双金属纳米团簇和双扩增策略：用于miRNA-21检测的超灵敏电化学发光生物传感器。

IF 2.5 4区生物学 Q2 BIOCHEMICAL RESEARCH METHODS

Analytical biochemistry Pub Date : 2025-11-01 Epub Date: 2025-07-26 DOI:10.1016/j.ab.2025.115948

Tong Shen, Hongwei Yu, Xin Xu, Ze Zhang, Huanzhang Yang, Dong Chang

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引用次数: 0

摘要

早期非小细胞肺癌（NSCLC）外泌体中microRNA-21 （miRNA-21）的表达升高为早期诊断提供了重要的希望。然而，检测外泌体miRNA-21由于其低丰度仍然具有挑战性。在这里，我们开发了一种电化学发光（ECL）生物传感器，集成了3D DNA行走器和滚动圈扩增（RCA），用于超灵敏的miRNA-21检测。金-银纳米团簇（AuAgNCs, Au:Ag=4:1）作为ECL发射器，而二铁修饰的DNA （Fc-DNA）猝灭信号（“信号关闭”）。靶miRNA-21激活3D DNA walk，生成RCA引物。RCA产物取代了Fc-DNA，恢复了ECL（“信号开启”）。该生物传感器具有宽线性范围（1 fM-1 nM），检测限为0.14 fM。至关重要的是，它将NSCLC患者（n=10）与健康对照（n=10）和COPD患者（n=10）在临床血清样本中区分开来，差异有统计学意义（P < 0.001）。卓越的稳定性（25天后84%的信号保留）和重复性（RSD=3.95%）进一步支持了其在早期肺癌筛查中的应用。

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Bimetallic nanoclusters and dual-amplification tactic: an ultrasensitive electrochemiluminescence biosensor for miRNA-21 detection.

The elevated expression of microRNA-21 (miRNA-21) in exosomes derived from early-stage non-small cell lung cancer (NSCLC) holds significant promise for early diagnosis. However, detecting exosomal miRNA-21 remains challenging due to its low abundance. Here, we developed an electrochemiluminescence (ECL) biosensor integrating a 3D DNA walker with rolling circle amplification (RCA) for ultrasensitive miRNA-21 detection. Gold-silver nanoclusters (AuAgNCs, Au:Ag = 4:1) served as ECL emitters, while ferrocene-modified DNA (Fc-DNA) quenched the signal ("signal-off"). Target miRNA-21 activated the 3D DNA walker, generating primers for RCA. The RCA products displaced Fc-DNA, restoring ECL ("signal-on"). The biosensor achieved a wide linear range (1 fM-1 nM) with a 0.14 fM detection limit. Critically, it distinguished NSCLC patients (n = 10) from healthy controls (n = 10) and COPD patients (n = 10) with statistical significance (P < 0.001) in clinical serum samples. Excellent stability (84 % signal retention after 25 days) and reproducibility (RSD = 3.95 %) further support its utility for early lung cancer screening.

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来源期刊

Analytical biochemistry 生物-分析化学

CiteScore

5.70

自引率

0.00%

发文量

283

审稿时长

44 days

期刊介绍： The journal''s title Analytical Biochemistry: Methods in the Biological Sciences declares its broad scope: methods for the basic biological sciences that include biochemistry, molecular genetics, cell biology, proteomics, immunology, bioinformatics and wherever the frontiers of research take the field. The emphasis is on methods from the strictly analytical to the more preparative that would include novel approaches to protein purification as well as improvements in cell and organ culture. The actual techniques are equally inclusive ranging from aptamers to zymology. The journal has been particularly active in: -Analytical techniques for biological molecules- Aptamer selection and utilization- Biosensors- Chromatography- Cloning, sequencing and mutagenesis- Electrochemical methods- Electrophoresis- Enzyme characterization methods- Immunological approaches- Mass spectrometry of proteins and nucleic acids- Metabolomics- Nano level techniques- Optical spectroscopy in all its forms. The journal is reluctant to include most drug and strictly clinical studies as there are more suitable publication platforms for these types of papers.