A one-pot isothermal Fluorogenic Mango II arrays-based assay for label-free detection of miRNA.

IF 5.6 1区化学 Q1 CHEMISTRY, ANALYTICAL Talanta Pub Date : 2024-09-19 DOI:10.1016/j.talanta.2024.126920

Zan Gong, Panpan Yuan, Yuqing Gan, Xi Long, Zhiwei Deng, Yalan Tang, Yanjing Yang, Shian Zhong

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Abstract

The capability to detect a small number of miRNAs in clinical samples with simplicity, selectivity, and sensitivity is immensely valuable, yet it remains a daunting task. Here, we described a novel Mango II aptamers-based sensor for the one-pot, sensitive and specific detection of miRNAs. Target miRNA-initiated mediated catalyzed hairpin assembly (CHA) would allow for the production of plenty of DNA duplexes and the formation of the complete T7 promoter, motivating the rolling circle transcription (RCT). Then, the subsequent RCT process efficiently generates a huge number of repeating RNA Mango II aptamers, brightened by the incorporation of fluorescent dye TO1-B for miRNA quantification, realizing label-free and high signal-to-background ratio. Moreover, this assay possesses a remarkable ability to confer high selectivity, enabling the distinction of miRNAs among family members with mere 1- or 2- nucleotide (nt) differences. By employing the proposed assay, we have successfully achieved a sensitive evaluation of miR-21 content in diverse cell lines and clinical serum samples. This offers a versatile approach for the sensitive assay of miRNA biomarkers in molecular diagnosis.

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基于等温荧光芒果 II 阵列的单次无标记检测 miRNA 方法。

以简单、选择性和灵敏度检测临床样本中少量 miRNA 的能力具有极大的价值，但这仍然是一项艰巨的任务。在这里，我们介绍了一种基于芒果 II 合体的新型传感器，它能对 miRNA 进行一次性、灵敏而特异的检测。由目标 miRNA 引发的催化发夹组装（CHA）可产生大量 DNA 双链并形成完整的 T7 启动子，从而推动滚圆转录（RCT）。随后的 RCT 过程有效地生成了大量重复的 RNA 芒果 II 合体，并通过加入荧光染料 TO1-B 进行 miRNA 定量，实现了无标记和高信噪比。此外，这种检测方法还具有显著的高选择性，能够区分仅有 1 或 2 个核苷酸（nt）差异的 miRNA 家族成员。通过采用所提出的检测方法，我们成功地实现了对不同细胞系和临床血清样本中 miR-21 含量的灵敏评估。这为分子诊断中敏感地检测 miRNA 生物标记物提供了一种多功能方法。

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来源期刊

Talanta 化学-分析化学

CiteScore

12.30

自引率

4.90%

发文量

861

审稿时长

29 days

期刊介绍： Talanta provides a forum for the publication of original research papers, short communications, and critical reviews in all branches of pure and applied analytical chemistry. Papers are evaluated based on established guidelines, including the fundamental nature of the study, scientific novelty, substantial improvement or advantage over existing technology or methods, and demonstrated analytical applicability. Original research papers on fundamental studies, and on novel sensor and instrumentation developments, are encouraged. Novel or improved applications in areas such as clinical and biological chemistry, environmental analysis, geochemistry, materials science and engineering, and analytical platforms for omics development are welcome. Analytical performance of methods should be determined, including interference and matrix effects, and methods should be validated by comparison with a standard method, or analysis of a certified reference material. Simple spiking recoveries may not be sufficient. The developed method should especially comprise information on selectivity, sensitivity, detection limits, accuracy, and reliability. However, applying official validation or robustness studies to a routine method or technique does not necessarily constitute novelty. Proper statistical treatment of the data should be provided. Relevant literature should be cited, including related publications by the authors, and authors should discuss how their proposed methodology compares with previously reported methods.