河南省部分区域肾综合征出血热患者汉坦病毒的分子特征

Molecular characteristics of hantavirus isolated from patients with hemorrhagic fever with renal syndrome(HFRS)in several regions of Henan Province

目的分析河南省部分区域肾综合征出血热患者汉坦病毒的分子特征。方法采集2017-2018年就诊于河南省郑州市第六人民医院临床诊断为肾综合征出血热患者血标本32例,采用半巢式PCR筛选汉坦病毒阳性样本,然后再对阳性样本中病毒S片段和M部分片段基因进行扩增和测序,并对得到的S片段基因序列进行组装。所得序列与已分离到的国内其他省份相对应的基因序列进行同源分析并构建系统发生树。结果收集的32份血标本中,16份血标本扩增出目的片段,序列分析表明均为汉滩病毒(HTNV)。系统发生分析显示16株病毒株S片段和M部分片段聚类分支关系基本一致。其中HN2018/138病毒株与其余15株S片段和M部分片段核苷酸序列的同源性分别为91.4%~92.1%和83.2%~84.4%,差异较大,其余15株亲缘关系较近。结论本研究所统计的河南省部分区域内肾综合征出血热患者以HTNV为主导基因型,且其表现出明显的地域聚集现象,并存在基因差异较大的变异株或省外输入病毒株的可能性。

To investigate the genotype and genetic characteristics of hantavirus(HV)in regions of Henan Province,we collected 32 blood specimens from patients with hemorrhagic fever with renal syndrome(HFRS)admitted to the Sixth People’s Hospital of Zhengzhou between 2017 and 2018.Hantavirus RNA was reverse transcribed,and regions bearing the S gene sequence and partial M gene sequence were amplified with hemi-nested PCR,sequenced,and manually assembled.Nucleotide sequence similarity among the newly determined sequences along with sequences from other provinces was analyzed,and a phylogenetic tree was constructed with the neighbor-joining method.Of the 32 serum samples,the HV genome fragment was successfully amplified and sequenced in 16 samples,and sequence analysis indicated that all isolates belonged to Hantaan virus(HTNV).The constructed NJ trees based on S and partial M sequences showed similar topological structures.An HN2018/138 strain sharing 91.4%92.1%and 83.2%84.4%similarity with the other 15 strains at the S segment and partial M segment sequences,respectively,formed a single branch.HTNV,the main cause of HFRS in the studied regions of Henan province,showed clear geographic clustering and high genetic variation,possibly due to mutations or imported cases.