陕西省9株乙脑病毒的分离及E基因序列分析  

Isolation and E-Gene Sequencing of Nine Strains of the Japanese Encephalitis Virus from Shaanxi Province,China

作　　者：刘晓宇[1] 王艺楠 胡伟军[1] 殷启凯 张少白[1] 王环宇[2] LIU Xiaoyu;WANG Yinan;HU Weijun;YIN Qikai;ZHANG Shaobai;WANG Huanyu(Shaanxi Center for Disease Control and Prevention,Xi'an 710054,China;National Institute for Viral Disease Control and Prevention,Chinese Center for Disease Control and Prevention,Beijing 102206,China)

机构地区：[1]陕西省疾病预防控制中心,西安710054 [2]中国疾病预防控制中心病毒病预防控制所,北京102206

出　　处：《病毒学报》2022年第3期644-651,共8页Chinese Journal of Virology

基　　金：陕西省科技计划项目(项目号:2013K12-15-01),题目:陕西省虫媒病毒谱研究。

摘　　要：分析陕西省分离的9株乙脑病毒基因组序列特征。使用乙脑病毒全基因组序列测定引物进行RT-PCR扩增,扩增产物进行测序,拼接后获得基因组序列。利用MEGA 4.1、MegAlin、MEGA7.0等软件进行毒株的系统进化分析,并与P3株、减毒活疫苗SA14-14-2株及覆盖5个基因型别的其他乙脑病毒进行E基因序列比对。9株分离株3株分离自猪舍、6株分离自羊舍,其中4株获得全基因组序列,5株测得E基因序列。基于E基因序列进行毒株核苷酸、氨基酸同源性比较,结果显示分离株均与基因I型GI-b亚型毒株核苷酸和氨基酸同源性最高,核苷酸同源性范围为96.5%~99.7%、氨基酸同源性范围99.2%~100.0%;与SA14-14-2株核苷酸同源性范围为87.5%~88.9%、氨基酸同源性范围96.3%~97.2%;与P3株核苷酸同源性范围为87.6%~88.1%、氨基酸同源性范围96.7%~97.6%。分析09年(陕南地区)分离株与18年(关中地区)分离株的E基因核苷酸差异率为1.8%~2.9%、氨基酸差异率为0%~0.8%。陕西省自然界中循环的乙脑病毒以基因Ⅰ型为主,与P3株在抗原毒力关键位点无差异,病毒相对保守。病毒分子生物学特征及分离生境提示我省需注意多种类乙脑传播媒介和宿主的防控。To analyze the genome sequence of nine Japanese encephalitis virus(JEV) strains isolated from Shaanxi Province(China). Primers for JE virus whole genome sequencing were used to perform RT-PCR amplification. After sequencing and splicing of amplified products,genome sequences were obtained. MEGA4.1,MegAlin,and MEGA 7.0 were employed for phylogenetic analysis of the virus strains. The E-gene sequences of isolated strains were compared with those of the P3 strain,attenuated live vaccine SA14-14-2strain,and the other JEV strains covering five genotypes. Of the nine strains,three were isolated from a pig farm and six were isolated from a sheep barn. The whole genome sequence of four strains and E-gene sequence of five stains were obtained. The homology of nucleotides and amino acids was compared based on the E-gene sequence. All nine strains showed the highest homology for nucleotides and amino acids with genotype GI-b subtype strains. The nucleotide homology was 96.5% – 99.7%. The amino-acid homology was 99.2% –100.0%. When compared with the SA14-14-2 strain,the nucleotide homology was 87.5% – 88.9%,and the amino-acid homology was 96.3% – 97.2%. When compared with the P3 strain,the nucleotide homology was87.6%–88.1%,and the amino-acid homology was 96.7%–97.6%. Between isolates from 2009 in Guanzhong of Shaanxi Province and isolates from 2018 in southern Shaanxi,the difference in nucleotides in the E gene was1.8% – 2.9%,and that for amino acids was 0% – 0.8%.Gene type-I was the primary type of JEV in Shaanxi Province. There was no difference in the key sites of antigen virulence with the P3 strain,which suggested that the isolated viruses were relatively conserved. The molecular-biological structure and isolation habitats of viruses suggest that we should pay attention to the prevention and control of multiple vectors and hosts of the JEV.

关 键 词：乙型脑炎病毒 全基因组 系统进化 

分 类 号：R373.31[医药卫生—病原生物学]