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为了分析中国部分省市犬细小病毒(Canine parvovirus,CPV)流行毒株的分子生物学特征,了解其遗传演化规律,查明中国目前CPV流行毒株现状,本项研究通过对2006~2008年武汉、南京和北京的宠物犬疑似犬细小病毒粪便样品进行检测,并对犬细小病毒PCR阳性产物进行RFLP分析,未发现宠物犬感染猫细小病毒的现象。从PCR检测犬细小病毒阳性标本15份以及广泛使用的犬细小病毒疫苗2株中克隆犬细小病毒VP2基因并进行序列分析,结果显示:检测到的15株犬细小病毒均属于CPV-2a亚型;与GenBank中的其他CPV-2a亚型相比,其中14株犬细小病毒存在独特的Ile324变异,并且国内主要使用的两个犬细小病毒疫苗株均属于CPV-2型,但是与CPV-2型原型毒株CPV-b相比,VP2蛋白氨基酸序列已经发生改变。通过构建基于犬细小病毒VP2蛋白氨基酸序列的系统发生树,发现在我国主要城市的宠物犬中流行的犬细小病毒CPV-2a亚型毒株单独构成一簇,并且与2008年的韩国分离株K001关系紧密,而与早期其他地区的犬细小病毒CPV-2a亚型流行株距离较远。这提示犬细小病毒流行株的变异具有某种时间和空间相关性。本研究对犬细小病毒流行株的主要衣壳蛋白VP2的编码蛋白进行遗传变异分析,为更好地预防和控制犬细小病毒提供基础。
Abstract：To recognize the molecular biology character,phylogenetic relationship and the state quo prevalent of Canine parvovirus(CPV),Faecal samples from pet dogs with acute enteritis in the cities of Beijing,Wuhan,and Nanjing were collected and tested for CPV by PCR and other assay between 2006 and 2008.There was no CPV to FPV(MEV) variation by PCR-RFLP analysis in all samples.The complete ORFs of VP2 genes were obtained by PCR from 15 clinical CPVs and 2 CPV vaccine strains.All amplicons were cloned and sequenced.Analysis of the VP2 sequences showed that clinical CPVs both belong to CPV-2a subtype,and could be classified into a new cluster by amino acids contrasting which contains Tyr→Ile(324) mutation.Besides the 2 CPV vaccine strains belong to CPV-2 subtype,and both of them have scattered variation in amino acids residues of VP2 protein.Construction of the phylogenetic tree based on CPV VP2 sequence showed these 15 CPV clinical strains were in close relationship with Korea strain K001 than CPV-2a isolates in other countries at early time,It is indicated that the canine parvovirus genetic variation was associated with location and time in some degree.The survey of CPV capsid protein VP2 gene provided the useful information for the identification of CPV types and understanding of their genetic relationship.
[1]Pollook R V,Coyne M J.Canine Parvovirus[J].VetClin North,1993,23(3):555-568.
[2]Appel M J G,Scott F W,Carmichael L E.Isolation andimmunization studies of a canine parvo-like virus fromdogs with haemorrhagic enteritis[J].Vet Rec,1979,105:156-159.
[3]Parrish,C R.Host range relationships and the evolutionof canine parvovirus[J].Vet Microbiol,1999,69(1-2):29-40.
[4]Parrish C R,Aquadro C F,Strassheim M L,et al.Rapid antigenic-type replacement and DNA sequence evo-lution of canine parvovirus[J].J Virol,1991,65:6544-6552.
[5]Parrish C R,Have P,Foreyt W J,et al.The globalspread and replacement of canine parvovirus strains[J].JGen Virol,1988,69(Pt.5):1111-1116.
[6]缪勤,张汇东,朱骞,等.犬细小病毒南京分离株基因型分析[J].畜牧与兽医,2005,37(6):20-21.
[7]Masami M,Motohiro H,Hisami H,et al.Isolation ofCanine Parvovirus from a Cat Manifesting Clinical Signsof Feline Panleukopenia[J].J Clin Microbiol,1996,34(9):2101-2105.
[8]梁士哲,梁川玫,魏喜仁,等.犬出血性肠炎的研究[J].上海畜牧兽医通讯,1982,2(4):172.
[9]徐汉坤,金淮,郭宝发,等.某犬群暴发犬病毒性肠炎的流行病学和临床病理学观察[J].畜牧与兽医,1983,5:8-10.
[10]Schunck B,W Kraft,U Truyen,et al.A simple tou-ch-down polymerase chain reaction for the detection ofcanine parvovirus and feline panleukpenia virus in feces[J].J Virol Methods,1995,55:427-433.
[11]易立,闫喜军,罗彬,等.水貂病毒性肠炎巢式PCR、PCR-RFLP联合诊断方法的建立[J].经济动物学报,2008,12(4):204-208.
[12]Kumar S,Tamura K,Nei M.MEGA3:Integratedsoftware for Molecular Evolutionary Genetics Analysisand sequence alignment[J].Briefings in Bioinformat-ics,2004,5:150-163.
[13]Parrish C R,O Connell P H,Evermann J F,et al.Natural variation of canine parvovirus[J].Science,1985,230:1046-1048.
[14]Nakamura M,Tohya Y,Miyazawa T,et al.A novelantigenic variant of Canine parvovirus from a Vietnam-ese dog[J].Arch Virol,2004,149:2261-2269.
[15]Ikeda Y,Mochizuki M,Naito R,et al.Predominanceof canine parvovirus(CPV)in unvaccinated cat popula-tions and emergence of new antigenic types of CPVs incats[J].Virology,2000,278:13-19.
[16]Decaro N,Martella V,Desario C,et al.First detec-tion of canine parvovirus type 2c in pups with haemor-rhagic enteritis in Spain[J].J Vet Med B Infect DisVet Public Health,2006,53:468-472.
[17]Buonavoglia C,Martella V,Pratelli A,et al.Evidencefor evolution of canine parvovirus type 2 in Italy[J].JGen Virol,2001,82:3021-3025.
[18]Campbell R G.The molecular epidemiology of canineparvovirus as an archetypal emerging infectious disease[M].Bachelor of Medical Science Honors Thesis.School of Biomedical Science.James Cook University,Townsville,2003.
[19]Agbandje M,McKenna R,Rossmann M G,et al.Structure determination of feline panleukopenia virusempty particles[J].Proteins,1993,16:155-171.
[20]Xie Q,Chapman M S.Canine parvovirus capsid struc-ture,analyzed at 2.9 resolution[J].J Mol Biol,1996,264:497-520.
[21]Seok-Young JEOUNG,So-Jeo AHN,Doo KIM.Ge-netic Analysis of VP2 Gene of Canine Parvovirus Iso-lates in Korea[J].J Vet Med Science,2008,70:719-722.
[22]Pratelli A,Cavalli A,Martella V,et al.Canine Parvo-virus(CPV)Vaccination:Comparison of NeutralizingAntibody Responses in Pups after Inoculation withCPV2 or CPV2b Modified Live Virus Vaccine[J].CVI2001,8:612-615.
基本信息:
中图分类号:S852.659.2
引用信息:
[1]易立,程世鹏,闫喜军,等.犬细小病毒VP2基因的遗传变异分析[J].病毒学报,2009,25(06):452-459.
基金信息:
吉林省科技发展计划项目(20080213)