一种快速检测甲型流感病毒方法的建立  

Establishment of a rapid method for detecting influenza A virus

作　　者：朱元娅 刁维均 胡宏章[3] ZHU Yuan-ya;DIAO Wei-jun;HU Hong-zhang(Chengdu Shang Jin Nan Fu Hospital,Chengdu,Sichuan 6111431 China;Maccura Biotechnology Co.,Ltd.,Chengdu,Sichuan 611731,China;Department of Laboratory Medicine,West China Hospital of Sichuan University,Chengdut Sichuan 610041,China)

机构地区：[1]成都上锦南府医院,四川成都611743 [2]迈克生物股份有限公司,四川成都611731 [3]四川大学华西医院实验医学科,四川成都610041

出　　处：《实用预防医学》2020年第3期379-382,共4页Practical Preventive Medicine

摘　　要：目的建立一种用于甲型流感病毒抗原检测的基于胶体金免疫层析法的快速、简便试剂盒。方法以柠檬酸三钠还原法制备胶体金颗粒,标记一种抗甲型流感病毒单克隆抗体。硝酸纤维素膜上包被另一种抗甲型流感病毒单克隆抗体,制成免疫层析试纸。待测样品中的甲型流感病毒首先与胶体金标记抗体结合,然后移动至硝酸纤维素膜上与固定的单克隆抗体发生反应,形成肉眼可见的红色带。结果本试纸与不同浓度甲型流感病毒培养液均能发生特异性反应,与乙型流感病毒、呼吸道合胞病毒、副流感病毒等16种病原体无交叉反应。用甲型流感病毒培养液不同浓度标本与美国同类产品比较,灵敏度相同。稳定性检测显示42℃和37℃热破坏放置结果均优于美国同类产品。临床比对结果评价kappa值为0.976,一致性非常高。结论试纸条灵敏度和特异性能够达到临床使用的要求,并具有简便快速、无需特殊仪器设备等优点,对甲型流感的诊断和流行病学调查具有十分重要的应用价值。Objective To establish a rapid and simple kit based on the colloidal gold immunochromatography assay(GICA)for the detection of human influenza A virus antigen.Methods Colloidal gold particles were prepared by the method of trisodium citrate reduction,and labeled with a monoclonal antibody against influenza A virus.The cellulose nitrate membrane was coated with another monoclonal antibody against influenza A virus and made into an immunochromatographic test strip.Influenza A viruses in the tested samples were firstly bound to colloidal gold labeled antibodies,and then moved to nitrocellulose membrane to react with fixed monoclonal antibodies and form a visible red band.Results The test strips could react specifically with different concentration of influenza A virus culture medium,and did not cross-react with 16 pathogens,such as influenza B virus,respiratory syncytial virus and parainfluenza virus.The sensitivities of the different concentration of the influenza A virus culture solution were the same as those of the similar products made in the USA.Stability detection displayed that 42℃and 37℃thermal destruction placement results were superior to those of the similar products made in the USA.Comparison and evaluation of clinical results showed that the Kappa value was 0.976,and the consistency was very high.Conclusions The sensitivity and specificity of the test strip can meet the requirement of clinical use;moreover,it has the advantages of simplicity and rapidness with no need for special apparatus and equipment.The test strip has important application value for the diagnosis and epidemiological investigation of influenza A.

关 键 词：胶体金免疫层析法 甲型流感病毒 特异性 灵敏度 

分 类 号：R511.7[医药卫生—内科学]