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目的 :从患者尸解肺组织中直接扩增冠状病毒部分聚合酶基因序列 ,为重症急性呼吸综合征 (非典型肺炎 )的病原确认提供依据。方法 :采用RT_PCR方法 ,从非典型肺炎病例标本中进行冠状病毒聚合酶基因的扩增与序列测定。采用ClustalX1.8软件进行系统发生树分析。结果与结论 :从非典型肺炎病例尸解肺组织、传代发病小鼠肺组织及分离物感染的细胞培养物中 ,可分别扩增出冠状病毒的DNA片段。测序结果显示其核苷酸序列与目前已知冠状病毒的同源性在 6 0 %左右 ,而与加拿大和美国最近报道的新型冠状病毒的序列是一致的
OBJECTIVE: To directly amplify the sequence of coronavirus polymerase gene from lung tissue of patients with autopsy, and to provide evidence for the pathogenesis of severe acute respiratory syndrome (SARS). Methods: RT-PCR was used to amplify and sequence the coronavirus polymerase gene from SARS cases. Phylogenetic tree analysis was performed using ClustalX 1.8 software. RESULTS AND CONCLUSION: The DNA fragments of coronavirus can be amplified separately from the cell cultures infected with SARS, lung tissues and isolates infected with atypical pneumonia. Sequencing results showed that the nucleotide sequence was about 60% homologous to the known coronavirus at present, but consistent with the recently reported novel coronavirus sequences in Canada and the United States