雏鸭坦布苏病毒分离鉴定及全基因序列分析  

Isolation and Genomic Sequence Analysis of Duck Tembusu Virus

作　　者：提金凤 李志杰 李舫 

机构地区：[1]山东畜牧兽医职业学院,山东潍坊261061

出　　处：《动物医学进展》2017年第9期128-133,共6页Progress In Veterinary Medicine

基　　金：山东省高等学校科技发展计划项目(J16LF58);潍坊市科技发展计划项目(2013YD139;20121366);山东省现代农业产业技术体系家禽创新团队建设项目(SDAIT-13-011-05)

摘　　要：山东潍坊地区某鸭场养殖的21日龄樱桃谷鸭突然发病,病鸭主要表现翅腿瘫痪等神经症状,排黄绿色或黄白色稀粪,病死率5%左右。为了确定发病鸭群感染病原的种类,首先采集病死鸭肝脏,处理后接种9日龄~11日龄SPF鸡胚进行病原分离。其次,对分离的病原RT-PCR扩增和序列测定,MEGA5绘制系统发育进化树,DNA Star软件分析核苷酸同源性。结果表明,通过SPF鸡胚分离到一株坦布苏病毒(TMUV),命名为TMUV-SDWF。该病毒株基因组全长为10 990nt,编码1个含3 426个氨基酸的多聚蛋白。系统发育进化树分析显示,TMUV-SDAG毒株属于Ⅰa分支。核苷酸同源性分析表明,TMUV-SDSG与GenBank上公布的16株不同TMUV分离株核苷酸同源性高达97.0%~99.8%。成功分离鉴定一株鸭坦布苏病毒并对其进行了遗传进化分析,为鸭坦布苏病毒弱毒或灭活疫苗的筛选制备及该病毒分子流行病学的研究奠定了基础。Cherry valley ducks at 21 days old got sick suddenly in Shandong Weifang and the ill ducks ex-pressed neurological symptoms of paralysis, yellow-green or yellowish-white loose stool, and mortality a-bout 5%. In order to determine the pathogen of sick ducks,firstly dead duck livers were collected and used to isolate and identify the pathogen through 9-11 day-old SPF chicken embryoes. Secondly the whole ge-nome of virus was amplified by RT-PCR and sequenced. The phylogenetic tree was drawn by MEGA5 soft-ware and the nucleotide homology analysis was carried out by DNA Star software. The result showed that one Tembusu virus （TMUV） strain was isolated by SPF chicken embryo and was named TMUV-SDSG. The whole genome is 10 990 nt and encodes a polyprotein containing 3 426 amino acids. The analysis of phylogenetic tree showed that the TMUV-SDAG strain belongs to I a branch. The nucleotide homology a-nalysis results showed that the nucleotide homology is as high as 97. 0%-99. 8% among the TMUV-SDSG strain and 16 TMUV isolates in GenBank. A TMUV strain was isolated successfully and analyzed which laid the foundation for screening and preparation of attenuated and inactivated vaccines and molecular epi-demiological study.

关 键 词：鸭坦布苏病毒 核苷酸同源性分析 多聚蛋白 

分 类 号：S852.659.6[农业科学—基础兽医学] S858.32[农业科学—兽医学]