Microfluidic bead trap as a visual bar for quantitative detection of oligonucleotides,Lab on a Chip

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Microfluidic bead trap as a visual bar for quantitative detection of oligonucleotides
Lab on a Chip ( IF 6.1 ) Pub Date : 2017-08-23 00:00:00 , DOI: 10.1039/c7lc00836h
Zichen Zhao _{1,

2,

3} , Yuanye Bao _{1,

2,

3} , Lok Ting Chu _{2,

3,

4} , John Kin Lim Ho _{1,

2,

3} , Ching-Chang Chieng _{1,

2,

3} , Ting-Hsuan Chen _{1,

2,

2,

3,

4}

Affiliation

We demonstrate a microfluidic bead trap capable of forming a dipstick-type bar visible to the naked eye for simple and quantitative detection of oligonucleotides. We use magnetic microparticles (MMPs) and polystyrene microparticles (PMPs) that are connected and form MMPs–targets–PMPs when target oligonucleotides are present, leaving free PMPs with a number inversely proportional to the amount of targets. Using a capillary flow-driven microfluidic circuitry consisting of a magnetic separator to remove the MMPs–targets–PMPs, the free PMPs can be trapped at the narrowing nozzle downstream, forming a visual bar quantifiable based on the length of PMP accumulation. Such a power-free and instrument-free platform enables a limit of detection at 13 fmol (0.65 nM in 20 μl, S/N = 3) of oligonucleotides and is compatible with single-nucleotide polymorphisms and operation in a complex bio-fluid. Moreover, using DNAzyme as the target oligonucleotide that catalyzes a specific hydrolytic cleavage in the presence of lead ions, we demonstrate a model application that detects lead ions with a limit of detection of 12.2 nM (2.5 μg l⁻¹), providing quantitative and visual detection of lead contamination at resource-limited sites.

中文翻译：

微流控珠捕获器作为定量检测寡核苷酸的可视条

我们展示了一种微流控珠捕获器，该捕获器能够形成肉眼可见的量油尺型条形，以简单，定量地检测寡核苷酸。我们使用磁性微粒（MMPs）和聚苯乙烯微粒（PMPs）连接，当存在靶寡核苷酸时形成MMPs-targets-PMP，而游离PMP的数量与靶标的数量成反比。使用由磁分离器组成的毛细管流动驱动的微流体电路去除MMP-目标-PMP，游离的PMP可以被捕获在下游的变窄喷嘴处，从而形成基于PMP累积长度可量化的可视条。这种无动力，无仪器的平台可将检测极限限制为13 fmol（20μl中为0.65 nM，S / N = 3）寡核苷酸，并且与单核苷酸多态性和复杂生物流体中的操作兼容。此外，使用DNAzyme作为靶寡核苷酸，可在铅离子存在下催化特定的水解裂解，我们证明了可检测铅离子的模型应用，其检测限为12.2 nM（2.5μgl^-1），在资源有限的站点提供铅污染的定量和视觉检测。

更新日期：2017-09-04

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