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目的;从SARS患者尸检肝、脾、心、肺组织中分离病毒并鉴定及评定中和抗体与SARS病程的关系. 方法:取各组织100g/L匀浆后的上清,接种非洲绿猴肾单层细胞,观察细胞病变;采用RT-RCR及序列分析等技术鉴定病毒并研究其生物学特性;将收集的SARS患者早期和恢复期血清做中和实验. 结果:分别从SARS患者尸检肝、脾、心、肺等组织中分离到致细胞病变的病毒,用各组织的病变细胞提取的RNA为模板分别扩增出部分S区和N区的DNA片段,经DNA序列分析表明所有S区和N区的核苷酸序列均与GenBank上的SARS冠状病毒该区域的序列完全一致,中和实验显示所做10份SARS患者双份血清均有中和抗体效价的4倍升高,轻症患者中和抗体产生时间明显早于重症患者. 结论:从SARS患者尸检肝、脾、心、肺组织中分离到的均为SARS冠状病毒,证实SARS冠状病毒可直接侵害患者的肝、脾、心、肺脏器.所有SARS患者均可产生中和抗体,中和抗体产生时间与疾病进程有一定关系.
Objective To isolate the virus from the liver, spleen, heart and lung tissue of SARS patients and to identify and evaluate the relationship between the neutralizing antibody and the course of SARS.Methods: The supernatant of 100g / Monolayer cells were used to observe the cytopathic effect.The viruses were identified by RT-RCR and sequence analysis and their biological characteristics were studied.The neutralization of early and recovery sera of SARS patients was done.Results: Spleen, heart, lung and other tissues of cytopathic virus isolated from the tissue of the diseased cells RNA extracted as part of the template were amplified DNA fragments of S and N regions, DNA sequence analysis showed that all S and The nucleotide sequence of N region was exactly the same as that of the SARS coronavirus in GenBank. Neutralization experiments showed that the double sera of 10 SARS patients had four times higher neutralizing antibody titers, The time of neutralizing antibody production in patients was significantly earlier than that in severe patients.Conclusion: SARS coronavirus was isolated from the liver, spleen, heart and lung tissues of autopsy patients with SARS and confirmed that SARS coronavirus could directly invade the liver, spleen and heart , Lungs. All SAR S patients can produce neutralizing antibodies, neutralizing antibody production time and disease process have a certain relationship.