基于量子点标记技术的免疫层析法检测新型冠状病毒N蛋白IgG抗体研究  

Study of Fluorescence Immunochromatography Based on Quantum Dots for the Detection of SARS-CoV-2 anti-N Protein IgG Antibody

作　　者：于仁峰 邹小仓 邹大阳 李琳昊 王可慧 贺晓明 徐雅晴 秦日辉 莫冬冬 段佳慧 余涛 刘威 郭金鹏[1,2] YU Renfeng;ZOU Xiaocang;ZOU Dayang;LI Linhao;WANG Kehui;HE Xiaoming;XU Yaqing;QIN Rihui;MO Dongdong;Duan Jiahui;YU Tao;LIU Wei;GUO Jinpeng(The PLA Military Medical ResearchInstitution,Beijing 100850,China;The PLA Center for Disease Control and Prevention,Beijing 100070,China;Beijing Hongjinjiu'an Biotechnology Company,Beijing 102299,China)

机构地区：[1]中国人民解放军军事医学研究院,北京市100850 [2]中国人民解放军疾病预防控制中心,北京市100070 [3]北京弘进久安生物科技有限公司,北京市102299

出　　处：《中国全科医学》2022年第14期1741-1749,共9页Chinese General Practice

摘　　要：背景基于目前新型冠状病毒(SARS-CoV-2)引起的新型冠状病毒肺炎(COVID-19),早诊断、早隔离、早治疗是防控传染的重要方法。建立方便快捷的免疫层析检测技术,对于防控SARS-CoV-2的传播十分必要。目的建立基于量子点标记技术的免疫荧光层析法检测抗SARS-CoV-2 N蛋白IgG抗体的方法,判断被检测者是否感染过SARS-CoV-2或者是否接种过SARS-CoV-2灭活疫苗。方法2020年8月将制备好的鼠抗人二抗和抗N蛋白抗体固定至硝酸纤维素(NC)膜上,分别作为检测线(T)和质控线(C),然后将量子点标记的SARS-CoV-2 N蛋白均匀喷洒在玻璃纤维上,干燥后组装、切割、包装成试纸条。用该试纸条分别检测35例COVID-19患者和50例健康人的血清,以酶联免疫吸附测定法(ELISA)试剂盒初筛结果作为对照,计算量子点荧光免疫层析法的检测特异度和灵敏度,并利用N蛋白抗体标准品检测试纸条灵敏度。结果本研究制备的试纸条特异度为100.00%,灵敏度为94.29%,灵敏性为8.53~17.06 ng/ml抗体浓度。结论采用量子点标记技术检测血清中的抗N蛋白IgG抗体,操作简单、快速,灵敏度及特异度强。Background Based on the current prevalence of Coronavirus Disease 2019(COVID-19),early diagnosis,isolation,and treatment are important methods to prevent and control infectious diseases.The establishment of convenient and efficient immunochromatographic detection techniques is essential for the prevention and control of COVID-19 epidemic.Objective To establish a method for the detection of SARS-CoV-2 anti-N protein IgG antibody by immun of luorescence chromatography method based on quantum dots labeling technology in August,2020.In order to determine whether the detected persons had been infected with COVID-19 or been injected with SARS-CoV-2 inactivated vaccine.Methods The prepared rat anti-human secondary antibody and anti-N protein antibody were immobilized on a Nitrocellulose(NC)membrane as detection line(T)and quality control line(C),respectively.Then the SARS-CoV-2 N protein labeled by quantum dots was evenly sprayed on glass fiber,which was assembled,cut and packaged into test strips after drying.The test strips were used to detect the clinical serum of 35 COVID-19 patients and 50 healthy individuals,the results of the initial screening of the ELISA kit were used as a control to calculate the detection specificity and sensitivity of quantum dots fluorescence immunochromatography.The sensitivity of the test strip was detected by using the N protein antibody standard.Results The specificity and sensitivity of the strip were 100.00%,94.29%,and the susceptibility was 8.53-17.06 ng/ml antibody concentration.Conclusion The detection of anti-N protein IgG antibody in serum by quantum dots labeling is simple,fast,with strong sensitivity and specificity.

关 键 词：新型冠状病毒肺炎 新型冠状病毒 免疫层析法 量子点 IGG抗体 N蛋白 

分 类 号：R563.12[医药卫生—呼吸系统]