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目的 分离辽宁地区汉坦病毒并对分离株 (SY 13)进行基因分型 ,以查清辽宁地区汉坦病毒流行株的基因型和基因亚型。方法 采用组织细胞培养法分离汉坦病毒 ;用RT PCR法分别扩增SY 13的M片段G1、G2区和S片段 ;采用邻位相连法 ,通过DNAStar、Clustalx、Phylip等序列分析软件分别对 3个基因片段的序列与从GenBank下载的序列进行同源性分析 ,并构建种系发生树。结果 通过对 3个基因片段进行比较分析 ,SY 13与HTN型同源性较高 ,为 79.3%～ 97.0 % ;通过M片段G2区的比较分析 ,SY 13与BAO 10、BAO 14、JIANG 13、BAO 9、Q 33处于同一分支 ,同源性为 95.0 %～97.0 %。结论 辽宁地区汉坦病毒SY 13株与黑龙江地区分离株属同一亚型 ,为HTN型病毒中的H4亚型
Objective To isolate the Hantaan virus in Liaoning Province and genotype the isolate (SY 13) in order to find out the genotypes and genotypes of the Hantaan virus in Liaoning Province. Methods Hantavirus was isolated by tissue culture method. The G1, G2 and S fragments of M segment of SY 13 were amplified by reverse transcription-polymerase chain reaction (RT PCR). The sequence of DNA fragment was amplified by DNAStar, Clustalx and Phylip, The sequences of the gene fragments were analyzed for homology with the sequences downloaded from GenBank and the phylogenetic tree was constructed. Results By comparing and analyzing the three gene fragments, the homology between SY 13 and HTN was high (79.3% -97.0%). According to the analysis of the G2 region of SYS, the homology between SY 13 and BAO 10, BAO 14, JIANG 13, BAO 9 and Q 33 were in the same branch with homology of 95.0% -97.0%. Conclusions The Hantaan virus SY 13 strain in Liaoning Province belongs to the same subtype as the Heilongjiang isolate, which belongs to the H4 subtype of HTN virus