F_0F_1-ATPase分子马达技术对单核细胞增生李斯特菌检测的研究  

The Study of a Detecting Technology for Listeria monocytogenes based on F_0F_1-ATPase Molecular Motor Biosensor

作　　者：张捷[1] 张昕[1] 范爱红 张惠媛[1] 汪琦[1] 顾德周[1] 王佩荣[3] 韩晶[1] 陆琳[1] 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]国家认监委认证认可技术研究所,北京100020 [3]中国科学院生物物理研究所,北京100101

出　　处：《生物技术进展》2012年第6期436-440,共5页Current Biotechnology

基　　金：国家质检总局科技计划项目(2008IK161)资助

摘　　要：利用载色体chromatophore上的F0F1-ATPase分子马达生物传感器,建立了应用在食品检测中快速检测方法。首先从嗜热菌中提取载色体后,标记荧光探针F-DHPE,合成生物素化单增李氏菌prfA探针,在已标记F-DHPE的载色体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-prfA探针,将待测单核细胞增生李斯特菌标准菌株和阴性对照分别与此生物传感器结合,通过环境H+量测定ATP产生量,进而对单核细胞增生李斯特菌DNA进行检测。结果表明,chroprfA(连接在载色体chro上的prfA探针)的浓度在0.052mg/mL,单核细胞增生李斯特菌DNA浓度在40ng/mL为最适检测条件。通过传统检测方法及PCR检测方法对照,本方法具有良好的检测符合性。A rapid detecting technology was constructed by using chromatophore on the F0F1-ATPase molecular motor biosensor for Listeria monocytogenes in the food. Specific prfA probe were connected with F0F1-ATPase’s ε subunit by using avidin-biotin system, the test samples and negative sample combined with biosensors, respectively, to compare their ATP synthesis by measuring the amount of H＋, Listeria monocytogenes DNA in the samples could be tested. The results showed that optimum conditions for detection was using chro prfA concentration of 0.052 mg/mL and Listeria monocytogenes DNA concentration of 40 ng/mL. By contrasting to conventional detection methods and PCR detection method to test the actual samples, this method has good conformity.

关 键 词：F0F1-ATPase分子马达 单核细胞增生李斯特菌 prfA探针 快速检测 

分 类 号：R155.5[医药卫生—营养与食品卫生学]