新型冠状病毒IgM-IgG抗体检测试剂盒的制备及对15例患者临床应用初试

目的 开发一种快速检测SARS-CoV-2病毒的IgM-IgG抗体的胶体金免疫层析检测试剂盒,并优化研发及应用策略,分析IgM-IgG抗体联合检测在新冠病毒检测中的临床应用价值。 方法 表达纯化SARS-CoV-2病毒S蛋白的RBD和NTD结构域,通过胶体金制备工艺包被抗原及多克隆抗体,制备IgM-IgG抗体检测试剂盒;收集15例阳性患者的血清进行抗体检测分析,计算试剂盒检测的阳性率,分析SARS-CoV-2抗体IgM-IgG对病毒响应的过程。 结果 在15例核酸RT-PCR检测阳性的患者血清样本中,IgM-IgG联合抗体检测的阳性率为73.33%,其中IgM阳性占总标本数的53.33%,IgG阳性占总标本数的60.00%;5例境外人员中有3例为抗体阳性;对患者的流行病学分析发现,病毒感染1周以上的患者共7例,抗体检测全部为阳性;抗体检测试剂盒的灵敏度73.33%,特异性95.00%。 结论 IgM-IgG联合抗体检测法具有快速、方便、易操作等特点,可以作为核酸检测的辅助手段对疑似患者(尤其是境外人员及无症状患者)进行初筛;且IgM-IgG的检测结果可以作为推测患者病毒感染过程的依据;快速检测IgM-IgG抗体将为COVID-19疾病的诊断和治疗提供帮助。

Development and clinical value evaluation of IgM-IgG antibody detection kit for SARS-CoV-2 infection in 15 cases

Objective To develop a colloidal gold kit for rapid detection of IgM-IgG antibodies of SARS-CoV-2, optimize the development and application strategy, and investigate the diagnostic value of SARS-CoV-2 IgM-IgG antibodies by detecting serum of clinically confirmed patients. Methods The RBD and NTD domains of S protein from SARS-CoV-2 virus were expressed and purified to prepare IgM-IgG antibody colloidal gold detection kit by coating antigen and polyclonal antibody with preparation technique of colloidal gold, then the clinical efficacy was validated. The blood samples collected from 15 PCR confirmed COVID-19 patients were detected and the positive rate was calculated to analyze the response progress of SARS-CoV-2 antibody IgM-IgG on virus. Results The overall positive rate of IgM-IgG antibody assay test was 73.33%, which was higher that single IgM(positive rate: 53.33%)or IgG(positive rate: 60.00%)test. Three cases were positive out of five aliens. The epidemiology analyses showed that 7 positive cases had infected with SARS-CoV-2 virus for more than one week. The sensitivity of the antibody detection kit was 73.33% and the specificity was 95.00%. Conclusion The IgM-IgG combined test method is rapid, convenient and easy to operate, and can be used to carry on the primary screening for the suspected patients, especially the aliens and asymptomatic patients. On the other hand, the IgM-IgG detection result can be used as the basis to infer the virus-infected progress. The rapid dection results of IgM-IgG antibody may be helpful for the diagnosis and treatment on COVID-19.