F0F1-ATPase分子马达技术对阪崎肠杆菌检测的研究

doi:10.3969/j.issn.2095-2341.2013.0 .

生物技术进展 ›› 2013, Vol. 3 ›› Issue (4): 297-300.DOI: 10.3969/j.issn.2095-2341.2013.0 .

• 技术与方法 • 上一篇

F₀F₁-ATPase分子马达技术对阪崎肠杆菌检测的研究

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

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

收稿日期:2013-03-26 出版日期:2013-07-25 发布日期:2013-05-10
作者简介:张捷，高级工程师，博士，研究方向为食品检测和食品安全评价。E-mail：zhangjie@bjciq.gov.cn
基金资助:
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。

A Rapid Detecting Method for Enterobacter sakazakii based on F₀F₁-ATPase Molecular Motor Biosensor

ZHANG Jie, WANG Yu, LU Lin, LIU Yan-hua, ZHOU qi, SUN Zi-yan, GU De-zhou, SHANG Shi-jin, ZHANG Hui-yuan, WANG Pei-rong, CHEN Guang-quan, YUE Jia-chang

1.Beijing Entry-Exit Inspection and Quarantine Bureau, Beijing 100026, China;

2.China National Accreditiation Service for Conformity Accessment, Beijing 100062, China;

3.Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101, China

Received:2013-03-26 Online:2013-07-25 Published:2013-05-10

摘要/Abstract

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0.019 mg/mL，阪崎肠杆菌DNA浓度40 ng/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。

关键词: F₀F₁-ATPase分子马达, 阪崎肠杆菌, ITS探针, 快速检测

Abstract: A rapid detecting method for Enterobacter sakazakii based on F₀F₁-ATPase molecular motor biosensor was cons tructed. Specific ITS probe were connected with F₀F₁-ATPase's ε 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.

Key words: F₀F₁-ATPase , molecular motor, Enterobacter sakazakii; ITS probe, rapid detection

张捷,王煜,陆琳,刘艳华,周琦,孙梓晏,顾德周,尚世进,张惠媛,王佩荣,陈广全,乐加昌. F₀F₁-ATPase分子马达技术对阪崎肠杆菌检测的研究[J]. 生物技术进展, 2013, 3(4): 297-300.

ZHANG Jie1, WANG Yu2, LU Lin1, LIU Yan-hua1, ZHOU qi1, SUN Zi-yan1, GU De-zhou1, SHANG Shi-jin1, ZHANG Hui-yuan1, WANG Pei-rong3, CHEN Guang-quan1, YUE Jia-chang3. A Rapid Detecting Method for Enterobacter sakazakii based on F₀F₁-ATPase Molecular Motor Biosensor[J]. Curr. Biotech., 2013, 3(4): 297-300.

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F₀F₁-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 F₀F₁-ATPase Molecular Motor Biosensor

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