摘要
目的调查青岛地区肾综合征出血热病毒的流行情况,研究病毒在该地区的分子流行病学特征及基因分型。方法采集64例肾综合征出血热(HFRS)急性期患者血清标本,提取血清中病毒RNA作为模板。根据GenBank中汉坦病毒基因序列,设计HTN型和SEO型的通用外套引物,同时分别设计HTN型特异性引物和SEO型的特异性引物作为内套引物。采用逆转录聚合酶链反应(RT-PCR)扩增汉坦病毒基因组M片段G1区基因序列并测序。测序结果利用DNA Star软件进行核苷酸序列分析。结果在64例标本中用HTN型特异性引物扩增阳性6例,占9%;用SEO型特异性引物扩增阳性25例,占39%;总阳性率为48%。总体来看,青岛地区流行的汉坦病毒以SEO型为主。SEO型汉坦病毒间核苷酸序列的差异相对较小,在核苷酸序列水平其基因的离散度为0.3%～8.9%。HTN型汉坦病毒的变异率较高,其基因的离散度为2.6%～11.2%。结论青岛地区汉坦病毒的流行以SEO型为主,HTN型并存,其中HTN型主要发生在胶南市。
Objective To research the epidemic circumstance and investigate the molecular epidemiological characteristics and genotyping of hantavirus in Qingdao area of Shandong Province.Methods The 64 serum samples were collected from patients of hemorrhagic fever with renal syndrome（HFRS）,and the virus RNA was extracted as pattern.According to the hantavirus gene sequence of GenBank,HTN and SEO universal primers were designed as outer primers,and HTN and SEO specific primers as inner primers.G1 gene sequence of M fragment from hantavirus genome was amplified by reverse transcription polymerase chain reaction（RT-PCR） and analyzed.DNA Star software was used to analyze nucleic acid sequence.Results Six（9%） cases were positive with HTN specific primer,and 25（39%） cases were positive with SEO specific primer.The total positive rate was 48%.SEO type was prevalent in hantavirus epidemiology in Qingdao.The variation of the nucleotide sequences among SEO type（nucleotide sequence divergence ranging from 0.3%-8.9%） was lower than that among HTN type（nucleotide sequence divergence ranging from 2.6%-11.2%）.Conclusions Most isolates of hantavirus are SEO type,while HTN type exists also in Qingdao.Most HTN isolates are detected at Jiaonan area.
出处
《检验医学》
CAS
北大核心
2011年第8期523-526,共4页
Laboratory Medicine
