进化分支。To investigate the occurrence and variation of porcine epidemic diarrhea virus（PEDV）, the nested reverse PCR was used in the detection of fecal samples from diarrhea piglets. The positive samples of first amplification were sequenced and showed 854 bp including the entire M gene. Comparing to other 34 refer- ence strains in GenBank,all PEDVs can be divided into two groups of G1 and G2 or seven subgroups of G1.1 to G2.3. G1 is consisted of classical PEDVs,such as CV777, Brl/87,KPEDV-9,JMe2 and CH/S, while G2 is consisted of epidemic PEDVs in recent years, such as GDXSS, JS-2004-2, PFF188 and KU06RB08. Phylogenetic analysis indicated that subgroup G2.3 has become the major epidemic branch of PEDVs in the world,which is composed of 6 PEDVs in this experiment, and other reference strains isola- ted from China in 2011, Korea in 2011 and Thailand in 2008. Among 6 detectable strains,they share a close relationship of 98.2%-100% ,while have an identity of 97.6%-98.2% with CV777. This results indicated that epidemic strains of PEDVs in recent years have formed a special genetic branch of G2.3 though these strains and share a highly homology.

关 键 词：猪流行性腹泻病毒 M基因 进化分析 

分 类 号：S852.65[农业科学—基础兽医学]