一种快速检测甲型/乙型流感病毒抗原方法的建立

Establishment of a rapid method for detection of influenza A/B virus′antigens

本研究探讨建立一种基于量子点免疫层析法联检甲型流感病毒、乙型流感病毒抗原的快速且简便的甲流/乙流联检试剂卡。甲流/乙流联检试剂卡由试纸条和塑料外壳构成,试纸条由吸水纸、缓冲垫、硝酸纤维素膜(NC膜)、样品垫、量子点标记物垫、聚氯乙烯板(PVC板)等组成。硝酸纤维素膜T线(检测线)包被有鼠抗甲型流感病毒单克隆抗体和鼠抗乙型流感病毒单克隆抗体,C线(质控线)包被有内参蛋白A和内参蛋白B,标记物垫上含有鼠抗甲型流感病毒抗体量子点结合物和鼠抗乙型流感病毒抗体量子点结合物。结果显示使用甲流/乙流联检试剂卡,各病毒的检出为1.51×10^(2)~2.71×10^(3) TCID50/ml,证明该试剂的灵敏度足以用于甲流、乙流的精准检测且不受多种变体的影响。本试剂卡与不同分型的甲型流感病毒培养液、不同分型的乙型流感病毒培养液均能发生特异性反应,与腺病毒、新型冠状病毒、肺炎支原体、呼吸道合胞病毒、金黄色葡萄球菌等多种病原体无交叉反应。试剂的灵敏度足以用于甲流、乙流的精准检测且不受多种变体的影响。综上,甲流/乙流联检试剂卡灵敏度和特异性可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,可进行快速辅助诊断,以预防疾病传播。

This study aims to develop a rapid and convenient test card for simultaneous detection of influenza A and influenza B viruses using quantum dot-based immunochromatographic assay.The test card consists of a test strip and a plastic casing.The test strip is composed of absorbent paper,a buffer pad,nitrocellulose membrane(NC membrane),sample pad,quantum dot-labeled antibody pad,and polyvinyl chloride(PVC)board.The NC membrane is coated with mouse monoclonal antibodies against influenza A and influenza B viruses for the T lines(test lines),and reference proteins A and B for the C line(control line).The quantum dot-labeled antibody pad contains mouse monoclonal antibody-quantum dot conjugates against influenza A and influenza B viruses.The results showed that the detection limit of the test card for both viruses ranged from 1.51×10^(2) to 2.71×10^(3) TCID50/ml,indicating its sensitivity for accurate detection of influenza A and influenza B viruses without being affected by various variants.The test card exhibited specific reactions with different subtypes of influenza A and influenza B virus culture fluids and showed no cross-reactivity with adenovirus,novel coronavirus,Mycoplasma pneumoniae,respiratory syncytial virus,Staphylococcus aureus,and other pathogens.Overall,the sensitivity and specificity of the test card for simultaneous detection of influenza A and influenza B viruses meet the requirements for clinical use.It offers the advantages of simplicity,rapidity,and no requirement for special equipment,enabling quick auxiliary diagnosis to prevent disease transmission.