新型冠状病毒和甲/乙型流感病毒抗原联合快速检测试剂的研制及性能评价  

Development and performance evaluation of a new rapid detection reagent for novel coronavirus and influenza A/B virus antigen

作　　者：黄涛 钱纯亘 魏道舜 刘仲明 李辉军 汪大明 张赛 HUANG Tao;QIAN Chungen;WEI Daoshun;LIU Zhongming;LI Huijun;WANG Daming;ZHANG Sai(Shenzhen YHLO Biotech Co.,Ltd.,Shenzhen,Guangdong 518116,China;College of Life Science and Technology,Huazhong University of Science and Technology,Wuhan,Hubei 430074,China;General Hospital of Southern Theater Command,Guangzhou,Guangdong 510010,China;Tongji Hospital affiliated to Huazhong University of Science and Technology,Wuhan,Hubei 430030,China;Suzhou Institute of Biomedical Engineering and Technology,Chinese Academy of Sciences,Suzhou,Jiangsu 215163,China)

机构地区：[1]深圳市亚辉龙生物科技股份有限公司,广东深圳518116 [2]华中科技大学生命科学与技术学院,湖北武汉430074 [3]中国人民解放军南部战区总医院,广东广州510010 [4]华中科技大学同济医学院附属同济医院,湖北武汉430030 [5]中国科学院苏州生物医学工程技术研究所,江苏苏州215163

出　　处：《国际检验医学杂志》2023年第S01期65-70,共6页International Journal of Laboratory Medicine

基　　金：深圳市科技创新委员会“新型冠状病毒肺炎疫情应急防治”悬赏项目(2020254008);深圳市龙岗区科技发展资金“新型冠状病毒感染应急防治”科技专项(LGKCXGZX2020012)。

摘　　要：目的建立新型冠状病毒(2019-nCoV)、甲型流感病毒(Flu A)和乙型流感病毒(Flu B)胶体金抗原联合快速检测试剂的制备工艺,并系统评价试剂的性能指标。方法采用柠檬酸三钠两步还原法制备胶体金溶液,用抗2019-nCoV核衣壳蛋白(NP)单抗、抗Flu A NP单抗、抗Flu B NP单抗及多肽作为标记抗体,硝酸纤维素膜上分别包被抗2019-nCoV NP单抗和抗多肽单抗制备新型冠状病毒抗原试纸条,另一硝酸纤维素膜上分别包被抗Flu A NP单抗、抗Flu B NP单抗和抗多肽单抗制备甲/乙型流感病毒抗原试纸条。对试剂盒最低检出限、重复性、交叉反应及临床诊断符合率进行性能评价。结果检测国家参考品,试剂盒最低检出限和重复性均符合要求;分析特异性研究显示,试剂盒与16种呼吸道相关病原体毒株均无交叉反应性。临床研究显示,检测新型冠状病毒总符合率为99.11%(111/112),甲型流感病毒总符合率为98.78%(81/82),乙型流感病毒诊断总符合率为97.56%(80/82)。结论新型冠状病毒和甲/乙型流感病毒抗原联合快速检测试剂检测灵敏度和特异性高,出结果快,操作便携,可用于新型冠状病毒和甲/乙型流感病毒的快速筛查及鉴别诊断。Objective To establish a immunoassay for the combo rapid detection of antigen against novel coronavirus(2019-nCoV),influenza A(Flu A)and influenza B(Flu B),and to evaluate its clinical performance.Methods The colloidal gold was synthesized by two-step reduction of trisodium citrate.The monoclonal antibodies against 2019-nCoV nucleocapsid protein(NP),Flu A NP,Flu B NP and peptides were labeled with colloidal gold nanoparticles.The monoclonal antibodies of anti-2019-nCoV NP and anti-peptides were immobilized on the nitrocellulose membrane to prepare 2019-nCoV test strip.The other nitrocellulose membrane was immobilized with the monoclonal antibodies of anti-Flu A NP,anti-Flu B NP and anti-peptides to prepare influenza A/B test strip.The performance metrics of the lowest detect limit,repeatability,cross-reactivity,total coincidence rate of clinical diagnosis were evaluated.Results The lowest detect limit and repeatability of the kit for testing national reference materials met the requirements.The analysis specificity study showed that the kit had no cross reactivity with 16 strains of respiratory related pathogens.Clinical research showed that the diagnostic total coincidence rate of 2019-nCoV,influenza A and influenza B were 99.11%(111/112),98.78%(81/82)and 97.56%(80/82),respectively.Conclusion The combined rapid detection reagent of 2019-nCoV and influenza A/B has high sensitivity and specificity,portable operation,fast speed,which can be used for rapid screening and differential diagnosis of 2019-nCoV and influenza A/B virus.

关 键 词：新型冠状病毒 流感病毒 抗原 胶体金 免疫层析 性能评价 

分 类 号：R563.1[医药卫生—呼吸系统]