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DNAzyme 增强的非线性杂交链式反应用于灵敏检测 microRNA。

A DNAzyme-enhanced nonlinear hybridization chain reaction for sensitive detection of microRNA.

机构信息

Xiangya School of Pharmaceutical Sciences in Central South University, Changsha, Hunan, China.

Hunan Zaochen Nanorobot Co Ltd, Liuyang, Hunan, China.

出版信息

J Biol Chem. 2023 Jun;299(6):104751. doi: 10.1016/j.jbc.2023.104751. Epub 2023 Apr 24.

Abstract

As a typical biomarker, the expression of microRNA is closely related to the occurrence of cancer. However, in recent years, the detection methods have had some limitations in the research and application of microRNAs. In this paper, an autocatalytic platform was constructed through the combination of a nonlinear hybridization chain reaction and DNAzyme to achieve efficient detection of microRNA-21. Fluorescently labeled fuel probes can form branched nanostructures and new DNAzyme under the action of the target, and the newly formed DNAzyme can trigger a new round of reactions, resulting in enhanced fluorescence signals. This platform is a simple, efficient, fast, low-cost, and selective method for the detection of microRNA-21, which can detect microRNA-21 at concentrations as low as 0.004 nM and can distinguish sequence differences by single-base differences. In tissue samples from patients with liver cancer, the platform shows the same detection accuracy as real-time PCR but with better reproducibility. In addition, through the flexible design of the trigger chain, our method could be adapted to detect other nucleic acid biomarkers.

摘要

作为一种典型的生物标志物,microRNA 的表达与癌症的发生密切相关。然而,近年来,在 microRNAs 的研究和应用中,检测方法存在一些局限性。本文通过将非线性杂交链式反应和 DNA zyme 结合起来构建了一个自催化平台,实现了对 microRNA-21 的高效检测。在靶标作用下,荧光标记的燃料探针可以形成分支纳米结构和新的 DNAzyme,新形成的 DNAzyme 可以触发新一轮反应,从而增强荧光信号。该平台是一种简单、高效、快速、低成本、选择性检测 microRNA-21 的方法,能够检测到低至 0.004 nM 的 microRNA-21,并且能够通过单碱基差异区分序列差异。在肝癌患者的组织样本中,该平台与实时 PCR 具有相同的检测准确性,但具有更好的重现性。此外,通过触发链的灵活设计,我们的方法可以适应检测其他核酸生物标志物。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6d89/10220273/303a01f4bf55/gr1.jpg

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