【摘要】 目的对2007年从浙江省丽水地区肾综合征出血热(hemorrhagic fever with renal syndrome,HFRS)疫区鼠肺分离的汉坦病毒进行型别鉴定和分子生物学特性研究。方法将汉坦病毒抗原阳性的鼠肺标本接种Vero-E6细胞,分离汉坦病毒。提取病毒总RNA,应用逆转录聚合酶链反应(RT-PCR)扩增病毒S基因全片段,克隆入质粒载体,进行核苷酸序列测定及分析,应用DNA STAR软件分析比较,确定病毒型别。结果从疫区7份阳性鼠肺中分离到2株汉坦病毒,经汉坦病毒单克隆直接荧光血清检查和基因序列测定,结果为汉滩型(HTN)病毒,2株病毒与国际标准株76-118(HTN型)和80-39(SEO型)S片段核苷酸的同源性分别为84.7%、84.6%和64.2%、64.0%。结论浙江省丽水地区可能存在以HTN型病毒为主的肾综合征出血热自然疫源地。更多还原

【Abstract】 The Hantaviruses were isolated from mouse lung specimens by inoculating the positive mouse lung samples into Vero-E6 in the epidemic area of Lishui district of Zhejiang province where the infection of hemorrhagic fever with renal syndrome(HFRS)prevailed.Then,the serotype of viruses isolated was identified and their molecular biological characteristics were investigated.The total RNA of viruses isolated was extracted from the Hantavirus-infected Vero-E6 cells and the S-segment cDNA of Hantavirus was obtained by RT-PCR.This gene fragment was then cloned into plasmid vector pUCm-T and sequenced afterwards by using the DNA STAR software for comparison.It was found that two strains of Hantaviruses were isolated from 7 samples of positive mouse lungs and identified by direct immunofluorescent assay with monoclonal antibodies against Hantavirus and sequence analysis.In comparison with the international standard strains of Hantavirus 76-118(HTN)and 80-39(SEO),the homologies among them were 87.4%,84.6%,64.2% and 64.0% respectively.It is evident that nature prevalence area of HFRS could exist in Lishui district of Zhejiang province,in which the HTN strain of Hantavirus might be the predominant serotype of viruses harbored.更多还原