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Fluorescence turn-on detection of target sequence DNA based on silicon nanodot-mediated quenching

机译:基于硅纳米型介导的淬火的靶序列DNA的荧光导数

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摘要

We have developed a new enzyme-free method for target sequence DNA detection based on the dynamic quenching of fluorescent silicon nanodots (SiNDs) toward Cy5-tagged DNA probe. Fascinatingly, the water-soluble SiNDs can quench the fluorescence of cyanine (Cy5) in Cy5-tagged DNA probe in homogeneous solution, and the fluorescence of Cy5-tagged DNA probe can be restored in the presence of target sequence DNA (the synthetic target miRNA-27a). Based on this phenomenon, a SiND-featured fluorescent sensor has been constructed for “turn-on” detection of the synthetic target miRNA-27a for the first time. This newly developed approach possesses the merits of low cost, simple design, and convenient operation since no enzymatic reaction, toxic reagents, or separation procedures are involved. The established method achieves a detection limit of 0.16?nM, and the relative standard deviation of this method is 9% (1?nM, n ?=?5). The linear range is 0.5–20?nM, and the recoveries in spiked human fluids are in the range of 90–122%. This protocol provides a new tactic in the development of the nonenzymic miRNA biosensors and opens a promising avenue for early diagnosis of miRNA-associated disease. Graphical abstract The SiND-based fluorescent sensor for detection of S-miR-27a.
机译:我们已经开发了一种新的酶序列DNA检测方法,基于荧光硅纳米蛋白(Sinds)朝向Cy5标记的DNA探针的动态猝灭。令人欣然的是,水溶性Sinds可以在均匀溶液中淬灭Cy5标记的DNA探针中的花青(Cy5)的荧光,并且可以在靶序列DNA存在下恢复Cy5标记的DNA探针的荧光(合成靶miRNA -27a)。基于这种现象,首次构建了Sind荧光传感器的荧光传感器,用于“开启”检测合成靶MiRNA-27a的检测。这种新开发的方法具有低成本,简单的设计和操作方便的优点,因为没有酶促反应,有毒试剂或分离程序。建立的方法实现了0.16Ω·Nm的检出限,该方法的相对标准偏差为9%(1?nm,n?=Δ5)。线性范围为0.5-20?nm,尖刺人流体中的回收率在90-122%的范围内。该协议在非酶miRNA生物传感器的开发中提供了新的策略,并开启了对MiRNA相关疾病的早期诊断的有希望的大道。图解摘要基于Sind的荧光传感器,用于检测S-miR-27a。

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    Key Laboratory of Analytical Chemistry for Biology and Medicine (Ministry of Education) College of Chemistry and Molecular Sciences Wuhan University;

    Key Laboratory of Analytical Chemistry for Biology and Medicine (Ministry of Education) College of Chemistry and Molecular Sciences Wuhan University;

    Key Laboratory of Analytical Chemistry for Biology and Medicine (Ministry of Education) College of Chemistry and Molecular Sciences Wuhan University;

    Key Laboratory of Analytical Chemistry for Biology and Medicine (Ministry of Education) College of Chemistry and Molecular Sciences Wuhan University;

    Key Laboratory of Analytical Chemistry for Biology and Medicine (Ministry of Education) College of Chemistry and Molecular Sciences Wuhan University;

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  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类 分析化学;
  • 关键词

    Silicon nanodots; DNA probe; Fluorescent sensor; The synthetic target miRNA-27a;

    机译:硅纳米蛋白;DNA探针;荧光传感器;合成靶miRNA-27a;

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