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基于分子信标和DNAzyme扩增策略的无酶无标记超灵敏比色法检测Pb(2+)

Enzyme-free and label-free ultra-sensitive colorimetric detection of Pb(2+) using molecular beacon and DNAzyme based amplification strategy.

作者信息

Yun Wen, Cai Dingzhou, Jiang JiaoLai, Zhao Pengxiang, Huang Yu, Sang Ge

机构信息

Science and Technology on Surface Physics and Chemistry Laboratory, P.O. Box No. 9-35, Jiangyou City, Sichuan 621908, PR China.

Chongqing Institute of Green and Intelligent Technology, Chinese Academy of Sciences, No. 266 Fangzheng Avenue, Shuitu Hi-Tech Park, Shuitu Town, Beibei District, Chongqing 400714, PR China.

出版信息

Biosens Bioelectron. 2016 Jun 15;80:187-193. doi: 10.1016/j.bios.2016.01.053. Epub 2016 Jan 22.


DOI:10.1016/j.bios.2016.01.053
PMID:26836648
Abstract

An enzyme-free and label-free colorimetric Pb(2+) sensor based on DNAzyme and molecular beacon (MB) has been developed and demonstrated by recycle using enzyme strand for signal amplification. The substrate strand DNA (S-DNA) of DNAzyme could be converted into MB structure with base pairs of stem part at the both ends. The MB could hybridize with enzyme strand DNA (E-DNA) to form DNAzyme, and be activated and cleaved in the presence of Pb(2+). The cleaved MB is much less stable, releasing from the DNAzyme as two product pieces. The product pieces of MB are flexible and could bind to unmodified AuNPs to effectively stabilize them against salt-induced aggregation. Then, the E-DNA is liberated to catalyze the next reaction and amplify the response signal. By taking advantage of repeated using of E-DNA, our proposed method exhibited high sensitive for Pb(2+) detection in a linear range from 0.05 to 5 nM with detection limit of 20 pM by UV-vis spectrometer. Moreover, this method was also used for determination of Pb(2+) in river water samples with satisfying results. Importantly, this strategy could reach high sensitivity without any modification and complex enzymatic or hairpins based amplification procedures.

摘要

一种基于脱氧核酶和分子信标(MB)的无酶、无标记比色法铅离子(Pb(2+))传感器已被开发出来,并通过循环使用酶链进行信号放大得以证明。脱氧核酶的底物链DNA(S-DNA)可以转化为两端带有茎部碱基对的MB结构。MB可以与酶链DNA(E-DNA)杂交形成脱氧核酶,并在Pb(2+)存在的情况下被激活和切割。切割后的MB稳定性大大降低,作为两个产物片段从脱氧核酶上释放出来。MB的产物片段具有柔韧性,可以与未修饰的金纳米颗粒(AuNPs)结合,有效地稳定它们以防止盐诱导的聚集。然后,E-DNA被释放出来催化下一个反应并放大响应信号。通过利用E-DNA的重复使用,我们提出的方法对Pb(2+)检测具有高灵敏度,在0.05至5 nM的线性范围内,通过紫外可见光谱仪检测限为20 pM。此外,该方法还用于测定河水样品中的Pb(2+),结果令人满意。重要的是,该策略无需任何修饰以及复杂的基于酶或发夹的扩增程序即可达到高灵敏度。

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[1]
Enzyme-free and label-free ultra-sensitive colorimetric detection of Pb(2+) using molecular beacon and DNAzyme based amplification strategy.

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[3]
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[4]
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[5]
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[6]
DNAzyme assisted recycling amplification method for ultrasensitive amperometric determination of lead(II) based on the use of a hairpin assembly on a composite prepared from nitrogen doped graphene, perylenetetracarboxylic anhydride, thionine and gold nanoparticles.

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[7]
Electrochemical Detection of Ultratrace Lead Ion through Attaching and Detaching DNA Aptamer from Electrochemically Reduced Graphene Oxide Electrode.

Nanomaterials (Basel). 2019-5-30

[8]
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[9]
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[10]
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