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  

Clinical application study on SARS-CoV spike protein antigenic specificity

作　　者：段朝晖[1] 罗晓红[1] 刘然义[2] 邵静[1] 黄嘉凌[2] 朱振宇[3] 黄健[1] 黄文林[2] 

机构地区：[1]中山大学附属第二医院检验科,广州510120 [2]中山大学附属肿瘤研究所 [3]中山大学中山医学院生化教研室

出　　处：《中华检验医学杂志》2005年第9期902-904,共3页Chinese Journal of Laboratory Medicine

基　　金："国家973"重大基础研究发展计划基金资助项目(2003CB514107)

摘　　要：目的探讨严重急性呼吸道综合征(SARS)患者的SARS冠状病毒(SARSCoV)抗体产生情况以及SARSCoVSpike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SARSCoVIgM总阳性率为91.6%(87/95),抗SARSCoVIgG总阳性率为97.9%(93/95);高暴露人群组和正常对照组的抗SARSCoVIgM和抗SARSCoVIgG阳性率为0%。经过携带Spikeprotein基因的重组病毒AdSn感染COS1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CNE2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SARSCoV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SARSCoV的Spike基因所表达的Spike蛋白。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 Spikeprotein 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.

关 键 词：严重急性呼吸综合征 抗原 抗体 印迹法 蛋白质 严重急性呼吸道综合征(SARS) SARS冠状病毒 抗原特异性 临床应用研究 E蛋白 SARS患者 

分 类 号：R446.6[医药卫生—诊断学]