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①目的 调查山东省青岛地区 2 0 0 0～ 2 0 0 3年汉坦病毒的流行情况 ,研究病毒在该地区的分子流行病学特征。②方法 采集肾综合征出血热 (HFRS)急性期病人血清标本 6 4份 ,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 ,RT nest 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DNAStar软件进行核苷酸序列分析。③结果 在 6 4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 2 5例 ,占 39% ;总阳性率为 4 8%。总体来看青岛地区流行的汉坦毒株以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 %。④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 investigate the prevalence of Hantavirus from 2000 to 2003 in Qingdao, Shandong Province, and to study the molecular epidemiological characteristics of the virus in the area. Methods Sixty-four serum samples of patients with hemorrhagic fever with renal syndrome (HFRS) were collected. The viral RNA was extracted from the serum as a template. According to the Hantavirus gene sequence in GenBank, HTN-type and SEO-type universal coat primers were designed respectively. HTN-specific primers and SEO-specific primers were designed as inner primers. RT nest PCR was used to amplify the G1 region of M segment of Hantavirus genome and sequenced. Sequencing Results Nucleotide sequence analysis was performed using DNAStar software. ③ Results In 6 4 specimens, 6 cases were positive with HTN-specific primers, accounting for 9%. 25 cases were positive with SEO-specific primers, accounting for 39%. The total positive rate was 48%. Overall, the prevalence of hantavirus strains in Qingdao mainly SEO type. The difference between the SEO-type Hantavirus nucleotide sequences is relatively small, with the gene divergence of 0.3% -8.9% at the nucleotide sequence level. HTN type Hantavirus mutation rate is higher, the gene dispersion degree of 2.6% ~ 11.2%. ④ Conclusion The prevalence of Hantavirus in Qingdao is mainly SEO type, HTN type co-exist, of which HTN type occurs mainly in Jiaonan