基于量子点免疫层析技术的新型冠状病毒中和抗体检测方法建立

Establishment of 2019 novel coronavirus neutralizing antibody detection method based on quantum dotimmunochromatography

目的建立简便、快速、低成本的2019新型冠状病毒(2019 novel coronavirus,2019-nCoV)中和抗体检测方法。方法基于量子点荧光免疫层析原理,建立新冠RBD IgG检测方法,并用新型冠状病毒肺炎(coronavirus disease 2019,COVID-19)患者恢复期血清(N=97)、疫苗免疫后血清(N=82)及健康人血清(N=299)对检测方法性能进行评价。同时用指尖血和外周血配对血样(N=54)对指尖血的适用性进行了评价。结果本研究成功建立了基于量子点荧光免疫层析法2019-nCoV RBD IgG检测方法,该方法检测结果与微量中和试验结果具有较强的相关性(Spearman r>0.73,P<0.0001)和较好的一致性(Kappa=0.93,P<0.01);敏感性和特异性较高,分别为92%和99%。指尖血和外周血检测结果差异无统计学意义(P=0.1026),二者具有较强的相关性(Spearman r=0.9326,P<0.0001);指尖血可用于检测。结论本研究建立的2019-nCoV中和抗体检测方法可为群体免疫状态评估提供的即时、高效,低成本的方法选择,为疫苗群体免疫监测和疫情防控提供技术支撑。

Objective To establish a simple,rapid and low-cost 2019 novel coronavirus(2019-nCoV)neutralizing antibody detection method.Methods The 2019-nCoV RBD specific immunoglobulin G(RBD-IgG)detection method was established based on the principle of quantum dot immunochromatography(QDs),and the detection was evaluated by using of sera from coronavirus disease 2019(COVID-19)convalescent patients(N=97),vaccinated donors(N=82)and healthy donors(N=299).The suitability of fingertip blood was evaluated by matching blood samples with peripheral blood(N=54).Results The 2019-nCoV RBD-IgG detection method based on QDs was successfully established.The detection result of QDSs had strong correlation(Spearman r>0.73,P<0.0001)and good consistency(Kappa=0.93,P<0.01)with the result of micro-neutralization test(MNT).The sensitivity and specificity were 92%and 99%,respectively.There was high correlation(Spearman r=0.9326,P<0.0001)and no significant difference(P=0.1026)between result of fingertip blood and peripheral blood.Fingertip blood can be used as a surrogate sample for testing.Conclusions The 2019-nCoV neutralizing antibody detection method established in this study can provide an immediate,efficient and low-cost method selection for the assessment of herd immunity status,and provide technical support for the herd immunity monitoring of 2019-nCoV vaccinated population and the prevention and control of the epidemic.