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首页> 外文期刊>RSC Advances >Single-step and ultrasensitive detection of carcinoembryonic antigen based on an aptamer transduction-mediated exonuclease III-assisted dual-amplification strategy
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Single-step and ultrasensitive detection of carcinoembryonic antigen based on an aptamer transduction-mediated exonuclease III-assisted dual-amplification strategy

机译：基于适体转导介导的核酸外切酶III辅助双扩增策略的一步和超灵敏的检测癌胚抗原

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摘要

Herein, a single-step, rapid and homogenous fluorescence approach for highly sensitive and specific detection of CEA was successfully constructed for the first time using an aptamer binding-induced exonuclease III (Exo III)-mediated dual-amplification strategy. When present, CEA can specifically combine with the aptamer region in H1, resulting in a conformational change of H1 and the exposure of the occluded DNA fragment in the stem regions. Successively, the exposed DNA fragment partially hybridizes with H2 to initiate Exo III-assisted cycling cleavage to release another DNA fragment, which can in turn activate the cycling cleavage of the DNA fluorescence substrate (FS). Therefore, many fluorophore fragments are liberated to produce a significantly amplified fluorescence signal toward CEA detection. By virtue of the Exo III-assisted dual-amplification strategy, this method allows the detection of CEA at the fg mL ~(?1) level with excellent selectivity. Compared with other reported strategies for CEA detection, the Exo III-assisted dual-amplification homogeneous platform only requires a one-step reaction, offering a very simple and low-cost detection. The practical ability of the developed strategy is demonstrated by the detection of CEA in human serum with satisfactory results. Thus, this method shows great potential in assays of many other biological analytes in clinical diagnosis.
机译：在本文中，首次成功地使用适体结合诱导的核酸外切酶III（Exo III）介导的双重扩增策略成功构建了一种用于CEA的高灵敏度和特异性检测的单步，快速，均质的荧光方法。当存在时，CEA可以与H1中的适体区域特异性结合，从而导致H1的构象变化和封闭的DNA片段在茎区域中暴露。随后，暴露的DNA片段与H2部分杂交，以启动Exo III辅助的循环裂解，释放出另一个DNA片段，这又可以激活DNA荧光底物（FS）的循环裂解。因此，许多荧光团片段被释放以产生明显扩增的荧光信号，从而用于CEA检测。借助Exo III辅助的双重扩增策略，该方法可以以fg mL〜（？1）的水平以优异的选择性检测CEA。与其他已报道的CEA检测策略相比，Exo III辅助双放大同质平台仅需一步反应，提供了一种非常简单且低成本的检测方法。通过在人血清中检测CEA证明了所开发策略的实用能力，并获得了令人满意的结果。因此，该方法在临床诊断中许多其他生物分析物的测定中显示出巨大潜力。

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