SARS病毒分离与病原学研究

Isolation and etilogic study of SARS-associated virus

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。

Objectives To isolate SARS-associated virus in samples from three confirmed patients with SARS in Zhejiang province , detect the virus nucleic acid and reveal the immunological and morphologic characteristics.Methods Gargles were collected from these three patients during the onset period, and inoculated to Vero,Vero-E6,RD,Hep-2, or MK monolayer cells. Pathologic changes of cells were observed and virus antigens were detected by immunofluorescence after several days of culture at 37℃.Morphologic obseration was carried out for antigen positive culture by electron microscopy.Results Two strains of SARS-associated virus were successfully isolated from two samples from three patients. Both Vrero and RD cells were sensitive to the virus. Typical pathologic changes of cells were observed on the third day after inoculation.TwoVirus strains have passaged 5 generations in Vero and RD cells lines. Granular viruses were found by eletron microscopy.Conclusions The virus we isolated can be identified with SARS-associated coronavirus and stably passaged in Vero Cells Lines. The established indirect immunofluorescence technology may apply to confirmes diagnosis and epidemiological study of SARS.