F0F1-ATPase分子马达技术对单核细胞增生李斯特菌检测的研究

doi:10.3969/j.issn.2095-2341.2012.06.09

生物技术进展 ›› 2012, Vol. 2 ›› Issue (6): 436-440.DOI: 10.3969/j.issn.2095-2341.2012.06.09

F₀F₁-ATPase分子马达技术对单核细胞增生李斯特菌检测的研究

张捷1,张昕1,范爱红2,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

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

收稿日期:2012-10-09 出版日期:2012-11-25 发布日期:2012-11-05
作者简介:张捷,博士,高级工程师,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(2008IK161)资助。

The Study of a Detecting Technology for Listeria monocytogenes based on F₀F₁-ATPase Molecular Motor Biosensor

ZHANG Jie, ZHANG Xin, FAN Ai-hong, ZHANG Hui-yuan, WANG Qi, GU De-zhou, WANG Pei-rong, HAN Jing, LU Lin, CHEN Guang-quan, YUE Jia-chang

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

2.China Certification & Accreditiation Institute, Beijing 100020, China;

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

Received:2012-10-09 Online:2012-11-25 Published:2012-11-05

摘要/Abstract

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,chro prfA(连接在载色体chro上的prfA探针)的浓度在0.052 mg/mL,单核细胞增生李斯特菌DNA浓度在40 ng/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。

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

Abstract: 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.

Key words: F₀F₁-ATPase molecular motor, Listeria monocytogenes, prfA probe, rapid detection

张捷,张昕,范爱红,张惠媛,汪琦,顾德周,王佩荣,韩晶,陆琳,陈广全,乐加昌. F₀F₁-ATPase分子马达技术对单核细胞增生李斯特菌检测的研究[J]. 生物技术进展, 2012, 2(6): 436-440.

ZHANG Jie1, ZHANG Xin1, FAN Ai-hong2, ZHANG Hui-yuan1, WANG Qi1, GU De-zhou1, WANG Pei-rong3, HAN Jing1, LU Lin1, CHEN Guang-quan1, YUE Jia-chang3. The Study of a Detecting Technology for Listeria monocytogenes based on F₀F₁-ATPase Molecular Motor Biosensor[J]. Curr. Biotech., 2012, 2(6): 436-440.

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F₀F₁-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 F₀F₁-ATPase Molecular Motor Biosensor

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