A DNA nanomachine based on rolling circle amplification-bridged two-stage exonuclease III-assisted recycling strategy for label-free multi-amplified biosensing of nucleic acid

文献类型： 外文期刊

第一作者： Xue, Qingwang

作者： Xue, Qingwang;Lv, Yanqin;Cui, Hui;Liu, Jifeng;Gu, Xiaohong;Zhang, Shuqiu;Liu, Jifeng

作者机构：

关键词： Fluorescence detection;G-quadruplex;Exonuclease III-assisted amplification;Rolling circle amplification;Hairpin DNA probe

期刊名称：ANALYTICA CHIMICA ACTA （ 影响因子：6.558； 五年影响因子：6.228 ）

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收录情况： SCI

摘要： An autonomous DNA nanomachine based on rolling circle amplification (RCA)-bridged two-stage exonuclease III (Exo III)-induced recycling amplification (Exo III-RCA-Exo III) was developed for label-free and highly sensitive homogeneous multi-amplified detection of DNA combined with sensitive fluorescence detection technique. According to the configuration, the analysis of DNA is accomplished by recognizing the target to a unlabeled molecular beacon (UMB) that integrates target-binding and signal transducer within one multifunctional design, followed by the target-binding of UMB in duplex DNA removed stepwise by Exo III accompanied by the releasing of target DNA for the successive hybridization and cleavage process and autonomous generation of the primer that initiate RCA process with a rational designed padlock DNA. The RCA products containing thousands of repeated catalytic sequences catalytically hybridize with a hairpin reporter probe that includes a "caged" inactive G-quadruplex sequence (HGP) and were then detected by Exo III-assisted recycling amplification, liberating the active G-quadruplex and generating remarkable ZnPPIX/G-quadruplex fluorescence signals with the help of zinc(II)-protoporphyrin IX (ZnPPIX). The proposed strategy showed a wide dynamic range over 7 orders of magnitude with a low limit of detection of 0.51 aM. In addition, this designed protocol can discriminate mismatched DNA from perfectly matched target DNA, and holds a great potential for early diagnosis in gene-related diseases. (C) 2014 Elsevier B.V. All rights reserved.

分类号： O65

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