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一种通过外切核酸酶III辅助靶标循环和3D DNA步行器级联扩增的简单、一锅法且超灵敏的DNA传感器。

A simple, one-pot and ultrasensitive DNA sensor via Exo III-Assisted target recycling and 3D DNA walker cascade amplification.

作者信息

Wu Na, Wang Yi-Ting, Wang Xiao-Yan, Chen Xu-Wei, Yang Ting, Wang Jian-Hua

机构信息

Research Center for Analytical Sciences, Department of Chemistry, College of Sciences, Northeastern University, Box 332, Shenyang, 110819, China.

Research Center for Analytical Sciences, Department of Chemistry, College of Sciences, Northeastern University, Box 332, Shenyang, 110819, China.

出版信息

Anal Chim Acta. 2021 Feb 22;1147:15-22. doi: 10.1016/j.aca.2020.12.026. Epub 2020 Dec 21.

Abstract

Rapid, sensitive, and user-friendly nucleic acid detection is of growing importance in early clinical diagnosis. Here, we construct a simple, one-pot and ultrasensitive DNA sensor via exonuclease III (Exo III)-assisted target recycling amplification (ERA) combined with 3D DNA walker cascade amplification. In the presence of single-stranded DNA target, the ERA process is activated to generate numerous walker strands (WS). Thereafter, Exo III-powered WSs autonomously move along magnetic bead (MB)-based 3D track to release numerous AgNCs into the supernatant as an amplified signal output. This biosensor had a low detection limit of 18 fM and an analytical range of 40 fM to 1 pM. Furthermore, the practical application potential of this biosensor was also confirmed by the spiking experiments of p53 into human serum and urine samples.

摘要

快速、灵敏且用户友好的核酸检测在早期临床诊断中日益重要。在此,我们通过核酸外切酶III(Exo III)辅助的靶标循环扩增(ERA)与三维DNA步行器级联扩增相结合,构建了一种简单、一锅法且超灵敏的DNA传感器。在单链DNA靶标存在的情况下,ERA过程被激活以产生大量步行链(WS)。此后,由Exo III驱动的WS沿着基于磁珠(MB)的三维轨道自主移动,将大量银纳米簇(AgNCs)释放到上清液中作为放大的信号输出。这种生物传感器的检测下限低至18 fM,分析范围为40 fM至1 pM。此外,通过将p53加标到人类血清和尿液样本中的实验,也证实了这种生物传感器的实际应用潜力。

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