貉狐源阿留申病病毒的感染检测与基因特征  

Detection of raccoon dog and fox amdoparvovirus infection and viral genetic characteristics

作　　者：邵西群[1] 章秀婷[1] 于淼[1] 贾博寅[1] 岳志刚[1] 杨福合[1] 

机构地区：[1]中国农业科学院特产研究所特种经济动物分子生物学国家重点实验室,吉林长春130112

出　　处：《中国兽医科学》2015年第6期589-595,共7页Chinese Veterinary Science

基　　金：公益性行业(农业)科研专项(200903014);中国农业科学院科技创新工程2014

摘　　要：为检测新发现的貉狐阿留申病病毒(RFAV)感染并分析该病毒的基因特征,应用PCR、对流免疫电泳(CIEP)和碘凝集(IAT)试验检测RFAV感染的貉、狐样品;用阿留申病病毒属保守引物对RFAV进行PCR扩增、测序并进行序列分析。结果显示,2012至2014年77只可疑RFAV感染病兽的PCR检测阳性率为81.8%,其中58份PCR阳性血清用AMDV-G抗原的CIEP检测的阳性率是100%,IAT检测的阳性率为92.0%。而健康貉、狐用PCR、CIEP和IAT检测均为阴性。RFAV的4个代表毒株蛋白NS1、VP2氨基酸序列与水貂阿留申病病毒(AMDV)的相似性分别小于76.7%、92.1%,与同属的灰狐阿留申病病毒(GFADV)的相似性更低。与AMDV和GFADV毒株相比,RFAV的NS3蛋白序列较短,为66aa;VP2蛋白的半胱天冬酶裂解位点417DTLS/A421是S420,不同于其他2种病毒的D420;NS1蛋白的半胱天冬酶裂解位点226EE/T228,与其他2种病毒不同。所有测序的RFAV高变区核酸序列进化树显示,RFAV形成阿留申病病毒属的一个新分支。3种方法检测结果分析表明,AMDV-G毒株抗原适用于狐、貉RFAV感染后抗体的CIEP检测;IAT试验可用于RFAV导致的貉高γ球蛋白血症检测。RFAV基因特征表明其不同于水貂阿留申病病毒,是自然感染貉和蓝狐的新种阿留申病病毒;这提示应采取有效措施预防貉、狐的阿留申病病毒感染。The study was to examine raccoon dog and fox amdoparvovirus(RFAV)infection and characterize RFAV genetic traits.Three methods including PCR,Aleutian mink disease virus(AMDV)-specific counter immunoelectrophoresis(CIEP)and serum iodine agglutination test(IAT)were used to examine samples of sick animals infected by RFAV.RFAV fragment was amplified by PCR using the conserved primers within the genus Amdoparvovirus.And,the purified PCR products were sequenced directly by Sanger method.Then,RFAV sequences obtained were used to perform bioinformatics analysis.During 2012 to 2014,the positive rate of RFAV accounted for 81.8% among 77 suspected sick animals.Fifty-eight animals were both RFAV PCR and AMDV-G CIEP positive in their available sera testing.In addition,there was a high IAT positive rate(92.0%)in RFAV PCR positive blood.Meanwhile,the samples of healthy raccon dogs and foxes were negative by PCR,CIEP and IAT testing.Four representative strains of RFAV share a similarity of less than 76.7% and 92.1%in NS1 and VP2amino acid sequences of AMDV,respectively.Moreover,the four RFAV strains have lower similarity in VP2 and NS1protein with gray fox amdoparvovirus(GFADV)than AMDV.The NS3 sequences of RFAV are shorter than AMDV and GFADV of two known species in genus Amdoparvovirus.The S420(417DTLS/A421)of RFAV differs from the D420 of the AMDV and GFADV at a caspase cleavage site of VP2 protein.Additionally,the 226 E E/T228 of RFAV is distinct from those of AMDV and GFADV at a caspase cleavage site of NS1 protein.A neighbor-joining tree of the VP2 hypervariable region indicated that all RFAV isolates cluster into a unique clade between AMDV and GFADV species.The testing results indicate CIEP test can detect RFAV infection with AMDV-G antigen,and IAT can be used to examine the highγ-globulin content caused by RFAV infection in raccoon dog or fox serum.The RFAV molecular features reveal that RFAV is a new amdoparvovirus species genetically distinct from AMDV,and naturally infects farmed raccoon dogs and arctic foxes,sugges

关 键 词：貉狐阿留申病病毒 感染 检测 基因特征 

分 类 号：S858.92[农业科学—临床兽医学]