利用F_0F_1-ATPase分子马达生物传感器检测食品中的副溶血性弧菌  

Detection of Vibrio parahaemolyticus in Foods Using F_0F_1-ATPase Molecular Motor Biosensor

作　　者：张捷[1] 顾德周[1] 张惠媛[1] 刘岩[1] 汪琦[1] 张昕[1] 王佩荣[2] 陶雨风[3] 范斐[1] 陈广全[1] 乐加昌[2] 

机构地区：[1]北京出入境检验检疫局,北京100026 [2]中国科学院生物物理研究所,北京100101 [3]中国合格评定国家认可委员会,北京100062

出　　处：《食品科学》2012年第24期226-229,共4页Food Science

基　　金：国家质检总局科技计划项目(2011IK191)

摘　　要：利用载色体上的F0F1-ATPase分子马达生物传感器对副溶血性弧菌检测快速检测。在ATP合酶的ε亚基上连接ε亚基抗体-生物素-链霉亲和素-生物素-toxR探针,将待测副溶血性弧菌标准菌株和阴性对照分别与生物传感器结合后,比较其催化ATP合成30min后的ATP产生量,可以对副溶血性弧菌的DNA进行检测。ATP合成的多寡可以通过环境中H+的量进行测量,而H+的多寡通过F-DHPE所体现的荧光强度大小来获得。结果表明,chro toxR的质量浓度在0.156mg/mL,副溶血性弧菌DNA质量浓度在40ng/mL时为最适检出条件。通过与实际检测样品的传统检测方法及聚合酶链式反应检测方法对照,具有良好的符合性。An F0F1-ATPase molecular motor biosensor was used to rapidly detect Fibrio parahaemolyticus in foods. For the construction of a biosensor, a specific toxR probe was connected to the ε subunit of F0F1-ATPase avidin-biotin system using an avidin-biotin system. The detection of Fibrio parahaemolyticus was based on differences in ATP production 30 rain after separate conjugation of Vibrio parahaemolyticus and negative control to the constructed biosensor. ATP production was determined by measuring the H＋ amount in the environment, which was dependent on the fluorescence intensity of F-DHPE. The optimum concentrations of chro toxR and Vbrio parahaemolyticus DNA for detecting Vbrio parahaemolyticu were found to be 40 ng/mL and 40 ng/mL, respectively. In practical applications, the results obtained by this method were in good agreement with those obtained by traditional detection methods and PCR.

关 键 词：F0F1-ATPase分子马达 生物传感器 副溶血性弧菌 快速检测 

分 类 号：TS207.4[轻工技术与工程—食品科学]