Signal-off Electrochemiluminescence Biosensor Based on Phi29 DNA Polymerase Mediated Strand Displacement Amplification for MicroRNA Detection

被引：147

作者：

Chen, Anyi ^{[1
]}

Gui, Guo-Feng ^{[1
]}

Zhuo, Ying ^{[1
]}

Chai, Ya-Qin ^{[1
]}

Xiang, Yun ^{[1
]}

Yuan, Ruo ^{[1
]}

机构：

[1] Southwest Univ, Coll Chem & Chem Engn, Minist Educ, Key Lab Luminescent & Real Time Analyt Chem, Chongqing 400715, Peoples R China

来源：

ANALYTICAL CHEMISTRY | 2015年 / 87卷 / 12期

关键词：

SENSITIVE DETECTION;

D O I：

10.1021/acs.analchem.5b01168

中图分类号：

O65 [分析化学];

学科分类号：

070302 ; 081704 ;

摘要：

A target induced cycling strand displacement amplification (SDA) mediated by phi29 DNA polymerase (phi29) was first investigated and applied in a signal-off electrochemiluminescence (ECL) biosensor for microRNA (miRNA) detection. Herein, the target miRNA triggered the phi29-mediated SDA which could produce amounts of single-stranded DNA (assistant probe) with accurate and comprehensive nucleotide sequence. Then, the assistant probe hybridized with the capture probe and the ferrocene-labeled probe (Pc-probe) to form a ternary "Y" structure for ECL signal quenching by ferrocene. Therefore, the ECL intensity would decrease with increasing concentration of the target miRNA, and the sensitivity of biosensor would be promoted on account of the efficient signal amplification of the target induced cycling reaction. Besides, a self-enhanced Ru(II) ECL system was designed to obtain a stable and strong initial signal to further improve the sensitivity. The ECL assay for miRNA-21 detection is developed with excellent sensitivity of a concentration variation from 10 aM to 1.0 pM and limit of detection down to 3.3 aM.

引用

页码：6328 / 6334

页数：7

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