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采集急性期病人的咽拭子或漱口液,用Vero、Vero E6、MDCK、Hela、Hep-2等传代细胞,人胚肺二倍体细胞(HEL)和人胚肺(HP)细胞分离培养严重急性呼吸系统综合症(SARS)的病原体。结果用Vero、Vero E6、MDCK和HP细胞从标本中分离到一株病毒。间接免疫荧光试验发现,恢复期病人血清可与所分离的病毒起反应,在胞膜和胞浆中出现翠绿色荧光;中和试验结果表明,恢复期病人血清能中和病毒对细胞的致细胞病变作用:电镜下可观察到冠状病毒样颗粒:RT-PCR法可扩增到冠状病毒特异性基因片段,且其核苷酸序列与国内外发表的SARS冠状病毒(SARS-Cov)相应的基因序列相符,同源性达到100％。从传染性非典型肺炎病人的漱口液中分离到SARS冠状病毒,这种病毒与传染性非典型肺炎密切相关。
Throat swabs or mouthwash were collected from patients in acute phase. The cells were subcultured with passage cells of Vero, Vero E6, MDCK, Hela, Hep-2 and so on. Human embryo lung diploid cells (HEL) and human embryo lung Severe Acute Respiratory Syndrome (SARS) pathogens. Results A virus was isolated from the specimens using Vero, Vero E6, MDCK and HP cells. Indirect immunofluorescence test found that the recovery of the patient’s serum can react with the virus isolated in the membrane and cytoplasm appear emerald green fluorescence; neutralization test results show that the recovery of the patient’s serum can neutralize the virus on the cells caused by cells Lesions: coronavirus-like particles can be observed under electron microscopy: RT-PCR amplified coronavirus-specific gene fragments, and the nucleotide sequence and the published SARS-CoV (SARS-Cov) gene Sequence matches, homology reached 100%. The SARS coronavirus is isolated from the mouthwash of SARS patients and is closely associated with SARS.