新疆和田地区1株小鹅瘟病毒分离鉴定及VP基因的克隆分析  

Isolation and identification of a strain of goose Parvovirus and cloning analysis of VP gene in Hotan area,Xinjiang Uygur Autonomous Region

作　　者：刘建华[1] 孙文程 胡月[1] 吴桂灵 马彩红 撒瑞雪 阿力米古丽·乌米尔哈孜 LIU Jianhua;SUN Wencheng;HU Yue;WU Guiling;MA Caihong;SA Ruixue;Alimgul Wumirhaz(College of Veterinary Medicine,Xinjiang Agricultural University,Urumqi 830052,China;Xinjiang Uygur Autonomous Region Tacheng City Animal Disease Control and Diagnosis Center,Tacheng 834700,China)

机构地区：[1]新疆农业大学动物医学学院,乌鲁木齐830052 [2]新疆塔城地区动物疾病控制与诊断中心,新疆塔城834700

出　　处：《黑龙江畜牧兽医》2023年第15期84-89,共6页Heilongjiang Animal Science And veterinary Medicine

基　　金：新疆维吾尔自治区重点研发计划项目(2020B01004-2)。

摘　　要：为了找到新疆和田地区某鹅场雏鹅大量死亡的原因,试验采用PCR方法及琼脂扩散试验检测了病死鹅组织,将阳性病料经无菌处理后接种无免番鸭胚,盲传3代,进行病毒分离;设计GPV VP基因特异性引物对其尿囊液进行扩增,构建重组质粒和测序,分析VP基因核苷酸序列同源性、亲缘关系,以及VP蛋白的抗原表位。结果表明:PCR检测及琼脂扩散试验结果均为阳性,其他常见病毒PCR检测结果为阴性;接种病料组织研磨悬液后无免番鸭胚出现典型症状,最终分离到一株GPV分离株,将其命名为GPV/GOOSE/XJHT/2020。该毒株VP基因核苷酸序列与GenBank中不同国家地区毒株的相似性均在95.0%以上;与强毒株DY16、RC16核苷酸序列的相似性均为99.3%,亲缘性高且处在同一分支;与疫苗株SYG61v、弱毒株GPV-98E的相似性均为95.0%,亲缘性较低且处在不同分支。VP蛋白具多个潜在的抗原表位。说明该鹅场存在GPV感染,且与国内毒株亲缘关系较近。In order to find the cause of a large number of deaths of geese in a goose farm in Hotan area,Xinjiang Uygur Autonomous Region,the experiment used PCR detection method and agar diffusion test to detect the tissues of the diseased and dead geese in the goose farm.After aseptic treatment,the positive disease materials were inoculated into non-vaccination Muscovy duck embryos,which was blindly passed on for three generations,and the virus was isolated.Goose Parvovirus(GPV)VP gene-specific primers were designed for the amplification from its allantoic fluid,and the cloning vector was constructed and sequenced.The nucleotide sequence homology and genetic relationship of VP gene were analyzed,and the epitopes of VP protein were analyzed.The results showed that the PCR test and agar diffusion test were positive,and the PCR results of other common viruses were negative.After inoculation of diseased tissue grinding suspension,non-vaccination Muscovy duck embryos showed typical symptoms,and finally a GPV strain was isolated and named CPV/Goose/XJHT/2020.The VP gene sequence of this strain had more than 95.0%homology with different national and regional strains in GenBank.The nucleotide sequence homology between this strain and the virulent strains DY16 and RC16 were both 99.3%and the genetic relationship was high and they were in the same branch.Its homology with the vaccine strain SYC61v and attenuated strain GPV-98E was 95.0%,and the genetic relationship was low and they were in different branches.VP protein had multiple potential epitopes.The results indicated that GPV infection existed in the goose farm and was genetically related to the domestic strains.

关 键 词：小鹅瘟病毒 分离 鉴定 VP基因 进化分析 抗原表位预测 

分 类 号：S852.65.92[农业科学—基础兽医学]