利用F_0F_1-ATPase免疫生物传感器快速检测人肠道病毒71型的研究  

Research on rapid detection of EV71 with F_0F_1-ATPase immune biosensor

作　　者：欧新华[1] 孙边成[1] 王佩荣[2] 陈静芳[1] 乐加昌[2] 

机构地区：[1]长沙市疾病预防控制中心,湖南长沙410001 [2]中国科学院生物物理研究所,北京100101

出　　处：《中国卫生检验杂志》2015年第12期1932-1934,1940,共4页Chinese Journal of Health Laboratory Technology

基　　金：湖南省医药卫生科研计划项目(B2013-126)

摘　　要：目的从嗜热菌中提取载色体Chromatophore,利用F0F1-ATPase构建一种生物传感器,用于人肠道病毒71型(enterovirus 71,EV71)的快速、简便、特异检测。方法制备F0F1-ATPaseε亚基单克隆抗体,通过亲和素-生物素系统的连接作用,以EV71抗体为检测工具,构建一种新的F0F1-ATPaseε亚基单克隆抗体-生物素-链霉亲和素-生物素-EV71抗体的免疫生物传感器,通过比较其检测前后ATP催化合成量的不同,实现对EV71病毒的快速检测。同时,合成用于EV71检测的特异性引物探针,比较2种检测方法的结果。结果 RT-PCR检测灵敏度为10-7mg/ml,检测时间约3 h;生物传感器检测灵敏度为10-7mg/ml,但整个实验过程仅需30 min。结论利用F0F1-ATPase构建的免疫生物传感器可用于EV71感染的快速检测,具有一定的应用前景。Objective From a thermophilic bacteria to extract load color body,chromatophore,and to construct a biosensor with F0F1- ATPase for the rapid,simple and specific detection of human EV71. Methods To prepare F0F1- ATPase epsilon subunit monoclonal antibody,by affinity avidin biotin system connection,using EV71 polyclonal antibody for detecting tool to construct a new F0F1- ATPase epsilon subunit monoclonal antibody biological biotin streptavidin biotin- EV71 antibody immune biosensor,and through comparing the difference between before and after the detection ATP catalytic synthesis to realize the rapid detection of EV71 virus. At the same time,the specificity primer probe for the detection of EV71 was synthesized and the results of the two methods were compared. Results With RT- PCR,the detection sensitivity of EV71 was 10- 7mg / ml and the detection time was about 3 h; with biosensor,the detection sensitivity was also 10- 7mg / ml,but the whole detection process required only about 30 min. Conclusion The constructed biosensor based on F0F1- ATPase could be used in EV71 rapid detection,and this method had promising application prospect.

关 键 词：F0F1-ATPase 生物传感器 人肠道病毒71型 

分 类 号：R373.2[医药卫生—病原生物学]