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首页> 外文期刊>Analytical chemistry >Sensitive Electrochemical Detection of Telomerase Activity Using Spherical Nucleic Acids Gold Nanoparticles Triggered Mimic-Hybridization Chain Reaction Enzyme-Free Dual Signal Amplification
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Sensitive Electrochemical Detection of Telomerase Activity Using Spherical Nucleic Acids Gold Nanoparticles Triggered Mimic-Hybridization Chain Reaction Enzyme-Free Dual Signal Amplification

机译:使用球形核酸金纳米粒子触发模拟-杂交链反应无酶双信号扩增的敏感电化学检测端粒酶活性。

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摘要

We report an electrochemical sensor for telomerase activity detection based on spherical nucleic acids gold nanoparticles (SNAs AuNPs) triggered mimic-hybridization chain reaction (mimic-HCR) enzyme-free dual signal amplification. In the detection strategy, SNAs AuNPs and two hairpin probes were employed. SNAs AuNPs as the primary amplification element, not only hybridized with the telomeric repeats on the electrode to amplify signal but also initiated the subsequent secondary amplification, mimic-hybridization chain reaction of two hairpin probes. If the cells extracts were positive for telomerase activity, SNAs AuNPs could be captured on the electrode. The carried initiators could trigger an alternative hybridization reaction of two hairpin probes that yielded nicked double helices. The signal was further amplified enzyme-free by numerous hexaammineruthenium(III) chloride ([Ru(NH3)(6)](3+), RuHex) inserting into double-helix DNA long chain by electrostatic interaction, each of which could generate an electrochemical signal at appropriate potential. With this method, a detection limit of down to 2 HeLa cells and a dynamic range of 10-10 000 cells were achieved. Telomerase activities of different cell lines were also successfully evaluated.
机译:我们报告了基于球形核酸金纳米粒子(SNAs AuNPs)触发模拟杂交链反应(mimic-HCR)无酶双重信号放大的端粒酶活性检测的电化学传感器。在检测策略中,采用了SNA AuNPs和两个发夹探针。 SNA AuNPs作为主要的扩增元件,不仅与电极上的端粒重复序列杂交以扩增信号,而且还引发了随后的二次扩增,即两个发夹探针的模拟杂交链反应。如果细胞提取物的端粒酶活性呈阳性,则可以在电极上捕获SNAs AuNPs。携带的引发剂可能触发两个发夹探针的交替杂交反应,从而产生带切口的双螺旋。信号通过无规六氨合氯化钌(III)([Ru(NH3)(6)](3+),RuHex)进一步无酶放大,并通过静电相互作用插入双螺旋DNA长链,每一个均可产生适当电位下的电化学信号。使用此方法,可以实现低至2个HeLa细胞的检测限和10至10 000个细胞的动态范围。还成功评估了不同细胞系的端粒酶活性。

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