论文部分内容阅读
目的 对杆状病毒表达SARS冠状病毒NP抗原性进行分析与比较。方法 利用SDS PAGE、Western Blot和ELISA证明重组核蛋白(rSN)在昆虫细胞获得高效表达,具有特异免疫反应原性。结果 表达rSN的昆虫细胞裂解、稀释后直接包被ELISA板,来检测SARS CoV康复病人血清特异抗体,敏感性稍高于Vero细胞培养的全病毒(OD分别为2 .8和0 .6 ) ,与健康人血清不发生特异反应(OD值为0 .0 1) ;表达rSN昆虫细胞用于间接免疫荧光,快速检测血清特异抗体反应,同样表现了良好的敏感性和特异性。结论 昆虫细胞表达rSN有望替代SARS CoV全病毒,作为安全、敏感和特异的诊断抗原,应用于ELISA和IFA血清学检测。
Objective To analyze and compare the NP antigenicity of baculovirus expressing SARS coronavirus. Methods SDS PAGE, Western Blot and ELISA demonstrated that the recombinant nucleoprotein (rSN) was highly expressed in insect cells and had specific immunogenicity. Results Insect cells expressing rSN were lysed and directly coated with ELISA plate after dilution to detect serum specific antibodies in SARS-CoV patients with slightly higher sensitivity than those in Vero cells (OD 2.8 and 0.6, respectively) No specific reaction with healthy human serum (OD 0. 01); expression of rSN insect cells for indirect immunofluorescence, rapid detection of serum-specific antibody response, also showed good sensitivity and specificity. Conclusion Insect cell expressing rSN is expected to replace SARS CoV full-length virus as a safe, sensitive and specific diagnostic antigen for ELISA and IFA serological tests.