Supramolecular Spectrally Encoded Microgels with Double Strand Probes for Absolute and Direct miRNA Fluorescence Detection at High Sensitivity,Journal of the American Chemical Society

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Supramolecular Spectrally Encoded Microgels with Double Strand Probes for Absolute and Direct miRNA Fluorescence Detection at High Sensitivity
Journal of the American Chemical Society ( IF 14.4 ) Pub Date : 2015-01-28 , DOI: 10.1021/ja511644b
Filippo Causa _{1,

2,

3} , Anna Aliberti ₁ , Angela M. Cusano ₁ , Edmondo Battista ₁ , Paolo A. Netti _{1,

2,

3}

Affiliation

We present novel microgels as a particle-based suspension array for direct and absolute microRNA (miRNA) detection. The microgels feature a flexible molecular architecture, antifouling properties, and enhanced sensitivity with a large dynamic range of detection. Specifically, they possess a core-shell molecular architecture with two different fluorescent dyes for multiplex spectral analyses and are endowed with a fluorescent probe for miRNA detection. Encoding and detection fluorescence signals are distinguishable by nonoverlapping emission spectra. Tunable fluorescence probe conjugation and emission confinement on single microgels allow for ultrasensitive miRNA detection. Indeed, the suspension array has high selectivity and sensitivity with absolute quantification, a detection limit of 10(-15) M, a dynamic range from 10(-9) to 10(-15) M, and higher accuracy than qRT-PCR. The antifouling properties of the microgels also permit the direct measurement of miRNAs in serum, without sample pretreatment or target amplification. A multiplexed assay has been tested for a set of miRNAs chosen as cancer biomarkers.

中文翻译：

具有双链探针的超分子光谱编码微凝胶，用于高灵敏度的绝对和直接 miRNA 荧光检测

我们将新型微凝胶作为基于颗粒的悬浮阵列，用于直接和绝对 microRNA (miRNA) 检测。微凝胶具有灵活的分子结构、防污性能和增强的灵敏度以及大动态检测范围。具体来说，它们具有核壳分子结构，带有两种不同的荧光染料，可用于多重光谱分析，并具有用于 miRNA 检测的荧光探针。编码和检测荧光信号可通过不重叠的发射光谱进行区分。单个微凝胶上的可调荧光探针结合和发射限制允许超灵敏的 miRNA 检测。实际上，悬浮阵列具有绝对定量的高选择性和灵敏度，检测限为 10(-15) M，动态范围从 10(-9) 到 10(-15) M，和比 qRT-PCR 更高的准确度。微凝胶的防污特性还允许直接测量血清中的 miRNA，无需样品预处理或目标扩增。已经对一组被选为癌症生物标志物的 miRNA 进行了多重检测。

更新日期：2015-01-28

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