河北省人感染汉坦病毒G2片段基因分型及序列特征分析

Genotype and sequence analysis on G2 segments of Hantavirus from HFRS patients in Hebei Province

目的了解河北省人感染汉坦病毒的基因型和基因亚型。方法根据76-118株和1122株的G2编码区设计型特异性引物，利用RT-nestedPCR技术对采集的HFRS患者中汉坦病毒抗体阳性的血清标本进行检测及基因分型，从中选取部分标本的扩增产物纯化回收后进行核苷酸序列测定，用DNAStar软件包进行基因分析。结果69份阳性血清标本经RT-nested PCR检测17份阳性，检出率为24．64％，其中≤7d的标本检出率为34．29％，8-14d的检出率为19．23％，≥14d的未能检出。17份阳性标本均为SEO型。对其中9份扩增产物的G2区测序后表明，9份血清标本的同源性为92．0％-100．0％，其中HeB7与其他8份标本的同源性为92．0％。95．0％，属不同亚型，与1122的同源性为97．7％，同属于S1亚型；除HeB7外其余8份标本同源性高达95．7％-100％，同属于S3亚型。9份标本与76-118株的同源性仅为70．3％-72．7％。结论河北省人感染汉坦病毒以SEO型为主，亚型以S3为主，也存在S1亚型。在一定范围内，同型汉坦病毒基因相对保守，具有区域稳定性特征。

Objective To know the genotype and subtype of Hantan virus （HV） which infected persons in Hebei province. Methods According to G2 coding region of 76-118 and R22 strains, specific type primers were designed to detect and identity the types of HV in HFRS patients＇ sera with RT-nested PCR. Nucleotides were assayed from partial products after purification and reclaim. Then, gene analysis was done with DNAStar package. Results 17 out of 69 positive serum specimens were successfully detected by RT-PCR and the detection rate was 24.64%, among which, ≤7days was 34.29%, 8-14 days was 19.23%, and ≥14 days were 0.17 positive specimens were all belonged to SEO. The nucleotide homology of 9 typical specimens was 92.0%-100%. Between HeB7 and other 8 specimens was 92%-95%, and they belonged to different subtypes. When HeB7 compared with R22 strain, it was 97.7%. HeB7 and R22 belonged to S1 subtype. The 8 specimens except HeB7 was 95.7%- 100% and they all belonged to S3 subtype. When compared with 76-118 strain, 9 specimens＇ nucleotide homology was orrly 70.3 %-72.7 %. Conclusion SEO was the major type of HV from HFRS patients in Hebei province, S3 was the major subtype and SI was aLso existed. In a certain area, the HV which belonged to the same type was correspondingly conservative, and had the characteristic of regional stability.