一种新型胶体金免疫层析双抗体夹心法新型冠状病毒抗原检测试剂的研制  

Development of a novel gold immunochromatographic double-antibody sandwich assay for detection of SARS-CoV-2 antigen

作　　者：姚栩[1,2] 陈智伟 张彩云[1] 谢迺鸿[1] YAO Xu;CHEN Zhiwei;ZHANG Caiyun;XIE Naihong(Fuzhou Center for Disease Control and Prevention,Fuzhou,Fujian 350005,China;不详)

机构地区：[1]福州市疾病预防控制中心,福建福州350005 [2]福建医科大学福州市疾病预防控制中心教学(实习)基地,福建福州350005

出　　处：《中国病毒病杂志》2023年第2期115-119,共5页Chinese Journal of Viral Diseases

基　　金：福州市科技计划项目(2021-XG-021)。

摘　　要：目的 采用胶体金免疫层析技术进行新型冠状病毒(severe acute respiratory syndrome coronavirus 2,SARS-CoV-2)抗原检测试剂的研制,并对制备试剂的主要性能指标进行评价。方法 采用碳酸钾、大颗粒胶体金、SARS-CoV-2抗体制备胶体金抗体标记物,筛选不同浓度SARS-CoV-2抗体进行结合垫制备,选择不同浓度SARS-CoV-2抗体包被工作液进行包被膜的制备,硝酸纤维素膜上分别包被SARS-CoV-2抗体和羊抗鼠IgG多抗作为检测线和质控线,按工艺要求进行组装生产。对研制的抗原检测试剂进行最低检出限、交叉反应性、加速稳定性试验及临床性能评估。结果 试剂检测SARS-CoV-2灭活病毒的最低检出限为3.3×10^(2) TCID_(50)/ml(TCID_(50)为半数组织培养感染剂量);检测10种常见病原体(人冠状病毒、副流感病毒、流感病毒、呼吸道合胞病毒、腺病毒、鼻病毒、肠道病毒、溶血性链球菌、金黄色葡萄球菌、大肠埃希菌)不同浓度样本均无交叉反应;试剂经37℃28 d高温热加速实验,试剂性质稳定;临床性能评估,与核酸检测相比灵敏度为92.00%,特异性为100.00%,总符合率为98.67%;一致性检验Kappa值为0.939,P<0.01。结论 研制的抗原检测试剂检测灵敏度和特异性良好,检测时间短、方法简单、操作便携,可作为新型冠状病毒的大规模筛查一种快速检测方法。Objective To develop a novel gold immunochromatographic double antibody sandwich assay for the detection of SARS-CoV-2 antigen, and to evaluate the performance of major reagents. Methods Potassium carbonate, large colloidal gold and SARS-CoV-2 antibody were used to prepare colloidal gold antibody markers, SARS-CoV-2 antibody concentration was optimized to prepare the binding pad, SARS-CoV-2 antibody and goat anti-mouse IgG were coated on nitrocellulose membrane as detection line and quality control line, according to the process requirements to assembly the assay. The minimum detection limit, cross-reactivity, accelerated stability test and clinical evaluation of the antigen detection reagent were determined. Results The minimum detection limit of SARS-CoV-2 inactivated virus was 3. 3×10^(2) TCID_(50)/ml, and no cross-reaction was found in the samples containing 10 common pathogens. The results of 37 ℃ high temperature accelerated test for 28 d showed high stability of the reagent. The sensitivity, specificity and total coincidence rate were 92. 00%, 100. 00% and 98. 67% and the Kappa value of concordance test was 0. 939, P<0. 01. Conclusion The developed antigen detection assay has high sensitivity and specificity, which is also simple to operate in a short time. It can be used as a rapid detection method for large-scale screening of novel coronavirus.

关 键 词：新型冠状病毒 抗原 胶体金 免疫层析 性能评估 

分 类 号：R446.6[医药卫生—诊断学]