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基于 δ-FeOOH 和外切酶 III 辅助信号扩增的用于丙型肝炎病毒 DNA 检测的高灵敏和选择性荧光生物传感器。

A highly sensitive and selective fluorescence biosensor for hepatitis C virus DNA detection based on δ-FeOOH and exonuclease III-assisted signal amplification.

机构信息

School of Environment, Henan Key Laboratory for Environmental Pollution Control, Key Laboratory for Yellow River and Huai River Water Environment and Pollution Control, Ministry of Education, Henan Normal University, Xinxiang, Henan, 453007, PR China.

School of Environment, Henan Key Laboratory for Environmental Pollution Control, Key Laboratory for Yellow River and Huai River Water Environment and Pollution Control, Ministry of Education, Henan Normal University, Xinxiang, Henan, 453007, PR China.

出版信息

Talanta. 2020 Mar 1;209:120550. doi: 10.1016/j.talanta.2019.120550. Epub 2019 Nov 14.

Abstract

Developing the high selectivity and sensitivity strategy for nucleic acid detection is crucial for early diagnosis and therapy of diseases. In this work, a novel low back-ground fluorescent sensor platform for the detection of nucleic acid has been developed based on δ-FeOOH nanosheets integrating with exonuclease III-assisted target-recycling signal amplification. Because of the strong binding ability between the single-strand DNA (ssDNA) and the δ-FeOOH nanosheets, the dye-labeled ssDNA probe would be quenched by δ-FeOOH nanosheets through fluorescence resonance energy transfer (FRET). By using magnetic separate properties of δ-FeOOH, the background signal was separated from the sensor system, and the low background sensor system was obtained. After adding the target DNA, a double-strand DNA complex (dsDNA) would be formed between the target DNA and dye-labeled ssDNA probe. Then, the dye-labeled ssDNA probe in the dsDNA complex would be stepwise hydrolyzed into short fragments from 3'-terminus by Exonuclease III, and the fluorescence signal was recovered due to the weak bind affinity between the short fragments and δ-FeOOH nanosheets. By using the fluorescence quenching ability of δ-FeOOH nanosheets and enzyme-assisted target-recycling signal amplification, this strategy could show an excellent selectivity toward hepatitis C virus DNA with a low detection limit of 10 pM. By simply changing the dye-labeled ssDNA probe sequence, this sensing platform can be developed as a universal approach for the simple, sensitive, and selective detection of different target DNA.

摘要

开发高选择性和灵敏度的核酸检测策略对于疾病的早期诊断和治疗至关重要。在这项工作中,我们基于δ-FeOOH 纳米片整合了外切酶 III 辅助的靶标循环信号放大,开发了一种用于核酸检测的新型低背景荧光传感器平台。由于单链 DNA(ssDNA)与 δ-FeOOH 纳米片之间具有很强的结合能力,因此标记染料的 ssDNA 探针会通过荧光共振能量转移(FRET)被 δ-FeOOH 纳米片猝灭。利用 δ-FeOOH 的磁性分离特性,将背景信号与传感器系统分离,从而获得低背景传感器系统。加入目标 DNA 后,目标 DNA 和标记染料的 ssDNA 探针之间会形成双链 DNA 复合物(dsDNA)。然后,外切酶 III 会从 3'-末端逐步将 dsDNA 复合物中的标记染料 ssDNA 探针水解成短片段,由于短片段与 δ-FeOOH 纳米片之间的弱结合亲和力,荧光信号得以恢复。通过利用 δ-FeOOH 纳米片的荧光猝灭能力和酶辅助的靶标循环信号放大,该策略可以对丙型肝炎病毒 DNA 表现出优异的选择性,检测下限低至 10 pM。通过简单地改变标记染料 ssDNA 探针的序列,该传感平台可以作为一种通用方法,用于简单、灵敏和选择性地检测不同的目标 DNA。

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