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目的了解安徽省汉坦病毒(HV)型别及分子特征,为防治提供科学依据。方法将汉坦病毒抗原阳性的鼠肺及急性期病人血清标本接种Vero细胞,分离汉坦病毒。阳性培养物提取病毒总RNA,应用RT-PCR扩增病毒S片段,产物经测序拼接获得S片段全基因序列,用DNAStar、MEGA 4.1等生物软件进行核苷酸序列分析,构建基因序列系统进化树,确定病毒型别。结果免疫荧光检测,513鼠肺标本21份阳性,分离6株病毒;4份急性期病人血清分离1株病毒。选择3株病毒进行S片段基因测序,测定序列与汉坦病毒HTN、SEO、SNV型毒株比较,表明两株为HTN型,一株为SEO型,序列相似性达98.4%以上。结论安徽省流行的HV为HTN型和SEO型,以HTN型为主,未发生较大变异。
Objective To understand the type and molecular characteristics of Hantaviruses (HV) in Anhui Province and provide a scientific basis for prevention and treatment. Methods The Hantaan virus positive rat lung and the acute phase serum samples were inoculated into Vero cells and the Hantaan virus was isolated. Positive cultures were used to amplify the total RNA of the virus. The full-length S-segment gene was amplified by RT-PCR and sequenced. The nucleotide sequences were analyzed by using biological softwares such as DNAStar and MEGA 4.1 to construct the phylogenetic tree , Determine the virus type. Results Immunofluorescence assay showed that 21 of 513 rat lung specimens were positive, and 6 strains of viruses were isolated. Four strains of virus were isolated from serum of patients in acute phase. The results showed that the two strains were HTN type, one was SEO type, and the sequence similarity was 98.4%. Conclusion The prevalence of HV in Anhui province is HTN type and SEO type, with HTN type as the main type, with no major variation.