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基于纳米氧化铈的 DNA 分析(NanoCeracQ DNA 分析)。

DNA assay based on Nanoceria as Fluorescence Quenchers (NanoCeracQ DNA assay).

机构信息

Department of Chemistry and Biomolecular Science, Clarkson University, Potsdam, New York, 13699, United States.

出版信息

Sci Rep. 2018 Feb 5;8(1):2426. doi: 10.1038/s41598-018-20659-9.

Abstract

Functional nanomaterials with fluorescent or quenching abilities are important for the development of molecular probes for detection and studies of nucleic acids. Here, we describe a new class of molecular nanoprobes, the NanoCeracQ that uses nanoceria particles as a nanoquencher of fluorescent oligonucleotides for rapid and sensitive detection of DNA sequences and hybridization events. We show that nanoceria forms stable and reversible bionanoconjugates with oligonucleotides and can specifically recognize and detect DNA sequences in a single step. In absence of the target DNA, the nanoprobe produced minimal background fluorescence due to the high quenching efficiency of nanoceria. Competitive binding of the target induced a concentration dependent increase in the fluorescence signal due to hybridization and release of the fluorescent tag from the nanoparticle surface. The nanoprobe enabled sensitive detection of the complementary strand with a detection limit of 0.12 nM, using a single step procedure. The results show that biofunctionalized nanoceria can be used as a universal nanoquencher and nanosensing platform for fluorescent DNA detection and studies of nucleic acid interactions. This approach can find broad applications in molecular diagnostics, sensor development, gene expression profiling, imaging and forensic analysis.

摘要

具有荧光或猝灭能力的功能纳米材料对于开发用于检测和研究核酸的分子探针是很重要的。在这里,我们描述了一类新的分子纳米探针,即 NanoCeracQ,它使用纳米氧化铈颗粒作为荧光寡核苷酸的纳米猝灭剂,用于快速和灵敏地检测 DNA 序列和杂交事件。我们表明,纳米氧化铈与寡核苷酸形成稳定且可还原的生物纳米复合物,并且可以特异性地识别和检测单步中的 DNA 序列。在不存在靶 DNA 的情况下,由于纳米氧化铈的高猝灭效率,纳米探针产生最小的背景荧光。由于杂交和从纳米颗粒表面释放荧光标记物,竞争结合靶标诱导荧光信号的浓度依赖性增加。该纳米探针能够通过一步程序以 0.12 nM 的检测限灵敏地检测互补链。结果表明,生物功能化的纳米氧化铈可用作通用的纳米猝灭剂和纳米传感平台,用于荧光 DNA 检测和核酸相互作用的研究。这种方法可以在分子诊断、传感器开发、基因表达谱分析、成像和法医分析等领域得到广泛应用。

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