Dark-Field Microwells toward High-Throughput Direct miRNA Sensing with Gold Nanoparticles
Dark-Field Microwells toward High-Throughput Direct miRNA Sensing with Gold Nanoparticles
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DOI：
10.1021/acssensors.9b00946
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发表时间：
2019-07-01
期刊：
影响因子：
8.9
通讯作者：
Voros, Janos
中科院分区：
文献类型：
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作者：
Hwu, Stephanie;Blickenstorfer, Yves;Voros, Janos
MicroRNA（miRNA）是一类作为理想生物标志物出现的短RNA，因为已发现其表达水平与包括糖尿病和癌症在内的不同类型的疾病相关。miRNA的检测对于早期诊断和疾病监测是非常有益的。然而，由于其小尺寸和低表达水平，miRNA传感仍然很困难。已经开发了诸如定量实时聚合酶链反应（qRT-PCR）、原位杂交和北方印迹的常用技术来定量给定样品中的miRNA。然而，这些方法在护理点实践中面临共同的挑战，因为它们要么需要复杂的样品处理和昂贵的设备，要么灵敏度低。在这里，我们提出了一种基于暗视野微孔的新工具来克服miRNA传感中的这些挑战。这种小型化的设备能够读出金纳米颗粒测定，而不需要暗场显微镜。我们证明了暗视野微孔检测缓冲液和细胞裂解液中miRNA的可行性。暗视野微孔允许以高通量进行负担得起的miRNA感测，这使其成为用于即时诊断的有前途的工具。
MicroRNA (miRNA) is a class of short RNA that is emerging as an ideal biomarker, as its expression level has been found to correlate with different types of diseases including diabetes and cancer. The detection of miRNA is highly beneficial for early diagnostics and disease monitoring. However, miRNA sensing remains difficult because of its small size and low expression levels. Common techniques such as quantitative real-time polymerase chain reaction (qRT-PCR), in situ hybridization and Northern blotting have been developed to quantify miRNA in a given sample. Nevertheless, these methods face common challenges in point-of-care practice as they either require complicated sample handling and expensive equipment, or suffer from low sensitivity. Here we present a new tool based on dark-field microwells to overcome these challenges in miRNA sensing. This miniaturized device enables the readout of a gold nanoparticle assay without the need of a dark-field microscope. We demonstrate the feasibility of the dark-field microwells to detect miRNA in both buffer solution and cell lysate. The dark-field microwells allow affordable miRNA sensing at a high throughput which make them a promising tool for point-of-care diagnostics.