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按需液滴中实时基于发光的双链 DNA 的比色测定。

Real-time luminescence-based colorimetric determination of double-strand DNA in droplet on demand.

机构信息

Key Laboratory of Analytical Chemistry for Biology and Medicine (Ministry of Education), College of Chemistry and Molecular Sciences, Wuhan University, Wuhan 430072, PR China.

出版信息

Biosens Bioelectron. 2012 Feb 15;32(1):43-9. doi: 10.1016/j.bios.2011.11.013. Epub 2011 Dec 8.

DOI:10.1016/j.bios.2011.11.013
PMID:22196878
Abstract

We have developed a new luminescence-based colorimetric droplet platform for the determination of double-stranded DNAs (dsDNA). This colorimetric sensor was realized via choosing a fluorescent ensemble probe comprising water-soluble N-acetylcysteine-capped CdTe quantum dots (QDs) and Ru(bpy)(2)(dppx)(2+) (Ru). To provide a convenient and low cost droplet platform for colorimetry, the microvalve technique was adapted to adjust droplet size precisely, achieve the desired fusion of multiple droplets and trap droplets on demand, as well as implement concentration gradients of DNA on a single chip. In the colorimetric sensor, Ru served as both an effective quencher for QDs and a reporter for dsDNA. With increasing concentration of dsDNA, a gradually enhanced color response was observed because of the competition of dsDNA with QDs for Ru. Under the optimum conditions, this biosensing system exhibited not only good sensitivity and specificity for calf thymus DNA with the detection limit of 1.0 pg, but also coincident performances in diluted human serum with the detection limit of 0.9 pg. The droplet biosensor provides a highly efficient, rapid and visual method for dsDNA analysis. The colorimetric droplet platform could be useful as a simple research tool for the study of limited and precious regents such as protein and virus samples, etc.

摘要

我们开发了一种新的基于发光的比色液滴平台,用于测定双链 DNA(dsDNA)。这种比色传感器是通过选择包含水溶性 N-乙酰半胱氨酸封端的 CdTe 量子点(QD)和 Ru(bpy)(2)(dppx)(2 +)(Ru)的荧光整体探针来实现的。为了提供一个方便且低成本的比色滴液平台,我们采用微阀技术精确调整液滴大小,实现多个液滴的期望融合以及按需捕获液滴,并在单个芯片上实现 DNA 的浓度梯度。在比色传感器中,Ru 既可以有效猝灭 QD,也可以作为 dsDNA 的报告分子。随着 dsDNA 浓度的增加,由于 dsDNA 与 QD 竞争 Ru,观察到逐渐增强的颜色响应。在最佳条件下,该生物传感系统对小牛胸腺 DNA 具有良好的灵敏度和特异性,检测限为 1.0 pg,在稀释的人血清中也具有相同的性能,检测限为 0.9 pg。液滴生物传感器为 dsDNA 分析提供了一种高效、快速和可视化的方法。比色液滴平台可作为一种简单的研究工具,用于研究有限且珍贵的试剂,如蛋白质和病毒样本等。

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