Integrated Printed Circuit Board Device for Cell Lysis and Nucleic Acid Extraction
Integrated Printed Circuit Board Device for Cell Lysis and Nucleic Acid Extraction
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DOI：
10.1021/ac302622v
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发表时间：
2012-11-06
影响因子：
7.4
通讯作者：
Santiago, Juan G.
中科院分区：
文献类型：
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作者：
Marshall, Lewis A.;Wu, Liang Li;Santiago, Juan G.
未处理的原始生物样品的制备仍然是微流体的主要挑战。我们提出了一种新的微流控装置的基础上集成的印刷电路板和等速电泳分析样品制备的核酸从生物样品。该装置集成了电阻加热器和温度传感器，以及一个70 μ m × 300 μ m × 31 cm的微流体通道连接两个15 μ L的水库。我们通过从1 μ L掺入恶性疟原虫的全血中提取致病性核酸来证明该装置。我们将全血直接分配到芯片储存器上，系统的集成加热器同时裂解和混合样本。我们使用等速电泳通过等速电泳将核酸提取到二级缓冲液中。我们用微粒子图像测速仪（micro Ply）分析了对流混合作用，并使用芯片外定量聚合酶链反应（PCR）验证了提取的核酸的纯度和量。我们实现了临床相关的检测限为每微升500个寄生虫。该系统没有移动部件，并且该过程可能与广泛的芯片上杂交或扩增测定兼容。
Preparation of raw, untreated biological samples remains a major challenge in microfluidics. We present a novel microfluidic device based on the integration of printed circuit boards and an isotachophoresis assay for sample preparation of nucleic acids from biological samples. The device has integrated resistive heaters and temperature sensors as well as a 70 mu m X 300 mu m X 31 cm microfluidic channel connecting two 15 mu L reservoirs. We demonstrated this device by extracting pathogenic nucleic acids from 1 mu L dispensed volume of whole blood spiked with Plasmodium falciparum. We dispensed whole blood directly onto an on chip reservoir, and the system's integrated heaters simultaneously lysed and mixed the sample. We used isotachophoresis to extract the nucleic acids into a secondary buffer via isotachophoresis. We analyzed the convective mixing action with micro particle image velocimetry (micro Ply) and verified the purity and amount of extracted nucleic acids using off-chip quantitative polymerase chain reaction (PCR). We achieved a clinically relevant limit of detection of 500 parasites per microliter. The system has no moving parts, and the process is potentially compatible with a wide range of on-chip hybridization or amplification assays.