| 1,781 | 154 | 227 |
| 下载次数 | 被引频次 | 阅读次数 |
2010年底开始,猪流行性腹泻(Porcine epidemic diarrhea,PED)在中国境内出现严重暴发。为探讨该病突然再次暴发的原因,对2010年10月至2012年6月于华中地区11个地市收集的12株猪流行性腹泻病毒(Porcineepidemic diarrhea virus,PEDV)S、M和ORF3基因进行RT-PCR扩增、克隆及测序,并对其序列变化、遗传进化关系、抗原位点、糖基化位点和多肽极性进行分析。结果显示,12株PEDV流行毒株S、M和ORF3基因部分核苷酸及氨基酸的改变使其关键序列明显不同于以往毒株,其中有11株流行毒株的S基因比现用疫苗毒株CV777多9个核苷酸,而且在其S1区N′端存在大量的氨基酸突变。遗传进化分析显示,12株PEDV华中地区毒株与2007～2009年韩国毒株、2007～2008年泰国毒株、2009～2010年越南毒株、2011年日本毒株及2010～2012年中国其它地区流行毒株亲缘关系均较密切,而与欧洲株(CV 777、Br1/87)、中国现用疫苗株(CV777)及2003～2007年中国分离株(LJB/03、JS-2004-2、DX、QH、LZC)亲缘关系均较远。与现用疫苗株(CV777)相比,流行毒株S蛋白的抗原位点、糖基化位点和跨膜螺旋均有明显改变,提示PEDV在近年呈现快速变异和进化的趋势,因而可能需要选择更有效的疫苗株来控制PEDV的暴发。
Abstract：Since late 2010,porcine epidemic diarrhea virus(PEDV) has been re-emerging in central China.To explore the possible reason of the PEDV outbreaks,twelve PEDV field strains were isolated from different swine breeding farms in central China during 2010-2012,and molecular diversity,phylogenetic relationships of these strains with other PEDV reference strains were investigated.Sequence analysis of S,M and ORE3 genes revealed that the central China PEDV isolates had several specific nucleotides and amino acids which were different from PEDV reference strains.In addition,the entire S genes of eleven central China PEDV isolates were found to be nine nucleotides longer in length than CV777 and large number of amino acid variations was accumulated in the N-terminal region of S gene.Phylogenetic analysis showed that the central China PEDV isolates had close relationship with Korea strains(2007-2009),Thailand strains(2007-2008),Vietnam strains(2009-2010),Japan strains(2010),and other prevailing strains from other parts of China(2010-2012).However,they differed genetically from European strains(CV777,Br1/87),China strains(2003-2007) and the vaccine strains(CV777) used in China.These results imply that a rapid variation and evolution of central China PEDV strains has occurred in recent years,and a more efficient vaccine strain should be selected to prevent and control outbreaks of PEDV in China.
[1]Debouck P,Pensaert M.Experimental infection of pigswith a new porcine enteric coronavirus,CV777[J].Am JVet Res,1980,41(2):219-223.
[2]Debouck P,Pensaert M,Coussement W.The pathogene-sis of an enteric infection in pigs,experimentally inducedby the coronavirus-like agent,CV777[J].Vet Microbi-ol,1981,6(2):157-165.
[3]Pensaert M B,Debouck P.A new coronavirus-like parti-cle associated with diarrhea in swine[J].Arch Virol,1978,58(3):243-247.
[4]Chasey D,Cartwright S F.Virus-like particles associat-ed with porcine epidemic diarrhea[J].Res vet Sci,1978,25(2):255-256.
[5]宣华,邢德坤,王殿瀛,等.应用猪胎肠单层细胞培养猪流行性腹泻病毒的研究[J].兽医大学学报,1984,4(3):202-208.
[6]Chen J F,Sun D B,Wang C B,et al.Molecular charac-terization and phylogenetic analysis of membrane proteingenes of porcine epidemic diarrhea virus isolates in China[J].Virus Genes,2008,36(2):355-364.
