摘要
目的对2021年江西省高安市采集的鼠肺样本进行肾综合征出血热汉坦病毒分离鉴定及全基因组序列进行分析。方法采用Vero-E6细胞分离肾综合征出血热汉坦病毒,采用实时荧光RT-PCR法和直接免疫荧光法对分离株进行鉴定,采用高通量测序技术对分离株进行全基因组测序,采用CLC、MEGA7.0等软件对基因组序列进行分析。结果从褐家鼠和罗赛鼠各分离到1株汉城病毒(Seoul orthohantavirus,SEOV),分别命名为GAW30/2021、GAW50/2021。2株分离株的3个基因片段大小相同,每个基因编码区长度一致。分离株S、M、L基因分别含1773、3651、6530个核苷酸,分别编码429、1133、2151个氨基酸。2株分离株3个基因核苷酸序列同源性分别为99.7%、99.0%、99.4%、氨基酸序列同源性分别为99.8%、99.7%、99.9%。2株分离株与国内外SEOV参考株全基因组核苷酸同源性为84.3%~94.9%,氨基酸同源性为96.1%~99.8%,在系统进化树上独立分枝。2株分离株与SEOV疫苗株基因组氨基酸同源性为97.6%~99.1%;核蛋白和糖蛋白免疫表位氨基酸序列与疫苗株一致。结论首次在江西省高安市鼠肺样本中分离到2株SEOV变异株,目前疫苗对变异株仍具有保护力。
This study aimed to analyze whole genome sequences of Seoul orthohantavirus from rat lung samples collected in Gaoan City,Jiangxi Province in 2021.Vero-E6 cells were used to isolate orthohantavirus;real-time fluorescent RT-PCR and direct immunofluorescence were used to identify the isolates,high-throughput sequencing technology was used to sequence the whole genomes of the isolates,and CLC and MEGA7.0 software were used to analyze the genome sequence.Two strains of SEOV,named GAW30/2021 and GAW50/2021,were isolated from Rattus norvegicus and Rattus rosenbergii,respectively.The three gene fragments of the two strains were the same size,and the length of the coding region of each gene was the same.The S,M and L genes of the isolates contained 1773,3651 and 6530 nucleotides,encoding 429,1133 and 2151 amino acids,respectively.The S,M and L nucleotide sequence identity of the two isolates was 99.7%,99.0%and 99.4%,and the amino acid sequence identity was 99.8%,99.7%and 99.9%,respectively.The two strains shared 84.3%-94.9%nucleotide sequence identity and 96.1%-99.8%amino acid sequence identity with SEOV reference strains.The amino acid sequence identity between the two strains and the SEOV vaccine strain was 97.6%-99.1%.The amino acid sequence of nucleoprotein and glycoprotein epitopes of the two strains was consistent with that of the vaccine strain.This study reports the first isolation of SEOV strains from rat lung samples in Gaoan City,Jiangxi Province,and indicates that the current vaccine still has protective effects against variants.
作者
刘师文
张艳妮
肖芳
周珺
刘晓庆
熊英
吴杨博文
龚甜
LIU Shi-wen;ZHANG Yan-ni;XIAO Fang;ZHOU Jun;LIU Xiao-qing;XIONG Ying;WU Yang-bo-wen;GONG Tian(Jiangxi Center for Disease Control and Prevention,Nanchang 330029,China)
出处
《中国人兽共患病学报》
CAS
CSCD
北大核心
2022年第2期102-107,共6页
Chinese Journal of Zoonoses
基金
江西省科技计划项目重点研发计划(No.20202BBGL73052、No.20203BBG73055)。
关键词
汉城病毒
病毒分离与鉴定
全基因组序列
疫苗
Seoul orthohantavirus
virus isolation and identification
complete genome sequence
vaccine
