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A catalytic and dual recycling amplification ATP sensor based on target-driven allosteric structure switching of aptamer beacons
Biosensors and Bioelectronics ( IF 12.6 ) Pub Date : 2018-01-09 , DOI: 10.1016/j.bios.2018.01.017
Ying Peng , Daxiu Li , Ruo Yuan , Yun Xiang

Abnormal concentrations of ATP are associated with many diseases and cancers, and quantitative detection of ATP is thus of great importance for disease diagnosis and prognosis. In the present work, we report a new dual recycling amplification sensor integrated with catalytic hairpin assembly (CHA) to achieve high sensitivity for fluorescent detection of ATP. The association of the target ATP with the aptamer beacons causes the allosteric structure switching of the aptamer beacons to expose the toehold regions, which hybridize with and unfold the fluorescently quenched hairpin signal probes (HP1) to recycle the target ATP and to trigger CHA between HP1 and the secondary hairpin probes (HP2) to form HP1/HP2 duplexes. Due to the recycling amplification, the presence of ATP leads to the formation of many HP1/HP2 duplexes, generating dramatically amplified fluorescent signals for sensitive detection of ATP. Under optimal experimental conditions, our sensor linearly responds to ATP in the range from 25 to 600 nM with a calculated detection limit of 8.2 nM. Furthermore, the sensor shows a high selectivity and can also be used to detect ATP in human serums to realize its application for real samples. With the distinct advantage of significant signal amplification without the involvement of any nanomaterial and enzyme, the developed sensor thus holds great potential for simple and sensitive detection of different small molecules and proteins.



中文翻译:

基于适体信标靶点驱动的变构结构转换的催化双循环扩增ATP传感器

ATP浓度异常与许多疾病和癌症有关,因此,ATP的定量检测对于疾病的诊断和预后至关重要。在当前的工作中,我们报告了一种新的双重回收扩增传感器,该传感器与催化发夹组件(CHA)集成在一起,可实现ATP荧光检测的高灵敏度。目标ATP与适体信标的缔合导致适体信标的变构结构转换以暴露脚趾区域,该区域与荧光淬灭的发夹信号探针(HP1)杂交并展开,以回收目标ATP并触发HP1之间的CHA和辅助发夹探针(HP2)形成HP1 / HP2双链体。由于循环扩增,ATP的存在导致许多HP1 / HP2双链体的形成,产生显着放大的荧光信号,以灵敏地检测ATP。在最佳实验条件下,我们的传感器对ATP的线性响应范围为25至600 nM,计算出的检出限为8.2 nM。此外,该传感器具有很高的选择性,还可以用于检测人血清中的ATP,以实现其在实际样品中的应用。凭借显着信号放大而无需任何纳米材料和酶参与的显着优势,开发的传感器因此具有简单而灵敏地检测不同小分子和蛋白质的巨大潜力。该传感器具有很高的选择性,还可以用于检测人血清中的ATP,以实现其在实际样品中的应用。凭借显着信号放大而无需任何纳米材料和酶参与的显着优势,开发的传感器因此具有简单而灵敏地检测不同小分子和蛋白质的巨大潜力。该传感器具有很高的选择性,还可以用于检测人血清中的ATP,以实现其在实际样品中的应用。凭借显着信号放大而无需任何纳米材料和酶参与的显着优势,这种发达的传感器因此具有简单而灵敏地检测不同小分子和蛋白质的巨大潜力。

更新日期:2018-01-09
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