Fluorometric detection of mutant DNA oligonucleotide based on toehold strand displacement-driving target recycling strategy and exonuclease III-assisted suppression

被引：45

作者：

Chen, Hong Guo ^{[1
]}

Ren, Wang ^{[1
,2
]}

Jia, Jing ^{[1
]}

Feng, Ji ^{[1
]}

Gao, Zhong Feng ^{[1
]}

Li, Nian Bing ^{[1
]}

Luo, Hong Qun ^{[1
]}

机构：

[1] Southwest Univ, Sch Chem & Chem Engn, Key Lab Ecoenvironm Three Gorges Reservoir Reg, Minist Educ, Chongqing 400715, Peoples R China

[2] Sichuan Univ Sci & Engn, Coll Chem & Pharmaceut Engn, Zigong 643000, Peoples R China

来源：

BIOSENSORS & BIOELECTRONICS | 2016年 / 77卷

基金：

中国国家自然科学基金;

关键词：

Oligonucleotide; Fluorescence; Target recycling; Exonuclease III; SYBR green I; HYBRIDIZATION CHAIN-REACTION; RESONANCE ENERGY-TRANSFER; APTAMER-BASED SENSORS; LABEL-FREE; GRAPHENE OXIDE; ELECTROCHEMICAL DETECTION; POLYDOPAMINE NANOSPHERES; ISOTHERMAL AMPLIFICATION; SENSITIVE DETECTION; AMPLIFIED DETECTION;

D O I：

10.1016/j.bios.2015.09.027

中图分类号：

Q6 [生物物理学];

学科分类号：

071011 ;

摘要：

We describe here a fluorometric assay for sensitive detection of oligonucleotides, based on a target recycling amplification strategy driven by toehold-mediated strand displacement reaction and on exonuclease III (Exo III)-assisted fluorescence background suppression strategy. The network consists of a pair of partially complementary DNA hairpins (HP1 and HP2) with 3' overhang ends, between which the spontaneous hybridization is kinetically hindered by the stems. The target DNA is repeatedly used to trigger a recycling progress between the hairpins, generating numerous HP1-HP2 duplex complexes. Exo III was then employed to digest the double strand parts of the residual hairpins and the intermediate products. The fluorescent dye, SYBR Green I, binds to the double-strand DNA products and emits strong fluorescence to achieve sensitive detection of the target DNA with the detection limit of 5.34 pM. Moreover, this proposed strategy showed high discrimination efficiency towards target DNA against mismatched DNA and was successfully applied in the analysis of human serum sample. (C) 2015 Elsevier B.V. All rights reserved.

引用

页码：40 / 45

页数：6

共 60 条

[21] Dynamic DNA Assemblies Mediated by Binding-Induced DNA Strand Displacement [J].