分子马达生物传感器检测食源性轮状病毒  

Detection of food-borne rotavirus by molecular motor biosensor

作　　者：张捷[1] 许美玲 王璇[3] 王煜[4] 王小晋[5] 刘岩[1] 顾德周[1] 陈广全[1] 王佩荣[6] 乐加昌[6] 

机构地区：[1]北京出入境检验检疫局,北京100026 [2]临沂出入境检验检疫局,山东临沂276034 [3]中国中医科学院望京医院检验科,北京100102 [4]中国合格评定国家认可中心,北京100062 [5]淮安出入境检验检疫局,江苏淮安223001 [6]中国科学院生物物理研究所,北京100101

出　　处：《生物工程学报》2013年第5期681-690,共10页Chinese Journal of Biotechnology

基　　金：国家质检总局科技计划项目(No.2010IK156);北京市科委阶梯计划工作项目资助~~

摘　　要：通过分子马达生物传感器技术建立一种特异、便捷、快速的食源性轮状病毒检测方法。以F0F1-ATPase为核心构建分子马达,以轮状病毒保守片段VP7设计各血清型通用探针,通过生物素-亲和素系统将探针与分子马达连接构建F0F1-ATPase分子马达检测装置。提取病毒RNA并将其与生物传感器结合的同时启动ATP合成,比较其荧光强度的差别,可以对样品中的RNA进行检测。此方法的病毒RNA检测灵敏度为0.005 ng/mL,对轮状病毒检测特异,与甲肝病毒、诺如病毒无交叉反应,在1 h内即可完成检测。运用此方法随机检测15份样品,检测结果与RT-PCR一致。结果表明,分子马达生物传感器检测轮状病毒的方法灵敏、特异,可用于食源性轮状病毒的快速检测。To develop a specific, rapid and convenient method based on molecular motor biosensor to detect food-borne rotavirus. A specific probe was encompassed the conservative region of rotavirus＇s VP7 segment, and a molecular motor detect device was constructed by connecting probes to FoF1-ATPase molecular motor through biotin-streptavidin system. This biosensor＇s sensitivity was 0.005 ng/mL for rotavirus RNA. Extracted virus RNA was conjugated with the biosensor separately, at the same time ATP was synthesized. By comparing fluorescence intensity, we can detect rotavirus RNA in samples. This method possessed specificity for rotavirus, without any cross-reaction with Hepatitis A virus and noroviris, and it could be accomplished within 1 h. We detected 15 samples using this method and the results were compared with RT-PCR results. This method is sensitive and specific for rotavirus, and it can be used to detect food-borne rotavirus.

关 键 词：轮状病毒 分子马达生物传感器 检测 

分 类 号：R373[医药卫生—病原生物学]