Loop-mediated isothermal amplification (LAMP) is a nucleic acid amplification technique that rapidly amplifies specific DNA molecules at high yield. In this study, a microfluidic droplet array chip was designed to execute the digital LAMP process. The novel device was capable of 1) creating emulsion droplets, 2) sorting them into a 30 × 8 droplet array, and 3) executing LAMP across the 240 trapped and separated droplets (with a volume of 0.22 nL) after only 40 min of reaction at 56 °C. Nucleic acids were accurately quantified across a dynamic range of 50 to 2.5 × 10³ DNA copies per μL, and the limit of detection was a single DNA molecule. This is the first time that an arrayed emulsion droplet microfluidic device has been used for digital LAMP analysis. When compared to microwell digital nucleic acid amplification assays, this droplet array-based digital LAMP assay eliminates the constraint on the size of the digitized target, which was determined by the dimension of the microwells for its counterparts. Moreover, the capacity for hydrodynamic droplet trapping allows the chip to operate in a one-droplet-to-one-trap manner. This microfluidic chip may therefore become a promising device for digital LAMP-based diagnostics in the near future.

中文翻译：

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一种微流体芯片，能够产生和捕获乳液液滴，用于数字回路介导的等温扩增分析†

环介导的等温扩增（LAMP）是一种核酸扩增技术，可以高产率快速扩增特定的DNA分子。在这项研究中，微流控液滴阵列芯片被设计为执行数字LAMP工艺。这种新颖的设备能够：仅反应40分钟后，就可以：1）生成乳液液滴，2）将其分类为30×8的液滴阵列，以及3）在240个被捕获和分离的液滴（体积为0.22 nL）上执行LAMP。在56°C下。在50到2.5×10 ³的动态范围内准确定量了核酸每μLDNA拷贝数，检测极限是单个DNA分子。这是阵列乳剂微滴微流控设备首次用于数字LAMP分析。当与微孔数字核酸扩增测定法进行比较时，这种基于液滴阵列的数字LAMP测定法消除了对数字化靶标大小的限制，该限制条件是由微孔板对应物的大小决定的。而且，流体动力液滴捕获的能力允许芯片以一滴一对一的方式工作。因此，在不久的将来，这种微流体芯片可能会成为有前途的用于基于LAMP的数字诊断的设备。