利用分子马达技术检测霍乱弧菌  

Detecting of Vbrio cholerae Using Molecular Motor Biosensor

作　　者：张捷[1] 向丽萍 汪琦[1] 张惠媛[1] 张昕[1] 王宇 顾德周[1] 王佩荣[4] 温铮[1] 陈广全[1] 乐加昌[4] 

机构地区：[1]北京出入境检验检疫局,北京100026 [2]遵义市产品质量检验检测院,贵州遵义563000 [3]国家认监委认证认可技术研究所,北京100020 [4]中国科学院生物物理研究所,北京100101

出　　处：《食品与发酵工业》2012年第6期161-164,共4页Food and Fermentation Industries

基　　金：国家质检总局科技计划项目(2010IK156)

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的快速检测方法。首先在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-ompW探针,将待测霍乱弧菌和阴性对照分别与此生物传感器结合,比较其催化ATP合成30 min后的ATP产生量,进而对霍乱弧菌的DNA进行检测。ATP合成的量可以通过环境H+的量进行测定,H+的量通过F-DHPE所体现的荧光强度大小标定。结果表明,Chro ompW的浓度在0.078 mg/mL,单核增生李斯特菌DNA浓度在40 ng/mL为最适检测条件。通过与实际检测样品的传统检测方法及PCR检测方法对照,具有良好的检测符合性。The core technology was that the F0F1-ATPase molecular motor biosensor on chromatophore was used to rapidly detect Vbrio cholerae.Specific ompW probe were connected with ε subunit of F0F1-ATPase by avidin-biotin system,and then biosensors were constructed,and the test samples and negative sample were respectively combined with biosensors.After the ATP synthesis was catalyzed by them for 30 min,the content of ATP and Vbrio cholerae DNA in the samples were measured.The amount of ATP synthesis could be measured via the measurement of the amount of H＋,while the amount of H＋ was normalized by the fluorescence intensity of F-DHPE.According to the results of our experiments,optimum conditions for detection was under the presence of 0.078 mg/mL chro ompW and 40 ng/mL Vbrio cholerae DNA.After detection of the actual samples,we found that the method was in good agreement with the traditional detection methods and PCR detection method.

关 键 词：F0F1-ATPase分子马达 霍乱弧菌 ompW探针 快速检测 

分 类 号：R155.5[医药卫生—营养与食品卫生学]