Fabrication of Polymerase Chain Reaction Plastic Lab-on-a-Chip Device for Rapid Molecular Diagnoses.
Fabrication of Polymerase Chain Reaction Plastic Lab-on-a-Chip Device for Rapid Molecular Diagnoses.
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DOI：
10.5213/inj.1632602.301
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发表时间：
2016-05
影响因子：
2.3
通讯作者：
Lee NY
中科院分区：
文献类型：
--
作者：
Trinh KT;Zhang H;Kang DJ;Kahng SH;Tall BD;Lee NY
我们的目标是制造一种热塑性聚甲基丙烯酸甲酯 (PMMA) 芯片实验室设备，以执行连续流聚合酶链式反应 (PCR)，以快速对食源性病原体细菌进行分子检测。利用聚二甲基硅氧烷界面涂层介导的PMMA基板制造了微型塑料器件，能够在温和的条件下进行粘合，从而避免微通道的变形或塌陷。进行表面表征并测量粘合强度。评估了芯片实验室设备利用实验室制造的便携式双加热器执行芯片 PCR 的可行性。将结果与使用市售热循环仪获得的结果进行比较。 PMMA 芯片实验室设备的设计和制造用于使用食源性病原体作为样品目标进行 PCR。 PMMA 基底之间建立了牢固的结合，这对于在塑料上进行小型化 PCR 至关重要。使用大肠杆菌 O157:H7 和调味品克罗诺杆菌（两种世界范围内的食源性病原体）评估了芯片 PCR 的可行性，并在 25 分钟内成功扩增了目标扩增子。在这项研究中，我们提出了一种低成本、高通量热塑性 PMMA 芯片实验室装置的新颖设计，用于进行微量 PCR，并且使用该装置能够以微小分辨率对两种重要的食源性病原体进行快速分子诊断。在这方面，引入的高度便携的系统设计有可能使许多疾病的PCR调查能够快速准确地进行。
We aim to fabricate a thermoplastic poly(methylmethacrylate) (PMMA) Lab-on-a-Chip device to perform continuous- flow polymerase chain reactions (PCRs) for rapid molecular detection of foodborne pathogen bacteria. A miniaturized plastic device was fabricated by utilizing PMMA substrates mediated by poly(dimethylsiloxane) interfacial coating, enabling bonding under mild conditions, and thus avoiding the deformation or collapse of microchannels. Surface characterizations were carried out and bond strength was measured. The feasibility of the Lab-on-a-Chip device for performing on-chip PCR utilizing a lab-made, portable dual heater was evaluated. The results were compared with those obtained using a commercially available thermal cycler. A PMMA Lab-on-a-Chip device was designed and fabricated for conducting PCR using foodborne pathogens as sample targets. A robust bond was established between the PMMA substrates, which is essential for performing miniaturized PCR on plastic. The feasibility of on-chip PCR was evaluated using Escherichia coli O157:H7 and Cronobacter condimenti, two worldwide foodborne pathogens, and the target amplicons were successfully amplified within 25 minutes. In this study, we present a novel design of a low-cost and high-throughput thermoplastic PMMA Lab-on-a-Chip device for conducting microscale PCR, and we enable rapid molecular diagnoses of two important foodborne pathogens in minute resolution using this device. In this regard, the introduced highly portable system design has the potential to enable PCR investigations of many diseases quickly and accurately.