Electrochemical genosensor for the rapid detection of GMO using loop-mediated isothermal amplification
Electrochemical genosensor for the rapid detection of GMO using loop-mediated isothermal amplification
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DOI：
10.1039/b812569d
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发表时间：
2009-01-01
期刊：
影响因子：
4.2
通讯作者：
Tamiya, Eiichi
中科院分区：
文献类型：
--
作者：
Ahmed, Minhaz Uddin;Saito, Masato;Tamiya, Eiichi
在这项研究中，我们首次报道了一种高效、准确和廉价的快速检测系统，该系统结合了恒温扩增和电化学系统对未纯化扩增产物的后续分析。在我们的实验中，环介导的等温扩增(LAMP)是在恒温(65℃)下进行的，其效率高于PCR。扩增产物与氧化还原活性分子Hoechst 33258[H33258，2‘-(4-hydroxyphenyl)-5-(4-methyl-1-piperazinyl)-2，5’-bi(1H-benzimidazole)]]结合，用与一次性电化学打印芯片集成的DNA棒(DS)在线性扫描伏安法(LSV)下进行分析。H33258分子的DNA小槽结合导致H33258氧化的峰电流强度显著下降。利用H33258诱导的DNA结合现象以及阳极电流峰的变化，对玉米品种(StarLink(TM))进行了检测。由于不需要费力地固定探针，并且在单个设备上进行扩增和检测，我们的生物传感器消除了潜在的交叉污染。我们相信，这种传感器将对环境保护产生前所未有的影响。
In this study, we are reporting for the first time an efficient, accurate and inexpensive rapid detection system which employs the integration of isothermal amplification and subsequent analysis of unpurified amplicons by an electrochemical system. In our experiments, loop-mediated isothermal amplification (LAMP) with its higher efficiency than PCR was performed at a constant temperature (65 degrees C). Amplification products were combined with a redox active molecule Hoechst 33258 [H33258, 2'-(4-hydroxyphenyl)-5-(4-methyl-1-piperazinyl)-2,5'-bi(1H-benzimidazole)] and analyzed by a DNA stick (DS) which is integrated with a disposable electrochemical printed (DEP) chip using linear sweep voltammetry (LSV). The DNA minor groove binding of the H33258 molecule causes a significant drop in the peak current intensity of the H33258 oxidation. The phenomenon of DNA binding induced by H33258, in addition to changes in the anodic current peak, was used to detect maize CBH 351 variety (StarLink (TM)). Since laborious probe immobilization was not required, and amplification and detection were performed on a single device, our biosensor eliminates potential cross-contamination. We believe that this type of sensor will have an unprecedented impact for environmental protection.