Department of Biomedical Engineering and Institute for Health Innovation and Technology, National University of Singapore, Singapore 119077, Singapore.
Critical Analytics for Manufacturing Personalized Medicine, Singapore MIT Alliance for Research and Technology, Singapore 138602, Singapore.
ACS Sens. 2024 May 24;9(5):2695-2702. doi: 10.1021/acssensors.4c00750. Epub 2024 May 15.
Quantitative nucleic acid amplification tests are of great importance for diagnostics, but current approaches require complex and costly optical setups that limit their nonlaboratory applications. Herein we describe the implementation of a microfluidics platform that can perform binary DNA-amplification-activated droplet sorting. The digital sort-enabled counting (DISCO) platform enables label-free absolute quantification of the nucleic acid. This is achieved by provoking a pH change in droplets through a loop-mediated isothermal amplification (LAMP) reaction, followed by using sorting by interfacial tension (SIFT) to direct positive and negative droplets to different outlets. With the use of on-chip electrodes at both outlets, we demonstrate that the digital electrical counting of target DNA and RNA can be realized. DISCO is a promising approach for realizing sensitive nucleic acid quantification in point-of-care settings.
定量核酸扩增检测在诊断中具有重要意义，但目前的方法需要复杂且昂贵的光学设备，限制了它们在非实验室环境中的应用。本文描述了一种微流控平台的实现，该平台可以进行二元 DNA 扩增激活的液滴分选。数字分选使能计数 (DISCO) 平台能够对核酸进行无标记的绝对定量。这是通过环介导等温扩增 (LAMP) 反应在液滴中引发 pH 变化来实现的，然后使用界面张力分选 (SIFT) 将阳性和阴性液滴引导到不同的出口。通过在两个出口处使用芯片上电极，我们证明可以实现目标 DNA 和 RNA 的数字电计数。DISCO 是在即时护理环境中实现灵敏核酸定量的一种很有前途的方法。
