新型冠状病毒抗原检测双抗体夹心ELISA方法的建立

Establishment of double antibody sandwich ELISA for detection of 2019-nCoV antigen

目的探索即时、快速、准确的新型冠状病毒(2019-nCoV)抗原检测方法,提高新型冠状病毒感染者检出率,并建立2019-nCoV的S蛋白特异性检测方法。方法本研究利用ELISA方法对前期研发的2019-nCoV基因工程单克隆抗体(mAb)进行配对检测,从中挑选出最优检测抗体组合,建立了新型冠状病毒抗原检测双抗体夹心ELISA方法,并对检测方法进行条件优化。结果本实验室前期筛选出的12株2019-nCoV基因工程单克隆抗体对2019-nCoV S蛋白具有良好的结合活性,是抗原检测候选抗体。本研究所建立的新型冠状病毒抗原检测双抗体夹心ELISA方法可快速有效的检测2019-nCoV S蛋白,灵敏度较高,对S蛋白检出限小于1 ng/ml。结论本研究所建立的抗原检测方法可特异性检测2019-nCoV的S蛋白,为COVID-19感染的早期、敏感、特异诊断提供了有意义的参考。

Objective To explore an early,rapid and accurate detection method of severe acute respiratory syndrome coronavirus 2(2019-nCoV)antigen and improve the detection rate of patients with 2019-nCoV.Methods We detected the characteristics of monoclonal antibodies(mAbs)against 2019-nCoV generated previously and established a double antibody sandwich ELISA against S protein of the virus.Results All 12 mAbs against 2019-nCoV screened in our previous study were able to recognize purified virion and S protein.They were good candidates for detecting antibody to 2019-nCoV antigen.A double antibody sandwich ELISA established in this study could rapidly and effectively detect 2019-nCoV S protein with high sensitivity.The detection limit of 2019-nCoV S protein was lower than 1 ng/ml.Conclusions This method could specifically detect 2019-nCoV antigen,and provided a meaningful reference for early,sensitive and specific diagnosis of COVID-19 infection.