摘要
脑心肌炎病毒（Encephalomyocarditis virus,EMCV）是一种自然疫源性人兽共患病病原,但有关地方土猪EMCV的分离、鉴定及全基因组研究等尚未见报道。本研究应用＂细胞接种与RT-PCR方法相结合＂技术,成功分离到国内首株淮南猪源EMCV,命名为HNXX13株,并对其进行了系统鉴定和全基因组序列测定分析。结果显示,该病毒粒子大小约为24~30nm,呈圆形,不耐酸和热,对胰蛋白酶敏感,对氯仿不敏感,二价阳离子没有保护作用,所感染BHK-21细胞的细胞浆内可见到特异的绿色荧光;基因组全长为7 725bp（GenBank登录号：KF771002）,与不同动物源参考毒株的核苷酸同源性为81.0%~99.9%,与国内猪源参考毒株同源性为99.5%以上;基于全基因组序列和ORF的系统发育进化树显示,EMCV可分为G1、G2和G3 3个群,HNXX13株与国内其他参考毒株同属于G1群。研究结果表明,地方土猪可感染EMCV并引起发病,丰富了我国EMCV分子流行病学资料,并提醒在进行地方品种养殖中要充分考虑人兽共患疫病传播的生态学;EMCV存在较大的地域差异,其传播具有一定的区域限制性;EMCV在地方土猪和鼠之间可能存在着交叉感染与传播;EMCV在感染地方土猪时可能会发生个别氨基酸的突变,以适应不同的猪种。
Encephalomyocarditis virus（EMCV）is a natural epidemic zoonotic pathogen.However,no reports have been published regarding the isolation,identification and full-length genome of EMCV from a local aardvark population.In present study,an EMCV isolate HNXX13 was isolated from aardvarks named Huainan-pig in Henan Province.The systematic identification,full-length genome sequencing and molecular characteristic analysis of the isolate HNXX13were conducted.The result showed that the isolate was spherical with a diameter of 24-30nm,neither heat-nor acid-resistant,sensitive to trypsin,insensitive to chloroform,not protected by bivalent cationic,and the specific fluorescence was observed in the cytoplasm of BHK-21cells infected with the isolate by using indirect fluorescence assay.The full-length genome of EMCV HNXX13generated a 7 725bp sequence（GenBank：F771002）,with 81.0%-99.9%nucleotide identity to reference strains from different animals,and 99.5% with a Chinese reference strain isolated earlier from a commercial pig herd.The phylogenetic tree based on the full-length genome and ORF sequences identified that all EMCV strains were divided into three groups G1,G2and G3,and strain HNXX13belonging to the G1group with other Chinese reference strains.The result also identified that this EMCV infection could cause severe clinical signs in a local aardvark population,and enriches the molecular epidemiological data of EMCV in China.Regional differences exist in EMCV genome and transmission is limited within a certain area.However,the cross-infection and transmission of EMCV between aardvark and mice appears most likely.Mutations have occurred in some amino acids of EMCV strain HNXX13during the transmission in local aardvark herd and these mutations might make the virus easier to infect the aardvark
出处
《病毒学报》
CAS
CSCD
北大核心
2014年第4期375-381,共7页
Chinese Journal of Virology
关键词
脑心肌炎病毒
地方土猪
RT-PCR
全基因组
分子特征分析
Encephalomyocarditis virus
Local aardvark population
RT-PCR
Full-length genome
Mo-lecular analysis
