SARS病毒不同基因区抗原与SARS患者血清反应性的研究

目的研制非典型肺炎病毒(SARS病毒)不同基因区抗原,探讨SARS病毒感染机体不同基因区抗体免疫应答。方法根据目前已知SARS病毒的基因组序列及其编码的蛋白质,运用计算机抗原表位预测技术,筛选4种SARS病毒主要抗原表位。合成抗原表位基因,克隆到表达载体pBVILl,在E．coli中表达S蛋白和N-蛋白,E-蛋白和M-蛋白为合成肽。结果用SARS病毒编码的S(Spike)蛋白、M-蛋白、N-蛋白和E-蛋白分别与46份SARS患者恢复血清反应,检测血清中IgG抗体,结果S蛋白、N-蛋白、E-蛋白和M-蛋白检出率分别为71．7％(33／46),89．13％(41／46),30．43％(14／46),52．17％(24／46)。结论感染SARS病毒病人血清中S蛋白、M-蛋白、N-蛋白和E-蛋白的免疫应答有显著差异。

Reactivity of Antigens Derived from Different Structural Genes of SARS-CoV to Convalescent Sera

Objective To study the reactivity of recombinant and synthetic antigens derived from different structural genes of SARS-Coronavirus toward convalescent sera of SARS patients (as a probe into the humoral immune response induced by the virus).Methods Multi-epitope chimeric recombinant SARS-CoV "Spike" protein and "Nucleocapsid" protein antigens and synthetic E & M peptide antigens were constructed using epitope prediction software to scan the published SARS-CoV genomic sequence.Purified antigens were used to coat microplates and presence of corresponding antibodies in convalescent sera was detected using an indirect ELISA format.Results The number of positive reactive sera over the number of convalescent sera tested using the recombinant S and N antigen of SARS-CoV and E and M peptide antigens were as follows:"S" 33/46(71.7%),"N"41/46(89.13%),"E"14/46(30.43%) and"M"24/46(52.17%).Conclusion There exist quite marked differences among antibody responses in SARS convalescent sera when tested with recombinant or synthetic antigens derived from different structural genes.