5种呼吸道病毒Array-ELISA检测方法的建立  

Establishment of Array-ELISA to Detect Five Respiratory Viruses

作　　者：李裕昌[1] 康晓平[1] 张雨[1] 吴晓燕[1] 张雪松[1] 冉鑫[1] 霍耐凡 贾佳[1] 杨银辉[1] 

机构地区：[1]军事医学科学院微生物流行病研究所病原微生物生物安全国家重点实验室,北京100071

出　　处：《生物技术通讯》2015年第3期403-407,共5页Letters in Biotechnology

摘　　要：目的:建立能够同时检测常见呼吸道病毒的Array-ELISA方法。方法:未经标记的单克隆抗体作为捕获抗体在96孔板的微孔中点制成阵列,生物素标记的单克隆抗体作为检测抗体,与待检病毒反应后,利用电化学发光法放大反应信号,通过CCD传感器读取反应结果。结果:经过优化后,使用的抗体组合能够检测甲型流感病毒、乙型流感病毒、呼吸道合胞病毒、腺病毒和副流感病毒3型,其最低检测限分别为3.1×102、1.3×103、5×103、2.5×103和1.5×104TCID50/m L,各病毒之间未见交叉反应。结论:建立的Array-ELISA方法可用于检测以上5种病毒,具有较高的灵敏性和较强的特异性。Objective： To establish a method of detecting common respiratory viruses simultaneously by means ofArray- ELISA.Methods： Unlabeled monoclonal antibody（m Ab） used as capture antibody were printed on eachwell of 96-well plate,biotin labeled m Ab were utilized as detection antibody,after reacting with viral antigens,the reaction signals were amplified through electrochemiluminescence and recorded by CCD sensor.Results： Anti-body pairs utilized can detect IAV,IBV,RSV,ADV and PIV-3,and the limits of detection were 3.1×10^2,1.3×10^3,5×10^3,2.5×10^3and 1.5×10^4TCID50/m L respectively,no cross reaction was observed.Conclusion： The estab-lished Array-ELISA can detect five viruses simultaneously,with relatively high sensitivity and specificity.

关 键 词：Array-ELISA 甲型流感病毒 乙型流感病毒 呼吸道合胞病毒 腺病毒 副流感病毒3型 检测 

分 类 号：Q78[生物学—分子生物学]