DSN signal amplification strategy based nanochannels biosensor for the detection of miRNAs

IF 4.8 2区 化学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY
Tang-Bin Liao , Ke-Xin Luo , Ji-Yuan Tu , Yu-Lin Zhang , Guo-Jun Zhang , Zhong-Yue Sun
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Abstract

MiRNA-21 is recognized as an important biological marker for the diagnosis, treatment, and prognosis of breast cancer. Here, we have created a nanochannel biosensor utilizing the duplex-specific nuclease (DSN) signal amplification strategy to achieve the detection of miRNAs. In this system, DNA as the capture probe was covalently immobilized on the surface of nanochannels, which hybridized with the target miRNA and forms RNA/DNA duplexes. DSN could cleave the probe DNA in RNA/DNA duplexes, recycling target miRNA, which may again hybridized with other DNA probes. After N cycles, most of the DNA probes had been cleaved, and the content of miRNA could be quantified by detecting changes in surface charge density. This biosensor can distinguish miR-21 from non-complementary miRNAs and one-base mismatched miRNAs, with reliable detection limits as low as 1 fM in PBS. In addition, we had successfully applied this method to analysis of total RNA samples in MCF-7 cells and HeLa cells, and the nanochannels had also shown excellent responsiveness and strong anti-interference ability. This new method is expected to contribute to miRNA detection in clinical diagnostics, providing a unique approach to detecting and distinguishing disease-associated molecules.

Abstract Image

Abstract Image

基于 DSN 信号放大策略的纳米通道生物传感器用于检测 miRNA。
MiRNA-21 被认为是诊断、治疗和预后乳腺癌的重要生物学标志物。在此,我们利用双链特异性核酸酶(DSN)信号放大策略创建了一种纳米通道生物传感器,以实现对 miRNA 的检测。在该系统中,DNA作为捕获探针被共价固定在纳米通道表面,与目标miRNA杂交,形成RNA/DNA双链。DSN 可裂解 RNA/DNA 双链体中的探针 DNA,回收目标 miRNA,再与其他 DNA 探针杂交。经过 N 个循环后,大部分 DNA 探针都已被裂解,miRNA 的含量可通过检测表面电荷密度的变化来量化。这种生物传感器能将 miR-21 与不互补的 miRNA 和单碱基错配的 miRNA 区分开来,在 PBS 中的检测限低至 1 fM。此外,我们还成功地将这种方法应用于 MCF-7 细胞和 HeLa 细胞中总 RNA 样本的分析,纳米通道也表现出卓越的响应性和很强的抗干扰能力。这种新方法有望为临床诊断中的 miRNA 检测做出贡献,为检测和区分疾病相关分子提供一种独特的方法。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
Bioelectrochemistry
Bioelectrochemistry 生物-电化学
CiteScore
9.10
自引率
6.00%
发文量
238
审稿时长
38 days
期刊介绍: An International Journal Devoted to Electrochemical Aspects of Biology and Biological Aspects of Electrochemistry Bioelectrochemistry is an international journal devoted to electrochemical principles in biology and biological aspects of electrochemistry. It publishes experimental and theoretical papers dealing with the electrochemical aspects of: • Electrified interfaces (electric double layers, adsorption, electron transfer, protein electrochemistry, basic principles of biosensors, biosensor interfaces and bio-nanosensor design and construction. • Electric and magnetic field effects (field-dependent processes, field interactions with molecules, intramolecular field effects, sensory systems for electric and magnetic fields, molecular and cellular mechanisms) • Bioenergetics and signal transduction (energy conversion, photosynthetic and visual membranes) • Biomembranes and model membranes (thermodynamics and mechanics, membrane transport, electroporation, fusion and insertion) • Electrochemical applications in medicine and biotechnology (drug delivery and gene transfer to cells and tissues, iontophoresis, skin electroporation, injury and repair). • Organization and use of arrays in-vitro and in-vivo, including as part of feedback control. • Electrochemical interrogation of biofilms as generated by microorganisms and tissue reaction associated with medical implants.
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