Department of Biological Systems Engineering, Virginia Tech, Blacksburg, VA, USA.
Department of Mechanical Engineering, University of California, Berkeley, CA, USA.
Lab Chip. 2024 Jul 10;24(14):3490-3497. doi: 10.1039/d4lc00265b.
Point-of-care (POC) diagnostics have emerged as a crucial technology for emerging pathogen detections to enable rapid and on-site detection of infectious diseases. However, current POC devices often suffer from limited sensitivity with poor reliability to provide quantitative readouts. In this paper, we present a self-powered digital loop-mediated isothermal amplification (dLAMP) microfluidic chip (SP-dChip) for the rapid and quantitative detection of nucleic acids. The SP-dChip utilizes a vacuum lung design to passively digitize samples into individual nanoliter wells for high-throughput analysis. The superior digitization scheme is further combined with reverse transcription loop-mediated isothermal amplification (RT-LAMP) to demonstrate dLAMP detection of Zika virus (ZIKV). Firstly, the LAMP assay is loaded into the chip and passively digitized into individual wells. Mineral oil is then pipetted through the chip to differentiate each well as an individual reactor. The chip did not require any external pumping or power input for rapid and reliable results to detect ZIKA RNA as low as 100 copies per μL within one hour. As such, this SP-dChip offers a new class of solutions for truly affordable, portable, and quantitative POC detections for emerging viruses.
即时检测 (POC) 诊断已成为新兴病原体检测的关键技术,能够实现传染病的快速和现场检测。然而,目前的 POC 设备通常存在灵敏度有限和可靠性差的问题,无法提供定量读数。在本文中,我们提出了一种自供电数字环介导等温扩增 (dLAMP) 微流控芯片 (SP-dChip),用于快速定量检测核酸。SP-dChip 利用真空肺设计将样本被动数字化成单个纳升级别的小室,以实现高通量分析。这种优越的数字化方案进一步与逆转录环介导等温扩增 (RT-LAMP) 相结合,展示了 dLAMP 对寨卡病毒 (ZIKV) 的检测。首先,将 LAMP 检测试剂盒加载到芯片中,并将其被动地数字化成单个小室。然后通过芯片吸入矿物油,将每个小室区分成独立的反应室。该芯片无需任何外部泵或电源输入,即可在一小时内快速可靠地检测到低至 100 拷贝/μL 的 ZIKA RNA。因此,这种 SP-dChip 为新兴病毒的真正经济实惠、便携和定量的即时检测提供了一种新的解决方案。