Visual detection of miRNAs using enzyme-free amplification reactions and ratiometric fluorescent probes

Zhang, Wenyue; Hao, Wenhui; Liu, Xuetao; Sun, Xiru; Yan, Jingli; Wang, Yucong

doi:10.1016/j.talanta.2020.121332

Cited by 23 publications

(6 citation statements)

References 23 publications

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“…图 14 基于无酶扩增和比率荧光探针检测 miRNA [88] Figure 14 miRNA detection based on enzyme-free amplification reactions and ratiometric fluorescent probes [88] . 我们课题组 [88] 结合 HCR 和 CHA 方法，利用比率荧光探针，建立了 miRNA 的可视化检测方法(图 [97] . circRNA 芯片已成为筛选大量 circRNA 的有力工具.…”

Section: 连接-Pcr 法unclassified

Recent advances of <italic>in vitro</italic> detection methods of RNA biomarkers

Zhang¹,

Yan²,

Cheng³

2021

Sci. Sin.-Chim

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摘要 RNA 生物标志物主要包括编码蛋白的 mRNA 和非编码蛋白的 microRNA (miRNA)、环状 RNA(circular RNA，circRNA)及长链非编码 RNA (lncRNA) 等. RNA 生物标志物种类多，生物信息量丰富，相比 DNA 更能动态反映细胞生物学功能和调控过程，因此，RNA 生物标志物的定量检测和表达分析对于细胞功能研究、疾病的诊断和治疗及预后监测等具有重要意义. 本文重点介绍了常见 RNA 生物标志物 mRNA、miRNA、circRNA 和 lncRNA 体外检测方法的原理及研究进展，并对 RNA 生物标志物检测面临的挑战及发展趋势进行了展望.

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Section: 连接-Pcr 法unclassified

Recent advances of <italic>in vitro</italic> detection methods of RNA biomarkers

Zhang¹,

Yan²,

Cheng³

2021

Sci. Sin.-Chim

Self Cite

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“…For instance, hybridization chain reaction, catalytic hairpin assembly, and cascade toehold-mediated strand displacement reaction (CTSDR). CTSDR shows unique advantages among the above strategies, owing to its flexibility in designing bases [22][23][24][25][26]. Furthermore, it is an enzyme-free reaction with a short time, mild reaction temperatures, and simple operations.…”

Section: Introductionmentioning

confidence: 99%

A strand displacement signal amplification‐assisted fluorescence assay for sensitive detection of microRNA

Chen

Gao

et al. 2023

J of Separation Science

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MicroRNA is a vital biomarker because of its abnormal expression in the emergence and development of diseases, especially in cancers. Herein, a label-free fluorescent sensing platform is proposed for detecting microRNA-21, coupled with the cascade toehold-mediated strand displacement reaction and magnetic beads. Target microRNA-21 acts as an initiator to trigger the cascade toehold-mediated strand displacement reaction and it outputs double-stranded DNA. After magnetic separation, the double-stranded DNA is intercalated by SYBR Green I, resulting in an amplified fluorescent signal. Under the optimal conditions, a wide linear range (0.5-60 nmol/L) and low limits of detection (0.19 nmol/L) are exhibited. What's more, the biosensor shows great specificity and reliability between microRNA-21 and other microRNAs involved in cancer . Owing to the properties of fabulous sensitivity, high selectivity, and simplicity of operator, the proposed method paves a promising way for microRNA-21 detection in cancer diagnosis and biological research.

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“…Effective detection of miRNAs is challenging due to their small size, low expression level and high sequence similarity [ 10 , 11 ]. Recently, several signal amplification strategies, including real-time quantitative polymerase chain reaction (RT-qPCR), hybridization chain reaction, rolling circle amplification, nicking enzyme and duplex-specific nuclease (DSN), have been developed to improve the detection sensitivity for miRNA [ 12 , 13 , 14 , 15 , 16 ]. Further, miRNA biosensing based on DSN-assisted signal amplification is considered a promising strategy.…”

Section: Introductionmentioning

confidence: 99%

Ratiometric Fluorescence Detection of Colorectal Cancer-Associated Exosomal miR-92a-3p with DSN-Assisted Signal Amplification by a MWCNTs@Au NCs Nanoplatform

Sun

Tong

et al. 2022

Biosensors

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The detection of miRNA shows great promise in disease diagnosis. In this work, a ratiometric fluorescent biosensor based on multi-walled carbon nanotubes@gold nanoclusters (MWCNTs@Au NCs) and duplex-specific nuclease (DSN)-assisted signal amplification was fabricated for miRNA detection. Colorectal cancer (CRC)-associated miR-92a-3p extracted from exosomes was selected as the target. MWCNTs@Au NCs performs the dual functions of fluorescence quencher and internal fluorescence reference. In the absence of miR-92a-3p, an Atto-425-modified single-stranded DNA probe is adsorbed on MWCNTs@Au NCs, resulting in the quenching of Atto-425. In the presence of miR-92a-3p, the duplex is formed by hybridization of the probe and miR-92a-3p and leaves the MWCNTs@Au NCs, resulting in the fluorescence recovery of Atto-425. DSN can cleave the probe and result in the release of miR-92a-3p. The released miR-92a-3p can hybridize with other probes to form a signal amplification cycle. The fluorescence of MWCNTs@Au NCs remains stable and constitutes a ratiometric fluorescence system with that of Atto-425. A detection concentration interval of 0.1–10 pM and a limit of detection of 31 fM was obtained under optimized measurement conditions. In addition, the accuracy of the biosensor was validated by detecting the concentration of miR-92a-3p extracted from clinical exosome samples.

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Visual detection of miRNAs using enzyme-free amplification reactions and ratiometric fluorescent probes

Cited by 23 publications

References 23 publications

Recent advances of <italic>in vitro</italic> detection methods of RNA biomarkers

Recent advances of <italic>in vitro</italic> detection methods of RNA biomarkers

A strand displacement signal amplification‐assisted fluorescence assay for sensitive detection of microRNA

Ratiometric Fluorescence Detection of Colorectal Cancer-Associated Exosomal miR-92a-3p with DSN-Assisted Signal Amplification by a MWCNTs@Au NCs Nanoplatform

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Visual detection of miRNAs using enzyme-free amplification reactions and ratiometric fluorescent probes

Cited by 23 publications

References 23 publications

Recent advances of &lt;italic&gt;in vitro&lt;/italic&gt; detection methods of RNA biomarkers

Recent advances of &lt;italic&gt;in vitro&lt;/italic&gt; detection methods of RNA biomarkers

A strand displacement signal amplification‐assisted fluorescence assay for sensitive detection of microRNA

Ratiometric Fluorescence Detection of Colorectal Cancer-Associated Exosomal miR-92a-3p with DSN-Assisted Signal Amplification by a MWCNTs@Au NCs Nanoplatform

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Recent advances of <italic>in vitro</italic> detection methods of RNA biomarkers

Recent advances of <italic>in vitro</italic> detection methods of RNA biomarkers