F_0F_1-ATPase分子马达技术对阪崎肠杆菌检测的研究  

A Rapid Detecting Method for Enterobacter sakazakii based on F_0F_1-ATPase Molecular Motor Biosensor

作　　者：张捷[1] 王煜[2] 陆琳[1] 刘艳华[1] 周琦[1] 孙梓晏[1] 顾德周[1] 尚世进[1] 张惠媛[1] 王佩荣[3] 陈广全[1] 乐加昌[3] 

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

出　　处：《生物技术进展》2013年第4期297-300,共4页Current Biotechnology

基　　金：国家质检总局科技计划项目(2012IK146);北京市科委阶梯计划项目资助

摘　　要：本研究建立了应用F0F1-ATPase分子马达生物传感器快速检测阪崎肠杆菌的方法。自嗜热菌中提取载色体后,合成针对阪崎肠杆菌的生物素化ITS探针,在载色体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-ITS探针,将待测阪崎肠杆菌标准菌株和阴性对照分别与此生物传感器结合,比较其催化ATP产生量,进而对阪崎肠杆菌DNA进行检测。结果表明,chro ITS探针浓度0.019mg/mL,阪崎肠杆菌DNA浓度40ng/mL为最适检测条件。通过与传统检测方法及PCR检测方法对照,本方法具有良好的检测符合性。A rapid detecting method for Enterobacter sakazakii based on F0F1 -ATPase molecular motor biosensor was cons tructed. Specific ITS probe were connected with F0 Fl-ATPase＇s 8 subunit by using avidin-biotin system, and then biosensors were combined with the test samples and negative sample, respectively. To compare the catalytic ATP synthesis, Enterobacter sakazakii DNA in the samples were tested. The result showed that the optimum conditions for detection were the concentration of chro ITS of 0. 019 mg/mL and Enterobacter sakazakii DNA concentration of 40 ng/mL. The result was consistent to that of the traditional detection methods and PCR detection method, showing this is in good agreement.

关 键 词：F0F1-ATPase分子马达 阪崎肠杆菌 ITS探针 快速检测 

分 类 号：R155.5[医药卫生—营养与食品卫生学]