Target-induced reconfiguration of DNA probes for recycling amplification and signal-on electrochemical detection of hereditary tyrosinemia type I gene.
Target-induced reconfiguration of DNA probes for recycling amplification and signal-on electrochemical detection of hereditary tyrosinemia type I gene.
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DOI：
10.1039/c5an01006c
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发表时间：
2015-08
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影响因子：
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通讯作者：
Baoting Dou;Cuiyun Yang;Y. Chai;R. Yuan;Yun Xiang
中科院分区：
文献类型：
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作者：
Baoting Dou;Cuiyun Yang;Y. Chai;R. Yuan;Yun Xiang
通过偶联靶DNA诱导的重组的dsDNA探针与酶辅助的靶循环扩增，我们描述了一个信号的电化学传感方法的发展，用于敏感检测遗传性酪氨酸血症I型基因。dsDNA探针在传感电极上自组装，并且靶DNA的添加将dsDNA探针重新配置并转换为外切核酸酶III的活性底物，这催化地破坏探针并导致靶DNA的循环再利用。靶DNA再循环和通过核酸外切酶III从dsDNA探针中去除一个ssDNA导致在传感器表面上形成许多发夹结构，这使电活性亚甲蓝标记接近电极并产生显著放大的电流响应，用于灵敏地检测低至0.24 pM的靶基因。该方法对单碱基错配也有选择性，可用于人血清样品中目的基因的检测。通过对目标基因的检测，我们期望所开发的方法能够成为一种通用的灵敏传感平台，用于检测不同的核酸序列。
By coupling target DNA-induced reconfiguration of the dsDNA probes with enzyme-assisted target recycling amplification, we describe the development of a signal-on electrochemical sensing approach for sensitive detection of hereditary tyrosinemia type I gene. The dsDNA probes are self-assembled on the sensing electrode, and the addition of the target DNA reconfigures and switches the dsDNA probes into active substrates for exonuclease III, which catalytically digests the probes and leads to cyclic reuse of the target DNA. The target DNA recycling and the removal of one of the ssDNA from the dsDNA probes by exonuclease III result in the formation of many hairpin structures on the sensor surface, which brings the electroactive methylene blue labels into proximity with the electrode and produces a significantly amplified current response for sensitive detection of the target gene down to 0.24 pM. This method is also selective to discriminate single-base mismatch and can be employed to detect the target gene in human serum samples. With the demonstration for the detection of the target gene, we expect the developed method to be a universal sensitive sensing platform for the detection of different nucleic acid sequences.