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从浙江省3例SARS患者中收集含漱液标本,经处理后接种Vero、RD、VeroE6和Hep-2细胞进行病毒分离,培养3d后在Vero和RD细胞中可观察到细胞病变。从细胞培养上清中提取病毒核酸,用SARS冠状病毒特异性引物进行RT-PCR,并经测序证实从3份临床样本中分离到2株SARS冠状病毒株。对其中1株病毒的基因组进行了全序列测定并作系统进化树分析显示浙江省SARS冠状病毒株与新加坡2774株和台湾TW1株最为接近。
Gargle samples were collected from 3 SARS patients in Zhejiang Province. After treatment, Vero, RD, VeroE6 and Hep-2 cells were inoculated for virus isolation. Cytopathic effect was observed in Vero and RD cells after 3 days of culture. The virus nucleic acid was extracted from the cell culture supernatant, and SARS-CoV-specific primers were used for RT-PCR. Sequencing confirmed that 2 SARS-CoV isolates were isolated from 3 clinical samples. The whole genome of one strain of the virus was sequenced and phylogenetic tree analysis showed that the SARS coronavirus strain in Zhejiang Province was closest to Singapore 2774 strain and Taiwan TW1 strain.