基于核酸外切酶Ⅲ诱导的双重信号放大与MoS<sub>2</sub>纳米片荧光猝灭性质的核酸检测方法

引用本文: 刘宇飞, 薛金涛, 闫慧娟, 杨丽娟, 刘巍, 孙祥德. 基于核酸外切酶Ⅲ诱导的双重信号放大与MoS₂纳米片荧光猝灭性质的核酸检测方法[J]. 分析化学, 2017, 45(3): 303-308. doi: 10.11895/j.issn.0253-3820.160642 shu

Citation: LIU Yu-Fei, XUE Jin-Tao, YAN Hui-Juan, YANG Li-Juan, LIU Wei, SUN Xiang-De. A Dual-signal Amplification Method for DNA Detection Based on Exonuclease Ⅲ and Fluorescence Quenching Ability of MoS₂ Nanosheet[J]. Chinese Journal of Analytical Chemistry, 2017, 45(3): 303-308. doi: 10.11895/j.issn.0253-3820.160642 shu

基于核酸外切酶Ⅲ诱导的双重信号放大与MoS₂纳米片荧光猝灭性质的核酸检测方法

新乡医学院药学院, 新乡 453000
基金项目:
本文系国家自然科学基金项目(No.21505114)和河南省教育厅重点项目(No.14A150010)资助

摘要: 将核酸外切酶Ⅲ诱导的双重信号放大技术与MoS₂纳米片的荧光猝灭性质结合，构建了一种高灵敏高选择性的DNA检测方法。首先设计两条末端修饰荧光基团的探针核酸(HP1和HP2)。由于两条探针核酸具有3'粘性末端，使其不会被核酸外切酶Ⅲ降解，因而被吸附于MoS₂纳米片而猝灭其荧光。当目标DNA存在时，会促使核酸外切酶Ⅲ启动双重信号放大反应，并将探针核酸降解成大量的不能吸附于MoS₂纳米片表面的荧光碎片。在优化条件下，目标DNA浓度在0.5~6.0 pmol/L范围内与荧光信号变化呈良好的线性关系，检出限为0.28 pmol/L。与单重信号放大技术相比，本方法极大改善了分析灵敏度和检出限，且具有良好的单碱基错配区分能力。

关键词:

English

A Dual-signal Amplification Method for DNA Detection Based on Exonuclease Ⅲ and Fluorescence Quenching Ability of MoS₂ Nanosheet

School of Pharmacy, Xinxiang Medical University, Xinxiang 453003, China
Received Date: 29 August 2016
Revised Date: 16 December 2016
Fund Project:
This work was supported by the National Natural Science Foundation of China (No. 21505114) and the Key Project of Henan Provincial Department of Education (No. 14A150010)

Abstract: A highly sensitive and selective DNA biosensor is described based on the fluorescence quenching ability of MoS₂ nanosheet and exonuclease Ⅲ (Exo Ⅲ) assisted dual-signal amplification. In this sensor, the fluorescence probes (HP1 and HP2) cannot be degraded by Exo Ⅲ due to the 3'-termini protrusion and thus are adsorbed on the surface of MoS₂ nanosheets, which will result in the quenching of MoS₂ nanosheets toward the probes and induce a low fluorescent signal. The presence of the target DNA leads to the desorption of probes on the surface of MoS₂ nanosheets due to the hybridization toward probes, generating many fluorescent fragments by Exo Ⅲ digestion and inducing dual-signal amplification. This method can improve the sensitivity and detection limit compared with single amplification method, and shows excellent selectivity in the discrimination of single base mismatched targets. On the basis of the significantly high sensitivity, the developed biosensor can be potentially extended to detect various DNA targets with excellent sequence selectivity.

Key words:

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沈阳化工大学材料科学与工程学院沈阳 110142

基于核酸外切酶Ⅲ诱导的双重信号放大与MoS₂纳米片荧光猝灭性质的核酸检测方法

English

A Dual-signal Amplification Method for DNA Detection Based on Exonuclease Ⅲ and Fluorescence Quenching Ability of MoS₂ Nanosheet

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基于核酸外切酶Ⅲ诱导的双重信号放大与MoS2纳米片荧光猝灭性质的核酸检测方法

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A Dual-signal Amplification Method for DNA Detection Based on Exonuclease Ⅲ and Fluorescence Quenching Ability of MoS2 Nanosheet

CCS Chemistry：嵌段共聚物自组装成 “三明治”二维有机材料，实现纳米级压力传感功能

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