Department of Mechanical Engineering, University of Maryland, College Park, MD, USA.
Lab Chip. 2016 Sep 21;16(18):3524-31. doi: 10.1039/c6lc00711b. Epub 2016 Jul 27.
A microfluidic platform designed for point-of-care PCR-based nucleic acid diagnostics is described. Compared to established microfluidic PCR technologies, the system is unique in its ability to achieve exceptionally rapid PCR amplification in a low cost thermoplastic format, together with high temperature accuracy enabling effective validation of reaction product by high resolution melt analysis performed in the same chamber as PCR. In addition, the system employs capillary pumping for automated loading of sample into the reaction chamber, combined with an integrated hydrophilic valve for precise self-metering of sample volumes into the device. Using the microfluidic system to target a mutation in the G6PC gene, efficient PCR from human genomic DNA template is achieved with cycle times as low as 14 s, full amplification in 8.5 min, and final melt analysis accurately identifying the desired amplicon.
介绍了一种用于即时护理基于 PCR 的核酸诊断的微流控平台。与已建立的微流控 PCR 技术相比，该系统的独特之处在于能够以低成本热塑性形式实现异常快速的 PCR 扩增，并且具有高精度的温度，可在与 PCR 相同的腔室中进行高分辨率熔解分析，从而有效地验证反应产物。此外，该系统采用毛细管泵送自动将样品加载到反应室中，并结合集成的亲水阀，精确地将样品体积计量到设备中。使用微流控系统针对 G6PC 基因中的突变，从人类基因组 DNA 模板中实现高效的 PCR，循环时间低至 14 秒，8.5 分钟内完全扩增，最终的熔解分析准确识别所需的扩增子。
