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首页> 外文期刊>ACS applied materials & interfaces >In Situ DNA-Templated Synthesis of Silver Nanoclusters.for Ultrasensitive and Label-Free Electrochemical Detection of MicroRNA
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In Situ DNA-Templated Synthesis of Silver Nanoclusters.for Ultrasensitive and Label-Free Electrochemical Detection of MicroRNA

机译：银纳米簇的原位DNA模板合成用于MicroRNA的超灵敏和无标记电化学检测

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摘要

On the basis of the use of silver nanoclusters (AgNCs) in situ synthesized by cytosine (C)-rich loop DNA templates as signal amplification labels, the development of a label-free and highly sensitive method for electrochemical detection of microRNA (miRNA-199a) is described. The target miRNA-199a hybridizes with the partial dsDNA probes to initiate the target-assisted polymerization nicking reaction (TAPNR) amplification to produce massive intermediate sequences, which can be captured on the sensing electrode by the self-assembled DNA secondary probes. These surface-captured intermediate sequences further trigger the hybridization chain reaction (HCR) amplification to form dsDNA polymers with numerous C-rich loop DNA templates on the electrode surface. DNA-templated synthesis of AgNCs can be realized by subsequent incubation of the dsDNA polymer-modified electrode with AgNO3 and sodium borohydride. With this integrated TAPNR and HCR dual amplification strategy, the amount of in situ synthesized AgNCs is dramatically enhanced, leading to substantially amplified current response for highly sensitive detection of miRNA-199a down to 0.64 fM. In addition, the developed method also shows high selectivity toward the target miRNA-199a. Featured with high sensitivity and label-free capability, the proposed sensing scheme can thus offer new opportunities for achieving sensitive, selective, and simple detection of different types of microRNA targets.
机译：在使用富含胞嘧啶（C）的环DNA模板原位合成的银纳米团（AgNC）作为信号放大标记的基础上，开发了一种无标记且高度灵敏的电化学检测microRNA（miRNA-199a）的方法）进行了说明。靶标miRNA-199a与部分dsDNA探针杂交，以启动靶标辅助聚合切口反应（TAPNR）扩增，以产生大量的中间序列，该序列可由自组装的DNA二级探针捕获在传感电极上。这些表面捕获的中间序列进一步触发了杂交链反应（HCR）扩增，以形成在电极表面具有大量富C环DNA模板的dsDNA聚合物。通过随后将dsDNA聚合物修饰的电极与AgNO3和硼氢化钠一起孵育，可以实现AgNCs的DNA模板合成。通过这种集成的TAPNR和HCR双重扩增策略，原位合成AgNCs的数量得到显着提高，从而导致对miRNA-199a的高灵敏度检测的电流反应得到了大幅放大，降低至0.64 fM。另外，开发的方法还显示出对靶miRNA-199a的高选择性。所提出的传感方案具有高灵敏度和无标记的功能，因此可以为实现不同类型的microRNA靶标的灵敏，选择性和简单检测提供新的机会。

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