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首页> 外文期刊>Biosensors & Bioelectronics: The International Journal for the Professional Involved with Research, Technology and Applications of Biosensers and Related Devices >Multiple self-cleaning paper-based electrochemical ratiometric biosensor based on the inner reference probe and exonuclease III-assisted signal amplification strategy
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Multiple self-cleaning paper-based electrochemical ratiometric biosensor based on the inner reference probe and exonuclease III-assisted signal amplification strategy

机译：基于内参考探针和外切核酸酶III辅助信号放大策略的多种自清洁纸基电化学比例生物传感器

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Rapid and accurate detection of nucleic acids plays a major role in biological research and clinical diagnostics. Here, a 3D multiple self-cleaning electrochemical ratiometric microfluidic paper-based analytical device (SER-mu PAD) has been constructed for manganese super oxide dismutase (MnSOD) gene detection on the basis of the inner reference probe and exonuclease III (Exo LLD-assisted analytes recycling amplification method. To simplify manual operations, a multipath self-cleaning tab that could manipulate fluid transport was introduced into the paper-based device, realizing time-programmable self-cleaning of the electrode. For achieving sensitive detection of MnSOD gene, the methylene blue (MB)-modified capture probe (CP) as the inner reference element was first self-assembled on triangular Au nanosheets modified paper working electrode to provide a built-in correction and improve the detection accuracy. When MnSOD gene existed, it hybridized with the hairpin-structured signal probe, triggering the cyclic amplification with the assistance of Exo III selective digestion to engender numerous residual DNA labeled with ferrocene (Fc) that could be captured on electrode surface by CPs. Hence, the Fc tags were close to the electrode surface, resulting in the oxidation peak current of Fc (I-FC) increase, while that of MB (I-MB) was constant on account of the unchanged distance between the MB tags and the electrode. The value of I-FC/I-MB was linear with MnSOD gene concentration from 10 nM to 1200 nM, and the detection limit was 3.91 nM. This strategy provides an accurate, robust, and sensitive method for nucleic acids detection and shows great potential in the construction of portable devices.
机译：快速和准确地检测核酸在生物学研究和临床诊断中起着重要作用。这里，在内参考探针和外切核酸酶III（EXO LLD-辅助分析物回收扩增方法。为了简化手动操作，将流体传输的多径自清洁标签引入基于纸的装置，实现了电极的时间可编程自清洁。为了实现MNSOD基因的敏感性检测，作为内参考元件的亚甲基蓝色（MB）制片捕获探针（CP）首先在三角形Au纳米晶片上自组装，改性纸张工作电极，以提供内置校正并提高检测精度。当MNSOD基因存在时，它用发夹结构信号探针杂交，在EXO III选择性消化的帮助下触发循环扩增，以获得许多RE用二茂铁（Fc）标记的偶联DNA，可通过CPS在电极表面上捕获。因此，Fc标签靠近电极表面，导致Fc（I-Fc）的氧化峰值电流增加，而MB（I-MB）的氧化峰值是恒定的，因为MB标签与...之间的不变距离是恒定的电极。 I-Fc / I-Mb的值与MnSod基因浓度为10nm至1200nm的线性，检测限为3.91nm。该策略为核酸检测提供了准确，稳健和敏感的方法，并且在便携式设备的构造方面具有巨大潜力。

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