严重急性呼吸道综合征冠状病毒Spike蛋白抗原特异性的临床应用研究

目的 探讨严重急性呼吸道综合征（SARS）患者的SARS冠状病毒（SARS—CoV）抗体产生情况以及SARS-CoVSpike蛋白抗原的特异性。方法 运用酶联免疫法（ELISA）和蛋白质印迹法（WesternBlotting）检测95例SARS患者体内抗体的产生情况和验证病毒Spike蛋白抗原的特异性。结果 95例SARS患者血清抗体结果中，大部分患者出现抗体阳性，抗SARS-CoVIgM总阳性率为91．6％（87／95），抗SARS-CoVIgG总阳性率为97．9％（93／95）；高暴露人群组和正常对照组的抗SARS-CoVIgM和抗SARS-CoVIgG阳性率为0％。经过携带Spike-protein基因的重组病毒Ad-sn感染COS-1细胞后表达的s蛋白抗原，使用SARS患者抗血清能够检测出特异性蛋白条带；重组病毒感染CNE-2细胞后，表达的S蛋白抗原，能够通过不同患者抗血清同时检测到相应的特异性抗原蛋白。结论 SARS患者感染SARS-CoV后，体内可产生相应抗体；利用SARS患者所产生特异性抗血清，可以检测出克隆SARS-CoV的Spike基因所表达的Spike蛋白。

Clinical application study on SARS-CoV spike protein antigenic specificity

Objective Study to how the antibody against SARS associated coronavirus (SARS-CoV) develop in patients with severe acute respiratory syndrome (SARS), and the specificity of SARS-CoV Spike protein antigen.Method Enzyme-linked Immuno-sorbent Assay (ELISA) and western blotting were used to detect the antibody produced by the 95 patients with SARS and test the specificity of virus spike protein antigen.Result Most cases of the 95 patients were positive in serum antibody detecting; total positive rate of anti-SARS-CoV IgG was 91.6% (87/95). Total positive rate of anti-SARS-CoV IgM was 97.9% (93/95). While positive rate of both anti-SARS-CoV IgG and anti-SARS-CoV IgM were 0% in either of dangerous group and control. After being infected by recombinant virus Ad-Sn carrying Spike-protein gene, COS-1 cell expressed protein antigen. The anti-serum of patients with SARS could detect the specific protein band of this protein. The protein,expressed by CNE-2 cell (the latter was infected by recombinant virus), could be detected relevant specific antigenic protein through different serums of different patients.Conclusion Relevant antibody was produced in patients with SARS after they were infected by SARS-CoV. Spike protein, expressed by spike gene (the latter was cloned from SARS-CoV), could be detected by specific anti-serum of SARS patients through Western Blotting.