江西省汉坦病毒分离及其S和M基因特征分析

Hantavirus isolation and the complete S,M segments genetic analysis of the isolates in Jiangxi Province,China

目的对2011年江西省采集的鼠肺标本进行汉坦病毒的分离鉴定,了解分离病毒株的基因特征。方法采用Vero-E6细胞对阳性鼠肺标本进行病毒分离,采用直接免疫荧光法和RT-PCR方法对毒株进行鉴定。扩增分离株的S、M片段基因进行克隆测序。运用DNAstar软件包和MEGA3.1软件对序列进行分析。结果从15份标本分离到2株来自褐家鼠的汉城型汉坦病毒。对其中1株病毒的S、M片段进行了克隆和序列测定,其中S片段含1 772个核苷酸,编码429个氨基酸;M片段含3 651个核苷酸,编码1 133个氨基酸。经核苷酸和氨基酸同源性分析,新分离株与国内外汉城型病毒核苷酸同源性94.8%~98.0%,氨基酸同源性98.2%~99.6%。与汉城型标准株80-39株相比,S片段仅1个氨基酸位点发生变异;M片段有9个氨基酸位点发生变异,糖基化位点的数目和位置没有发生变化。结论从江西省褐家鼠分离到两株汉城型病毒,病毒基因变异较小,型别相对稳定。

We isolated Hantavirus from the rodents＇ lung samples in order to know the genetic characteristic of new isolates in Jiangxi Province.Samples were collected in 2011,and Hantavirus was isolated through Vero-E6 cells culture,the strains were identified by direct immunofluorescence assay and RT-PCR.The complete S,M segments of one strain were amplified and sequenced,the gene sequences were analyzed with DNAStar and MEGA3.1software.Two Seoul viruses（SEOV）were isolated and identified from 15 samples.The complete S segment contained 1 772 nucleotides with a single open reading frame coding429 amino acids;M segment contained 3 651 nucleotides in length with a single open reading frame coding 1 133 amino acids.The similarities to other SEOVs based on S/M nucleotides and amino acid sequences were 94.8%-98.0% and 98.2%-99.6% respectively.Compared to 80-39 strain,there was only one amino acid site changed in the S amino acid sequence and 9amino acid sites changed in the M amino acid sequence.Results showed that comparing to other SEOVs the genes of new isolates are highly conserved.