[7]Puranaveja S,Poolperm P,Lertwatcharasarakul P,et al.Chinese-like strain of porcine epidemic diarrhea virus,Thailand[J].Emerg Infect Dis,2009,15(7):1112-1115.
[8]Park S J,Kim H K,Song D S,et al.Molecular charac-terization and phylogenetic analysis of porcine epidemicdiarrhea virus(PEDV)field isolates in Korea[J].ArchVirol,2011,156(4):577-585.
[9]Duy D T,Toan N T,Puranaveja S,et al.Genetic charac-terization of porcine epidemic diarrhea virus(PEDV)isolates from Southern Vietnam during 2009-2010outbreaks[J].Thai J Vet Med,2011,41(1):55-64.
[10]Kocherhans R,Bridgen A,Ackermann M,et al.Completion of the porcine epidemic diarrhea corona-virus(PEDV)genome sequence[J].Virus Genes,2001,23(2):137-144.
[11]Jackwood M W,Hilt D A,Callison S A,et al.Spikeglycoprotein cleavage recognition site analysis ofinfectious bronchitis virus[J].Avian Dis,2001,45(2):366-372.
[12]Sturman L S,Holmes K V.Proteolytic cleavage ofpeplomeric glycoprotein E2of MHV yields two 90Ksubunits and activates cell fusion[J].Adv Exp MedBiol,1984,173:25-35.
[13]Duarte M,Laude H.Sequence of the spike protein ofthe porcine epidemic diarrhoea virus[J].J Gen Virol,1994,75:1195-1200.
[14]Cruz D J M,Kim C J,Shin H J.The GPRLQPY motiflocated at the carboxy-terminal of the spike proteininduces antibodies that neutralize Porcine epidemicdiarrhea virus[J].Virus Res,2008,132(1-2):192-196.
[15]Sun D B,Feng L,Shi H Y,et al.Identification of twonovel B cell epitopes on porcine epidemic diarrheavirus spike protein[J].Vet Microbiol,2008,131(1-2):73-81.
[16]Ballesteros M L,Sanchez C M,Enjuanes L.Two aminoacid changes at the N-terminus of transmissible gas-troenteritis coronavirus spike protein result in the lossof enteric tropism[J].Virology,1997,227:378-388.
[17]Kwon H M,Saif L J,Jackwood D J.Field isolates oftransmissible gastroenteritis virus differ at the molec-ular level from the Miller and Purdue virulent and at-tenuated strains and from porcine respiratory corona-virus[J].J vet med sci,1998,60(5):589-597.
[18]Woods R D.Efficacy of a transmissible gastroenteritiscoronavirus with an altered ORF3gene[J].Can JVet Res,2001,65(1):28-32.
[19]Song D S,Yang J S,Oh J S,et al.Differentiation of avero cell adapted porcine epidemic diarrhea virus fromKorean field strains by restriction fragment lengthpolymorphism analysis of ORF3[J].Vaccine,2003,21(17-18):1833-1842.
[20]Chen J F,Wang C B,Shi H Y,et al.Molecular epide-miology of porcine epidemic diarrhea virus in China[J].Arch Virol,2010,155(9):1471-1476.
[21]Sun R Q,Cai R J,Chen Y Q,et al.Outbreak of porcineepidemic diarrhea in suckling piglets,China[J].Emerg Infect Dis,2012,18(1):161-163.
基本信息:
DOI：10.13242/j.cnki.bingduxuebao.002381
中图分类号:S852.65
引用信息:
[1]郑逢梅,霍金耀,赵军,等.2010～2012年华中地区猪流行性腹泻病毒分子特征和遗传进化分析[J].病毒学报,2013,29(02):197-205.DOI:10.13242/j.cnki.bingduxuebao.002381.
2013-03-29
2013-03-29
2013-03-29