安徽省汉坦病毒的基因序列分析

Gene-analysis of hantavirus in Anhui province

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 learn the genotype and characteristic of hantavirus （ HV }, and provide basis for the prevention and treatment. Methods Positive lung tissues of mice and sera of acute HFRS patients were inoculated in Vera cell line. Total RNA was extracted from the culture, and RT - PCR was applied to amplify the S gene of HV. The full - length se- quence of S gene was obtained by sequencing and assembly. The sequencings were analysed by DNAStar and MEG4.1 to construct the phylogenetic tree, so that it could be used to identify the genotype of HV. Results The immuno fluorescent antibody assay （IFA） showed 21 positive results in 513 lung tissues of mice and 6 strains of virus isolated from the positive tissues ; while 1 strain virus was isolated from 4 sera of acute patients. Compared with type HTN, SEO, SNV strain of HV, S gene of three strains of virus showed that two were type HTN and one was SEO by sequencing and analysis. The analysis of all the strains turned out 98.4% homology with standard strains. Conclusion In conclusion the genotype of HV preva- lent in Anhui province are type HTN and SEO, genetic variants has not been found.