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目的探讨辽宁省汉滩型汉坦病毒(HTNV)的基因特征及其分布情况。方法在辽宁省肾综合征出血热(HFRS)主要流行地区收集鼠肺和HFRS患者标本,采用间接免疫荧光法检测鼠肺中HV抗原,以RTPCR方法扩增标本中M和S基因片段的特异性核苷酸序列,将HTNV的阳性产物测序,并进行同源性比对和系统发生分析。结果辽宁省HFRS疫区HTNV主要为黑线姬鼠所携带;从鼠肺标本中扩增出M片段4份,患者标本中扩增出M片段5份,S片段1份;M片段的核苷酸同源性分析表明,辽宁株与Bao14、CJAp267等株同源性最高,为95%～97%,与HTNV原型株76-118同源性为87.4%～89.0%;系统发生分析显示辽宁株均分布于同一支内,与Bao14、CJAp267等株构成一个独立的支系,同属于H4亚型;TL2S基因片段与YaluRiver13核苷酸同源性最高(97.1%),与HTNV原型株76-118同源性为91.2%;推导的S片段氨基酸同源性,TL2与Bao14、CJAp93同源性较高,为93.0%～96.2%,与其他代表株同源性多在74.1%～81.6%;而且基于S片段的系统发生分析提示与M片段的分型结果基本一致,与Bao14、CJAp93等株位于同一分支内,为H4亚型。结论目前辽宁省流行的HTNV主要为H4基因亚型,基因亚型的分布相对比较单一。
Objective To investigate the genetic characteristics and distribution of Hantaan hantavirus (HTNV) in Liaoning Province. Methods Specimens of murine and HFRS were collected from the prevalence areas of hemorrhagic fever with renal syndrome (HFRS) in Liaoning Province. The HV antigen was detected by indirect immunofluorescence and the specificity of M and S gene fragments was amplified by RTPCR Nucleotide sequence, the positive product of HTNV was sequenced, and homology alignment and phylogenetic analysis were performed. Results The HTNV of HFRS in Liaoning province was mainly carried by Apodemus agrarius. Four M fragments were amplified from rat lung samples. Five M fragments and one S fragment were amplified from the lung samples. Nucleotide Acid homology analysis showed that Liaoning strains had the highest homology of 95% ~ 97% with strains Bao14 and CJAp267, and 87.4% ~ 89.0% with HTNV prototype strain 76-118. Phylogenetic analysis showed that Liaoning strain They all belong to the same subfamily as Bao14 and CJAp267, belonging to H4 subtype. The TL2S gene fragment has the highest nucleotide homology (97.1%) with YaluRiver13 and the homology with HTNV prototype strain 76-118 The homology was 91.2%. The amino acid homology of deduced S fragment was high. The homologies between TL2 and Bao14 and CJAp93 were 93.0% ~ 96.2%, and 74.1% ~ 81.6% with other representative strains. The phylogenetic analysis based on S-segment suggested that it was consistent with the typing of M segment, and was located in the same branch with Bao14 and CJAp93, which were H4 subtypes. Conclusions Currently, the prevalence of HTNV in Liaoning Province is mainly H4 genotype, and the distribution of genotypes is relatively simple.