F_0F_1-ATPase生物传感器检测食品中食源性诺如病毒  

The Establishment of Molecular Motor Biosensor Technologies Rapid Detection Method for Norovirus in Food

作　　者：张捷[1] 许美玲 王璇[3] 王煜[4] 刘岩[1] 王小晋[5] 张丽 张慧媛[1] 顾德周[1] 王佩荣[3] 陈广全[1] 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：《食品研究与开发》2014年第24期116-120,共5页Food Research and Development

基　　金：国家质检总局科技计划项目(2010IK156)

摘　　要：应用分子马达生物传感技术,建立食品中诺如病毒特异性快速检测方法。以嗜热菌中提取的载色体(chromatophore)为材料,根据诺如病毒ORF1和ORF2连接处的高度保守区设计探针,利用生物素-亲和素系统将诺如病毒探针连接在F0F1-ATPase的ε亚基上构建生物传感器。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,对样品中的RNA进行检测。建立并优化了F0F1-ATPase生物传感技术检测食源性诺如病毒的方法。所建立的方法具有良好的特异性,12个含诺如病毒样品均能检出,3个不含诺如病毒样品均未检出,无假阴性或假阳性情况出现。该方法可在1 h内完成检测,检测灵敏度在RNA水平上达到0.005 ng/m L。所建立的分子马达生物传感技术可以较好的检测食品中的诺如病毒,并为其他病毒的检测提供参考。This study is to establish rapid detection method for Norovirus in food using molecular motor biosensor technologies. Chromatophores are inverted plasma membrane vesicles containing F0F1-ATPase, and the chromatophores were used as a model system to detect Norovirus. A specific probe was encompassed on the conservative region between ORF1 and ORF2. A biosensors were constructed by connecting specific molecular probes with F0F1-ATPase＇sεsubunit throughavidin-biotin system. Extracted virus RNA was conjugated with the biosensor separately, at the same time ATP were synthesized. By comparing their fluorescence intensity, we can detect Norovirus RNA in samples. Through optimizing the reaction condition, a simple and rapid method for detection of Noroviris was developed by F0F1-ATPase biosensor. The results of the study indicated that the method had perfect specificity, and analysis of Hepatitis A visus and Rotavirus demonstrated that this system was specific. The test could be done within 1 h with the sensitivity low to 0.005 ng/mL with genomic RNA. Fifteen samples artificially contaminated and fifty samples were detected, and no false positive or false negative results. Result indicated that this reliable, sensitive, quick and easy-handling method could be used for the detection of food-borne Norovirus virus, and could also be a valuable reference to other food-borne viruses.

关 键 词：食源性病毒 诺如病毒 分子马达生物传感器 快速检测 

分 类 号：TP212.3[自动化与计算机技术—检测技术与自动化装置] TS207.4[自动化与计算机技术—控制科学与工程]