我国首株猪盖他病毒的分离与鉴定  

Isolation and Identification of the First Getah Virus (GETV) Strain HNJZ-S1 from Clinically Suspected PRRS Case of Pig Herd in Henan Province,China

作　　者：周峰[1] 崔丹丹[1] 王傲杰 王新港[1] 常洪涛[1] 陈陆[1] 李永涛[1] 王川庆[1] 

机构地区：[1]河南农业大学牧医工程学院

出　　处：《病毒学报》2018年第1期59-66,共8页Chinese Journal of Virology

基　　金：河南省高校科技创新团队支持计划(项目号:14IRTSTHN015)~~

摘　　要：盖他病毒（Getah virus,GETV）是一种广泛分布于欧亚大陆及澳大利亚北部太平洋沿岸的人兽共患虫媒病毒,我国目前各地仅从蚊子中分离到该病毒。本研究在对一猪场疑似猪繁殖与呼吸综合征（PRRS）流产胎儿进行多重RT-PCR检测时,由检测猪瘟病毒（Classical swine fever virus,CSFV）的一对引物扩增出一非特异性条带,测序及在NCBI数据库中BLAST结果显示,该序列与GETV序列高度相似。经病毒分离、传代、蚀斑纯化,获得一株能够在Marc-145细胞上稳定生长的分离物。电镜观察显示,该病毒有囊膜及纤突,直径约70nm,接近于球形,具有披膜病毒科甲病毒属成员的典型形态特征,将该分离物命名为HNJZ-S1株盖塔病毒（Getah virus）。全基因序列分析结果显示,分离物HNJZ-S1基因组全长11 689bp,与GenBank中现有14株GETV序列相似性为97.4%~99.3%,其与韩国猪源毒株AY702913（登录号AY702913）序列相似性最高（99.3%）。遗传进化分析显示,HNJZS1与日本蚊源毒株12IH26、马源毒株14-I-605-C1、14-I-605-C2及中国蚊源毒株HB0234亲缘关系最近,处于同一进化分支。其E2基因推导的氨基酸序列分析结果显示,HNJZ-S1与韩国猪源毒株AY702913和QIAG9302以及日本马源毒株14-I-605-C1、14-I-605-C2和中国蚊源毒株HB0234等处于同一进化分支。本研究首次从我国发病猪群中检测并分离到GETV,也是迄今为止中国唯一一株来自脊椎动物的盖他病毒,对猪源GETV的感染谱和致病性、感染和传播机制、环境微生物学及公共卫生等相关研究具有重要意义,并为之提供了宝贵的研究素材。Getah virus（GETV）is a zoonotic Arbovirus which is widely distributed in the Eurasian continent and northern Pacific coast of Australia.In this study,a non-specific segment was amplified by apair of primers that detected Classical swine fever virus（CSFV）when the multiple RT-PCR detection were performed on abortive fetus from a clinically suspected PRRS swine case.The result of DNA sequencing and BLAST showed that the sequence of the non-specific segment exhibited high homology with that of GETV.An acquired isolate which can stably growth on Marc-145 cells was isolated by virus passage and plaque purification.Electron microscopic observation showed that the virus has capsular and fibrous and is about 70 nm in diameter,and named strain HNJZ-S1.The results of the whole genome analysis showed that the genomic sequence of the isolate HNJZ-S1 is 11,689 nucleotides in length and shares 97.4% ~99.3% of genomic homology with full length sequences of 14 reference GETV strains previously submitted in GenBank and a higher degree of genetic homology（99.3% ）with Korean pig source strain AY702913（Accession Number.AY702913）as well.Genetic evolution analysis showed that strain HNJZ-S1 was closely related to the Japanese strain 12 IH26 originated from mosquito,horse-derived strains 14-I-605-C1 and 14-I-605-C2 and the Chinese mosquito source strain HB0234.Phylogenetic analysis based on deduced amino acids sequences of the E2 gene revealed that the strain HNJZ-S1 was closely related to Korean pig source strains AY702913 and QIAG9302 and Chinese mosquito source strains YN0540 and SH05-15.In this study,GETV was detected and isolated from diseased swine for the first time in China,and it is the only strain of GETV isolated from the vertebrates to date.The result of this study is obviously very important for researchers in the studies on all aspects of the GETV infection both in humans and animals such as the pathogenicity,infection mechanism,chain of virus transmission,environmental microbiology,public health and so on.

关 键 词：猪 流产胎儿 病毒分离 盖他病毒(GETV) 全基因序列 

分 类 号：S852.65[农业科学—基础兽医学]