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应用RT -PCR扩增了皖南山区分离株AH0 9的M和S片段全基因 ,克隆于T载体 ,纯化后测定序列。结果AH0 9株M片段的全基因序列共 36 2 5个核苷酸 ,编码 1135个氨基酸 ;S片段的全基因序列共 172 4个核苷酸 ,编码 430个氨基酸。M和S片段全基因核苷酸和氨基酸与汉坦病毒各型毒株的代表株和HTN型毒株的同源性比较表明 ,AH0 9株分枝与HTN型接近 ,与其它各型病毒则相距较远 ,故确定为HTN型毒株。但AH0 9株与HTN型毒株的M和S片段全基因序列有差异 ,其差异分别高达 2 3 6 %和 2 0 4%。经种系发生分析 ,AH0 9株是迄今为止所发现的HTN型病毒中差异最大的新基因亚型病毒株。AH0 9株病毒M片段的氨基酸与HTN型相差 13 5 %至14 8% ,而S片段仅相差 7%～ 8 1% ,说明AH0 9毒株的变异主要发生在M片段。而ORF和 3′端的NCR区核苷酸序列分析比较说明 ,病毒的变异更主要集中在该片段的 3′端的NCR区
The whole gene of M and S fragments of AH0 9 isolate from mountain area in South Anhui was amplified by RT-PCR and cloned into T vector. After purification, the sequence was determined. Results The whole genome sequence of AH0 9 M fragments was 3625 nucleotides in length and encoded 1135 amino acids. The full-length sequence of S fragment contained 172 4 nucleotides and encoded 430 amino acids. Comparison of nucleotide and amino acid sequences of M and S fragments with those of Hantavirus strains and HTN strains showed that the branches of AH0 9 were close to those of HTN, Far apart, it was identified as HTN strains. However, there were differences in the sequences of M and S fragments between AH0 9 strains and HTN strains, with the difference being as high as 236% and 20% respectively. According to the phylogenetic analysis, AH0 9 strain is the most significant new genotype of HTN virus discovered to date. The amino acids of the 9 strains of AH0 were 13 5% to 14 8% different from those of HTN, but only 7% ~ 81% of the S fragments, indicating that the variation of AH0 9 occurred mainly in the M fragment. However, nucleotide sequence comparison of ORF and 3 ’NCR region showed that the variation of the virus was mainly concentrated in the NCR region at the 3’ end of the fragment