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基于 SERS 的 miR-21 直接和夹心检测方法。

SERS-based direct and sandwich assay methods for mir-21 detection.

机构信息

Department of Food Engineering, Faculty of Engineering, Hacettepe University, Beytepe, 06800 Ankara, Turkey.

出版信息

Analyst. 2014 Mar 7;139(5):1141-7. doi: 10.1039/c3an01600e.

Abstract

In this study, two different assay methods were developed using a surface enhanced Raman scattering (SERS) label for sensitive miR-21 detection. In the first method (direct assay), the miR-21 probe was attached to SERS-labelled, rod-shaped gold nanoparticles and hybridised with the target miR-21, which was previously immobilised onto the gold slide. In the second method (sandwich assay), the target miR-21 was captured by an miR-21 probe immobilised onto the gold slide and hybridised with a second miR-21 probe immobilised on the SERS-labeled, rod-shaped gold nanoparticles. SERS signals of developed assays were obtained via a SERS spectrum of 5,5'-dithiobis(2-nitrobenzoic acid) (DTNB) on the rod-shaped nanoparticles. The calibration curves were plotted to measure the different concentrations of miR-21. The detection limits of the direct and sandwich assays, which last less than 40 min, were found to be 0.36 and 0.85 nM, respectively. The developed SERS-based methods offer rapid, selective, sensitive and easy detection of miR-21, especially compared to conventional PCR-based methods.

摘要

在这项研究中,我们开发了两种使用表面增强拉曼散射(SERS)标记物进行灵敏 miR-21 检测的不同检测方法。在第一种方法(直接检测法)中,将 miR-21 探针连接到 SERS 标记的棒状金纳米粒子上,并与先前固定在金片上的靶 miR-21 杂交。在第二种方法(夹心检测法)中,靶 miR-21 被固定在金片上的 miR-21 探针捕获,并与固定在 SERS 标记的棒状金纳米粒子上的第二 miR-21 探针杂交。通过棒状纳米粒子上的 5,5'-二硫代双(2-硝基苯甲酸)(DTNB)的 SERS 光谱获得开发的检测方法的 SERS 信号。绘制校准曲线以测量不同浓度的 miR-21。直接和夹心检测法的检测限分别为 0.36 和 0.85 nM,检测时间均不到 40 分钟。与传统的基于 PCR 的方法相比,所开发的基于 SERS 的方法提供了快速、选择性、灵敏和易于检测 miR-21 的方法。

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