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目的对北京协和医院分离的SARS冠状病毒(SARS-CoV)PUMC01株的基因组序列进行变异及系统发育分析。方法利用随机引物法构建SARS-CoV-PUMC01分离株的cDNA文库,随机挑选质粒克隆进行大规模测序,测序反应经过组装后获得全基因组序列(GenbankAccessionNo.,AY350750),对该序列与SARS-CoV的参考序列相比进行系统发育及变异分析。结果与参考序列(SARS-CoV-Tor2及SARS-CoV-Urbani)相比较,在SARS-CoV-PUMC01上共发现10个变异位点;与其他的17株SARS-CoV进行系统发育分析后发现,18株SARS-CoV分为两类,两类之间及每一类的各株病毒间具有不同的分化时间。结论了解不同地区SARS-CoV之间的系统发育关系,为系统了解不同SARS-CoV分离株之间的临床关系以及SARS-CoV的传播链提供了进化上的证据。
Objective To analyze the variation of the genome sequence of SARS-CoV PUMC01 isolated from Peking Union Medical College Hospital and its phylogenetic analysis. Methods The cDNA library of SARS-CoV-PUMC01 was constructed by random primer method. The plasmid clones were randomly selected for large-scale sequencing. After sequencing, the complete genome sequence was obtained (GenBank Accession No. AY350750) Phylogenetic and variation analysis compared to the reference sequence. Results Compared with the reference sequence (SARS-CoV-Tor2 and SARS-CoV-Urbani), 10 variation sites were found on SARS-CoV-PUMC01. After phylogenetic analysis with other 17 SARS-CoVs, Eighteen isolates of SARS-CoV fall into two categories, with different differentiation times between strains of each type and between strains. Conclusion Understanding the phylogenetic relationships among SARS-CoV in different regions provides evidence for systematic understanding of the clinical relationship between different SARS-CoV isolates and the transmission chain of SARS-CoV.