Detection of SEB gene by bilayer lipid membranes nucleic acid biosensor supported by modified patch-clamp pipette electrode.
Detection of SEB gene by bilayer lipid membranes nucleic acid biosensor supported by modified patch-clamp pipette electrode.
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DOI：
10.1016/j.bios.2006.09.037
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发表时间：
2007-04
影响因子：
12.6
通讯作者：
Nan Liu;Zhixian Gao;Huanying Zhou;Min-Zhi Yue
中科院分区：
文献类型：
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作者：
Nan Liu;Zhixian Gao;Huanying Zhou;Min-Zhi Yue
报道了一种新型的膜片钳电极支撑的双层脂膜核酸传感器，用于检测葡萄球菌肠毒素B(SEB)基因。BLMS在15min内形成，至少可操作24小时。将疏水十二烷尾(C12)修饰的18bp单链DNA(SsDNA)探针固定在BLMS上。电化学电流与固定在BLMS上的不同浓度的单链DNA探针呈线性相关。选择浓度为273.65 ng/mL的单链DNA探针作为信号传感元件，制备了BLMS核酸生物传感器。研究了该生物传感器检测SEB的电化学性能。结果表明，电流与ln浓度在20~5000 ng/m L范围内呈良好的线性关系，检出限为20 ng/m L。此外，该生物传感器对SEB基因有特异性的响应，在不含SEB基因的电解液中没有明显的电流变化。最后，用原子力显微镜(AFM)观察和评价了BLM、固定在BLM上的单链DNA和杂交后BLM的表面微结构。修饰膜片钳电极支撑的BLMS核酸生物传感器将成为检测产生SEB的金黄色葡萄球菌的一种高灵敏度、快速、选择性的分析工具。
This work reports a kind of novel bilayer lipid membranes (BLMs) nucleic acid biosensor supported by modified patch-clamp pipette electrode was developed to detect staphylococcus enterotoxins B (SEB) gene. BLMs were formed within 15min and able to be operated at least 24h. Hydrophobic dodecane tail (C12) modified 18bp single-stranded DNA (ssDNA) probe was immobilized on BLMs. The electrochemical currents versus the different concentration of ssDNA probe immobilized on BLMs indicated linearly correlation. The BLMs nucleic acid biosensor was fabricated by selecting the ssDNA probe as the signal sensing element with the concentration of 273.65ng/mL. The electrochemical performance of the biosensor for the detection of SEB was investigated. The result showed that linear relationship was found between the current and ln(concentration) from 20 to 5000ng/mL and the detection limit was 20ng/mL. In addition, the biosensor was specific response to SEB gene and showed no significant current alteration in electrolyte which containing no SEB gene. Finally, Atom Force Microscope (AFM) images could be observed and used to evaluate the superficial microstructure of BLMs, ssDNA immobilized on BLMs and BLMs after hybridization. The BLMs nucleic acid biosensor supported by modified patch-clamp pipette electrode will become a highly sensitive, rapid, selective analytical tool for detection of Staphylococcus aureus, which produce SEB.