目的：探讨PCR-测序法在啮齿类动物汉坦病毒临床检测中的应用价值。方法以Genbank 7种血清亚型24株汉坦病毒代表性毒株为基础，从病毒基因S片段序列设计引物，采用邻位相连法进行系统进化分析，以该方法对浙江省近年从野生啮齿类动物中临床分离的汉坦病毒毒株进行分型鉴定。结果所建系统发育树将所分析的毒株分为5个区域，引起HFRS的四种血清型具有较稳定的拓扑结构，且能与引起HPS的血清型进行区分。11株实验毒株进行PCR扩增和测序，结果表明该引物具有高度的敏感性和特异性，其中9株浙江省分离的血清型未知毒株的系统发育分析发现其包含引起HFRS的3株HTN和1株SEO血清型，其他5株属两种未知血清型。讨论所建立的PCR-测序法具有用于临床检测汉坦病毒的价值。%Objective To evaluate the application value of PCR-sequencing in clinical detection of hantavirus in rodents .Methods Based on 7 subtypes and 24 strains of representative hantavirus strains downloaded from Genbank , the virus S gene fragments were used for primer design and neighbor joining method was applied for phylogenetic analysis . Thereafter, we identified hantavirus strains isolated from wild rodents in recent years in Zhejiang Province by this method . Results The 24 analyzed strains were divided into 5 regions in the phylogenetic tree .Four of them with topology structure were more stable .Eleven strains of the virus were amplified by PCR and sequenced , and the results showed that the prim-ers were with high sensitivity and specificity .Three HTN strains and 1 strain of serotype SEO were distinguished from 9 strains of unknown strains isolated in Zhejiang Province .We also found that 5 strains of hantavirus belonging to two un-known serotypes .Discussion Our results suggest that the PCR-sequencing method proposed in this study can be used for clinical detection of hantavirus .
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