犬细小病毒JL-(18)1/Beagle株的分离鉴定及遗传进化分析  

Isolation and identification of canine parvovirus JL-(18) 1/Beagle strain and it’s analysis of genetic evolution

作　　者：李雪[1,4] 陈杰 胡博 陶荣珊[1,2] 刘梦佳[3] 潘虹军 龚成燕 王全凯 赵建军 LI Xue;CHEN Jie;HU Bo;TAO Rong-shan;LIU Meng-jia;PAN Hong-jun;GONG Cheng-yan;WANG Quan-kai;ZHAO Jian-jun(College of Chinese Medicine Materials,Jilin Agricultural University,Changchun 130118,China;Institute of Special Animal and Plant Science,Chinese Academy of Agricultural Sciences,Changchun 130112,China;Jinan Customs District,Jinan 250200,China;College of Animal Science and Veterinary,Heilongjiang Bayi Agricultural University,Daqing 163319,China)

机构地区：[1]吉林农业大学中药材学院,吉林长春130118 [2]中国农业科学院,特产研究所,吉林长春130112 [3]济南海关,山东济南250200 [4]黑龙江八一农垦大学动物科技学院,黑龙江大庆163319

出　　处：《中国兽医科学》2021年第1期81-89,共9页Chinese Veterinary Science

基　　金：国家重点研发计划项目(2017YFD0501600);黑龙江八一农垦大学引进人才科研启动计划项目(XYB201912);家畜疫病病原生物学国家重点实验室开放课题项目(SKLVEB2018KFKT014);国家自然科学基金项目(31972714)。

摘　　要：为研究犬细小病毒(CPV)流行株遗传变异情况,本研究从吉林省某比格犬养殖场采集疑似CPV感染致死犬的肠道组织,将病料接种至F81细胞进行病毒分离,并通过电镜形态学、PCR、血凝试验及动物回归试验进行鉴定。结果显示,所分离的这株病毒在F81细胞中产生了典型CPV细胞病变(CPE);在电镜下可见病毒呈无囊膜、直径为20 nm左右的病毒粒子;PCR鉴定证实CPV核酸呈阳性;血凝试验表明此病毒对猪红细胞具有凝集作用,血凝效价为1∶256;将这株病毒命名为JL-(18)1/Beagle。动物回归试验表明,JL-(18)1/Beagle使比格犬产生犬细小病毒感染的典型临床症状;以NS1基因进行遗传进化分析显示,JL-(18)1/Beagle与CPV-2c型分离株CPV-SH1516以及Canine/China/23/2017 NS1基因亲缘关系较近,而核苷酸相似率为100%;以VP2基因进行遗传进化分析显示,JL-(18)1/Beagle与New CPV-2b型CPV/CN/JL3/2013、CPV/CN/HB1/2013和CPV/CN/JL6/2013亲缘关系较近,核苷酸相似率为99.89%~99.94%;以VP2蛋白氨基酸进行序列分析表明,JL-(18)1/Beagle属于New CPV-2b型,与近期吉林CPV分离株相比,关键氨基酸位点无明显变化。因此,本试验从吉林省比格犬肠组织中分离到1株New CPV-2b型CPV毒株。In order to study the genetic variation of current canine parvovirus(CPV),the intestinal tissues were collected from a suspected CPV infected beagle dog in Jilin Province.The tissue sample was inoculated into F81 cells to isolate virus,and then was identified by electron morphology,PCR,hemagglutination assay and animal regression experiment.The results indicated that the virus produced typical CPE of CPV in F81 cells.The virus particles without capsule and a diameter of about 20 nm were observed by electron microscope.PCR results showed that the virus was CPV positive.The hemagglutination assay indicated that the virus was agglutinated porcine erythrocyte and the HA blood coagulation rate was 1∶256.The virus was identified to be the CPV and was named with JL-(18)1/Beagle.The animal regression experiment indicated that JL-(18)1/Beagle caused typical clinical symptoms of CPV infection in beagle dogs.The genetic evolution analysis indicated that,JL-(18)1/Beagle had a closer relationship with CPV-2 c subtype CPV-SH1516 and Canine/China/23/2017 based on NS1 gene,and nucleotide identities were 100%,JL-(18)1/Beagle had a closer relationship with CPV isolated of New CPV-2 b subtype,CPV/CN/JL3/2013,CPV/CN/HB1/2013 and CPV/CN/JL6/2013 based on VP2 gene,whose nucleotide identities were from 99.89%to 99.94%.The analysis of VP2 protein amino acid sequence suggested that JL-(18)1/Beagle belonged to New CPV-2 b subtype.The major amino acid sites of JL-(18)1/Beagle were not significantly different with the recent CPV strains of Jilin Province.In conclusion,a CPV strain of New CPV-2 b subtype was isolated from the intestinal tissue of a beagle dog from Jilin Province.

关 键 词：犬细小病毒 分离鉴定 遗传进化 

分 类 号：S852.659.6[农业科学—基础兽医学]