利用荧光免疫层析技术检测新冠病毒IgG抗体  

Fluorescence immunochromatography for the detection of SARS-CoV-2 IgG antibody

作　　者：聂聪[1] 刘艳丽[2] 侯宝翠 王静[1] 周永运[3] 孙薇 杨宇[1] NIE Cong;LIU Yan-li;HOU Bao-cui;WANG Jing;ZHOU Yong-yun;SUN Wei;YANG Yu(Institute of Health Inspection and Quarantine,Chinese Academy of Inspection and Quarantine,Beijing 100176,China;不详)

机构地区：[1]中国检验检疫科学研究院卫检所,北京100176 [2]湖北医药学院附属东风总医院 [3]中国合格评定国家认可中心 [4]广州海关

出　　处：《中国国境卫生检疫杂志》2021年第3期156-161,共6页Chinese Journal of Frontier Health and Quarantine

基　　金：国家重点研发计划项目(2018YFF0214900);十堰市级新冠应急项目(20Y61)。

摘　　要：目的建立新冠病毒IgG抗体的快速检测方法。方法制备并纯化重组新冠病毒N蛋白,将获得的重组N蛋白免疫家兔,获得多克隆抗体。将鼠抗人IgG单克隆抗体和兔抗新冠病毒N蛋白多克隆抗体分别喷于硝酸纤维素膜的检测线和控制线,用重组N蛋白标记荧光纳米颗粒作为标记抗原,建立荧光免疫层析方法。结果重组新冠病毒N蛋白经表达纯化后,纯度达到90%以上。用新冠病毒IgG抗体标准品测得建立的荧光层析方法灵敏度为1∶5 000;与甲型流感病毒(H1N1、H3N2、H5N1、H7N9)、乙型流感病毒、呼吸道合胞病毒和腺病毒IgG抗体无交叉反应;批内、批间变异系数均低于10%。结论建立的新冠病毒IgG抗体检测方法具有较好的灵敏性和特异性,可用于新冠病毒抗体的快速筛查。Objective To develop an immunochromatography method for the detection of IgG antibody against SARSCoV-2. Methods The recombinant N protein of SARS-CoV-2 was prepared and purified. The polyclonal antibody was obtained by immunizing rabbits with recombinant N protein. The mouse anti-human IgG monoclonal antibody and rabbit anti-recombinant N protein polyclonal antibody were sprayed on the detection line and control line of the nitrocellulose membrane,respectively. The fluorescent immunochromatography method was established by using fluorescent nanoparticles labeled with recombinant N protein as labeled antigen. Results The recombinant N protein was expressed and purified with a purity of more than 90%. The detection sensitivity of the established fluorescence chromatography method was 1∶5 000 by using standard IgG antibody of SARS-CoV-2. There was no cross-reaction with IgG antibody against influenza A virus(H1N1,H3N2,H5N1,H7N9),influenza B virus,respiratory syncytial virus and adenovirus. The coefficients of variation within and between batches were all less than 10%. Conclusion The established IgG detection method showed good sensitivity and specificity,and can be used as a rapid screening method for SARS-CoV-2 antibody.

关 键 词：新冠病毒 核衣壳蛋白 荧光免疫层析 IGG抗体 

分 类 号：R183.3[医药卫生—流行病学]