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基于聚多巴胺纳米球/金纳米簇(AuNC)的纳米平台,用于通过 DNA 酶-I 辅助靶标循环扩增进行多种肿瘤相关 microRNA 的双色同时检测。

Polydopamine Nanosphere/Gold Nanocluster (Au NC)-Based Nanoplatform for Dual Color Simultaneous Detection of Multiple Tumor-Related MicroRNAs with DNase-I-Assisted Target Recycling Amplification.

机构信息

Key Laboratory of Sensor Analysis of Tumor Marker, Ministry of Education, College of Chemistry and Molecular Engineering , Qingdao University of Science and Technology , Qingdao 266042 , P. R. China.

出版信息

Anal Chem. 2018 Mar 20;90(6):4039-4045. doi: 10.1021/acs.analchem.7b05253. Epub 2018 Mar 2.

Abstract

A novel fluorescence resonance energy transfer (FRET)-based platform using polydopamine nanospheres (PDANSs) as energy acceptors and dual colored Au NCs as energy donors for simultaneous detection of multiple tumor-related microRNAs with DNase-I-assisted target recycling amplification was developed for the first time. On the basis of monitoring the change of the recovered fluorescence intensity at 445 and 575 nm upon the addition of targets miRNA-21 and let-7a, these two microRNAs (miRNAs) can be simultaneously quantitatively detected, with detection limits of 4.2 and 3.6 pM (3σ) for miRNA-21 and let-7a, which was almost 20 times lower than that without DNase I. Additionally, semiquantitative determination of miRNA-21 and let-7a can also be realized through photovisualization. Most importantly, serums from normal and breast cancer patients can be visually and directly discriminated without any sample pretreatment by confocal microscope experiments, demonstrating promising potential for auxiliary clinical diagnosis.

摘要

一种新型的荧光共振能量转移(FRET)平台,使用聚多巴胺纳米球(PDANSs)作为能量受体,双色金纳米簇(Au NCs)作为能量供体,用于同时检测多种与肿瘤相关的 microRNAs,采用了 DNA 酶 I 辅助的靶标循环放大反应。首次开发了这种基于监测加入靶标 miRNA-21 和 let-7a 后 445nm 和 575nm 处恢复荧光强度变化的方法,可以同时定量检测这两种 microRNAs(miRNAs),miRNA-21 和 let-7a 的检测限分别为 4.2 和 3.6 pM(3σ),比没有 DNA 酶 I 的情况下低了近 20 倍。此外,还可以通过可视化进行 miRNA-21 和 let-7a 的半定量测定。最重要的是,通过共聚焦显微镜实验,无需任何样本预处理,就可以直接肉眼区分正常人和乳腺癌患者的血清,这表明其在辅助临床诊断方面具有很大的应用潜力。

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