汉坦病毒浙江分离株ZT71株的全基因核苷酸序列测定及分析

Complete genome sequence analysis of the Hantavirus ZT71 strain

目的为了获得浙江省汉坦病毒基因组更为详尽的资料，研究汉坦病毒的进化状况及变异程度，为疫苗病毒株的选择使用提供科学依据。方法设计特异性引物，RT—PCR分段扩增ZT71株全长L、M和S片段，PCR产物纯化后克隆于T载体并进行序列测定。然后进行遗传进化分析。结果汉坦病毒ZT71株L、M及S基因分别由6530、3651和1753个核苷酸组成，分别编码2151、1133和429个氨基酸。基因比较分析表明，ZT71株与SEO型汉坦病毒比较其L片段核苷酸同源性为95．5％～99．7％，氨基酸同源性为99．0％～99．3％；M片段核苷酸同源性为84．1％～99．6％，氨基酸同源性为95．3％～99．2％；S片段核苷酸同源性为88．7％～99．5％，氨基酸同源性为96．5％～99．1％；而与其他汉坦病毒同源性则较低。结论测定了ZT71株的全基因核苷酸序列，其L、M和S片段具有和其他汉坦病毒L、M、S片段相似的核苷酸一级结构，但与SEO型更为接近，属于SEO型病毒。

Objective To study the complete genome sequence of Hantavirus ZT71 strain gene isolated in Zhejiang province and explore its evolution. Methods The total RNA was prepared from ZT71 virus infected cells and the RT - PCR products were cloned into T vector , sequenced and analyzed. Results The L, M and S segments of the strain ZT71 genome were 6530, 3651 and 1753 nucleotides in length with a single open reading frame individually encoding 2151, 1133 and 429 amino acids. The sequence analysis of nucleotides showed that the homology of L, M and S segments of strain ZT71 between those of other strains of Seoul virus could reach 95.5% -99.7% , 84.1% -99.6% and 88.7% -99.5%, respectively. The analysis of the deduced amino acids showed the similar result. The source of strain ZT71 could be traced from the analysis of the phylogentic trees of nucleotides and amino acids, and it should belong to Seoul type of Hantavirus which was also verified serologically. Conclusion The nucleotide sequences and deduced amino acid sequences of L, M and S segments of strain ZT71 are similar to that of those of Seoul type of Hantavirus. And Hantaan type virus used to be prevalent primarily in Zhejiang province, and it would be an endemic area of mixed type of Hantavirus since the discoveries of the viruses of Soeul type in recent years.