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目的 从浙江省首发的 3例临床诊断SARS患者样本中分离SARS病毒 ,并进行病毒核酸、免疫学检测和形态学观察。方法 采集 3例SARS患者发病期的含漱液 ,接种Vero、Vero -E6、RD、Hep -2、MK单层细胞 ,观察细胞病变、免疫荧光法检测病毒抗原和RT -PCR法检测病毒核酸。抗原阳性的培养物作电镜形态学观察。结果 从 3例患者的 2份含漱液中分离到 2株SARS病毒 ,并发现Vero和RD两种细胞对SARS病毒敏感 ,在接种病毒后的 3天左右可出现典型的细胞病变 ,两株病毒已在Vero和RD细胞上传代 5代 ;在电镜下观察到SARS病毒颗粒。结论 分离的 2株病毒确定为SARS冠状病毒 ,并能在Vero细胞上稳定传代 ;建立的间接免疫荧光技术可用于SARS实验室诊断及流行病学调查等。
Objective To isolate SARS virus from 3 samples of SARS patients diagnosed in Zhejiang province and to detect the virus nucleic acid, immunology and morphological observation. Methods Ringer’s solution was collected from 3 SARS patients and inoculated with Vero, Vero-E6, RD, Hep-2 and MK monolayer cells. The cytopathic effect was observed. The viral antigens were detected by immunofluorescence and RT-PCR. Antigen positive cultures were observed electron microscopy. Results Two SARS viruses were isolated from two gargle samples of three patients and found that both Vero and RD cells were sensitive to SARS virus. Typical cytopathic effect occurred about three days after the virus was inoculated. Two viruses It has been passaged on Vero and RD cells for 5 generations; SARS virus particles were observed under electron microscope. Conclusion The two isolated viruses were identified as SARS coronavirus and could be stably passaged on Vero cells. The established indirect immunofluorescence assay could be used for SARS laboratory diagnosis and epidemiological investigation.