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基于DSN辅助扩增的胃癌患者血清中miRNA-100的电化学检测

Electrochemical detection of miRNA-100 in the sera of gastric cancer patients based on DSN-assisted amplification.

作者信息

Zhuang Jianjian, Wan Haitao, Zhang Xiaobo

机构信息

College of Life Sciences, Zhejiang University, Hangzhou, 310058, China.

College of Life Sciences, Zhejiang University, Hangzhou, 310058, China.

出版信息

Talanta. 2021 Apr 1;225:121981. doi: 10.1016/j.talanta.2020.121981. Epub 2020 Dec 9.

Abstract

Gastric cancer (GC) is a common malignant digestive tract tumor that leads to high mortality worldwide. Early diagnosis of GC is very important for adequate treatment. However, a rapid, specific and sensitive method for the detection of GC is currently not available. Here, a biosensor CPs/AuNP-AuE, the gold nanoparticle (AuNP)-modified Au electrode (AuE) which was coupled with DNA capture probes (CPs), was developed to detect the content of miR-100 in the sera of GC patients. The results showed that AuNPs were uniformly deposited on the surface of AuE. AuNPs enhanced the electrical conductivity and improved the effective area of AuE. CPs were successfully assembled on AuNP-AuE that could be digested by duplex-specific nuclease (DSN) from the miR-100/CPs complex on the electrode, improving the sensitivity of the biosensor by recycling miR-100. The data revealed that the biosensor was highly specific for the detection of miR-100, which had the ability to distinguish one base-pair mistake in miR-100. The detection of the biosensor for miR-100 ranged from 100 aM to 10 pM and the limit of detection (LOD) was estimated to be 100 aM. The detection results of 100 human sera samples using this biosensor indicated that the cutoff for the detection of gastric cancer was 5 fM. Therefore the biosensor developed in our study served as a rapid, specific and sensitive strategy for the detection of gastric cancer in clinic.

摘要

胃癌(GC)是一种常见的恶性消化道肿瘤,在全球范围内导致高死亡率。GC的早期诊断对于充分治疗非常重要。然而,目前尚无一种快速、特异且灵敏的GC检测方法。在此,我们开发了一种生物传感器CPs/AuNP-AuE,即金纳米颗粒(AuNP)修饰的金电极(AuE)与DNA捕获探针(CPs)相结合,用于检测GC患者血清中miR-100的含量。结果表明,AuNPs均匀沉积在AuE表面。AuNPs增强了AuE的导电性并增大了其有效面积。CPs成功组装在AuNP-AuE上,可被电极上miR-100/CPs复合物中的双链特异性核酸酶(DSN)消化,通过循环利用miR-100提高了生物传感器的灵敏度。数据显示,该生物传感器对miR-100的检测具有高度特异性,能够区分miR-100中一个碱基对的错误。该生物传感器对miR-100的检测范围为100 aM至10 pM,检测限(LOD)估计为100 aM。使用该生物传感器对100份人类血清样本的检测结果表明,胃癌检测的临界值为5 fM。因此,我们研究中开发的生物传感器可作为临床上一种快速、特异且灵敏的胃癌检测策略。

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