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Development of Bacteria sensor using Dielectrophoresis and Electropermeabilization

利用介电泳和电透化技术开发细菌传感器

基本信息

批准号：
16360207
负责人：
SUEHIRO Junya
金额：
$ 9.6万
依托单位：
KYUSHU UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2004
资助国家：
日本
起止时间：
2004 至 2005
项目状态：
已结题

项目摘要

Electropermeabilization (EP) was successfully incorporated with DEPIM in order to achieve higher sensitivity for bacteria detection. Contamination of metallic ions released from electrodes, which masks subtle conductance increase caused by the released intracellular ions, was successfully suppressed by improving the pulse waveform for EP. E. coli was quantitatively detected in 3 h at 10^2 CFU/ml, which was improved by two orders of magnitude relative to the conventional DEPIM without EP. The proposed technique is promising as a fast and sensitive bacteria inspection method and may be easily incorporated into Bio-MEMS or μ-TAS devices.In order to add selectivity to DEPIM or EPA-DEPIM inspection of bacteria, new inspection protocols were demonstrated using an electrode chip on which a target bacteria-specific antibody was immobilized in advance. In the first method, all bacteria were first trapped onto the electrode chip by strong positive DEP. In order to selectively retain the target bacteria, which could bind with the immobilized antigen, the hydrodynamic drag force was exerted by washing liquid flow. Although the number of retained target bacteria increased by using the antibody-modified chip, undesired non-specific binding to the unmodified chip also took place. The second method utilized lower DEP force so that trapping and washing processes could be simultaneously accomplished. A smaller number of retained bacteria under decreased DEP force was surely detected by EPA-DEPIM, which was more sensitive than the conventional DEPIM. It was found that this protocol could also suppress the non-specific bacteria binding and greatly improve the selectivity for the target bacteria.

为了获得更高的细菌检测灵敏度，成功地将电渗透（EP）与DEPIM结合。电极释放的金属离子污染掩盖了细胞内离子释放引起的细微电导增加，通过改善脉冲电位波形成功地抑制了污染。在10^2 CFU/ml下，3 h即可定量检测出大肠杆菌，相对于不加EP的常规DEPIM提高了两个数量级。该技术是一种快速、灵敏的细菌检测方法，可以很容易地集成到Bio-MEMS或μ-TAS器件中。为了增加细菌的DEPIM或EPA-DEPIM检测的选择性，使用预先固定目标细菌特异性抗体的电极芯片演示了新的检测方案。在第一种方法中，首先通过强正DEP将所有细菌捕获到电极芯片上，通过洗涤液流动施加水动力阻力，以选择性保留可与固定化抗原结合的目标细菌。虽然通过使用抗体修饰的芯片，保留的目标细菌数量增加了，但与未修饰芯片的非特异性结合也发生了。第二种方法利用较低的DEP力，使捕集和洗涤过程可以同时完成。EPA-DEPIM比常规DEPIM更敏感，在DEP力降低的情况下，残留细菌数量明显减少。发现该方案还可以抑制非特异性细菌的结合，大大提高了对目标细菌的选择性。

项目成果

期刊论文数量（20）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Controlled fabrication of carbon nanotube NO2 gas sensor using dielectrophoretic impedance measurement

DOI：
10.1016/j.snb.2004.09.048
发表时间：
2005-07
期刊：
Sensors and Actuators B-chemical
影响因子：
8.4
作者：
J. Suehiro;Guangbin Zhou;Hiroshi Imakiire;W. Ding;M. Hara
通讯作者：
J. Suehiro;Guangbin Zhou;Hiroshi Imakiire;W. Ding;M. Hara

Electropermiabilization-assested dielectrophoretic impedance measurement method for high sensitive bacteria detection

用于高灵敏度细菌检测的电透化介电泳阻抗测量方法

DOI：
发表时间：
2004
期刊：
Proc. of Int.Conf. on Electrical Bioimpedance and Electrical Impedance Tomography
影响因子：
0
作者：
J.Suehiro;M.Shutou;T.Hatano;M.Hara
通讯作者：
M.Hara

Electropermeabilization-assisted dielectrophoretic impedance measurement method for high sensitive bacteria detection

高灵敏细菌检测的电透辅助介电泳阻抗测量方法

DOI：
发表时间：
2004
期刊：
Proc. of Int. Conf. on Electrical Bioimpedance and Electrical Impedance Tomography
影响因子：
0
作者：
J.Suehiro;M.Shutou;T.Hatano;M.Hara
通讯作者：
M.Hara

Improvement of electric pulse shape for electropermeabilization-assisted dielectrophoretic impedance measurement for high sensitive

高灵敏度电透辅助介电泳阻抗测量电脉冲形状的改进