中国兽医科学2024，Vol.54Issue(4)：511-518,8.DOI:10.16656/j.issn.1673-4696.2024.0064
一株羊肠道病毒的分离鉴定及其基因组特征分析
Identification and genomic characteristics of a novel caprine enterovirus stain
摘要
Abstract
In this study,we identified the mixed infection of peste des petits ruminants virus(PPRV)and CEV from the lymphoid tissue of a dead sheep by metagenome sequencing technology.To isolate CEV,the total viral RNA from the mesenteric lymph node tissue was used to transfect into BHK21 cells.After continuous passage,the virus was isolated and identified by cytopathogenic observation,sequencing,TCID50,one-step growth curve of virus,and electron microscope observation.The genetic characteristics of the isolated virus were compared with the reference strains.The results indicated that BHK21 cells transfected with total viral RNA showed obvious CPE in the second passage and sequencing showed that the culture only contained CEV,and its total genome length was 7 446 nt,which possess 5'UTR length of 821 nt,3'UTR of 109 nt and the ORF of 6 516 nt.The isolated virus was named JSNT/1/2022 strain and divided into enterovirus G(EV-G),compared with the reference strains.The virus particle was measured 2030 nm in diameter,and the TCID50 value of the 6 th generation virus was 104,2/0.1 mL at 8 h.In this study,a strain of CEV was firstly isolated by transfection of tissue material RNA into cells,indicating that the nucleic acid of CEV has infectious,which provides a more reliable method for the isolation and identification of CEV.The isolated virus will lay the etiological foundation for the basic and applied research on CEV.关键词
羊肠道病毒/分离鉴定/RNA感染性/基因组Key words
caprine enterovirus/isolation and identification/RNA infectivity/macrogenomic sequencing分类
农业科技引用本文复制引用
张雨星,白兴文,刘在新,卢曾军,曾巧英,王桂荣,孙普,马雪青,李平花,魏达礼,李冬,张婧,王卉,袁红,王萌..一株羊肠道病毒的分离鉴定及其基因组特征分析[J].中国兽医科学,2024,54(4):511-518,8.基金项目
兰州市科技计划项目(2022-2-46) （2022-2-46）
甘肃省科技重大专项(22ZD6NA001) （22ZD6NA001）
