Cascade branch migration-triggered strand displacement amplification for specific and sensitive detection of microRNA

血红素 级联 小RNA 检出限 多重位移放大 临床诊断 化学 分子生物学 生物 聚合酶链反应 色谱法 基因 生物化学 医学 DNA提取 临床心理学 血红素
作者
Yaxing Xie,Yulei Hou,Yang Yu,Jianhong Zhang,Jinyan Long,Mengqi Chen,Xueqing Lang,Xiaolan Yang,Hui Chen
出处
期刊:Analytical Methods [Royal Society of Chemistry]
卷期号:16 (25): 4116-4123
标识
DOI:10.1039/d4ay00765d
摘要

MicroRNAs (miRNAs) have been involved in many biological processes and are regarded as promising biomarkers. The short sequence, low abundance and highly homologous interference sequences greatly hinder the accurate detection of miRNAs. Here, a cascade branch migration-triggered strand displacement amplification (CBM-TSDA) strategy was developed for the first time for specific and sensitive detection of miRNA-155 (miR-155). In the presence of target miR-155, the CBM was initiated and two Y-shaped probes were eventually produced. Next, the Y-shaped probes were transformed into three-way junction (3WJ) structures and triggered the SDA to produce a large number of G-quadruplex (G4) structures. Finally, the increased fluorescence signal of G4/Thioflavin T (ThT) was used to quantify miR-155. Meanwhile, the colorimetric responses of the G4-hemin DNAzyme could be used as supplementary detection to obtain a dual-mode signal readout. This detection strategy showed high detection sensitivity, and the limit of detection was 0.28 pM in the fluorescence detection mode and 0.34 pM in the colorimetric detection mode. Notably, it showed high detection specificity, being able to discriminate the single-base mutations of the target with a high discrimination factor. The strategy also possessed excellent capacity for miR-155 detection in cell lysates and real human blood samples. The developed strategy provides a promising detection platform for miRNA, which may be applied to early clinical diagnosis.
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