https://scholars.lib.ntu.edu.tw/handle/123456789/598915
Title: | Hand-Powered Microfluidics for Parallel Droplet Digital Loop-Mediated Isothermal Amplification Assays | Authors: | Yuan H Tian J Chao Y Chien Y.-S Luo R.-H Guo J.-Y Li S Chou Y.-J Shum H.C YI-JU CHOU CHIEN-FU CHEN |
Keywords: | compartmentalization;droplet digital loop-mediated isothermal amplification;droplet generation;droplet microfluidics;multi-sample detection;nucleic acid quantification;pathogen detection;Drops;Emulsification;Fluidic devices;Fluorescence microscopy;Isotherms;Nucleic acids;Open source software;Open systems;Personnel;Detection limits;Droplet generation;Droplet microfluidics;Loop mediated isothermal amplifications;Micro-fluidic devices;Monodispersed droplet;Multiple samples;Pathogen detection;Digital microfluidics;lab on a chip;microfluidics;molecular diagnosis;nucleic acid amplification;Lab-On-A-Chip Devices;Microfluidics;Molecular Diagnostic Techniques;Nucleic Acid Amplification Techniques | Issue Date: | 2021 | Journal Volume: | 6 | Journal Issue: | 8 | Start page/Pages: | 2868-2874 | Source: | ACS Sensors | Abstract: | Droplet digital loop-mediated isothermal amplification (ddLAMP) is an important assay for pathogen detection due to its high accuracy, specificity, and ability to quantify nucleic acids. However, performing ddLAMP requires expensive instrumentation and the need for highly trained personnel with expertise in microfluidics. To make ddLAMP more accessible, a ddLAMP assay is developed, featuring significantly decreased operational difficulty and instrumentation requirements. The proposed assay consists of three simplified steps: (1) droplet generation step, in which a LAMP mixture can be emulsified just by manually pulling a syringe connected to a microfluidic device. In this step, for the first time, we verify that highly monodispersed droplets can be generated with unstable flow rates or pressures, allowing untrained personnel to operate the microfluidic device and perform ddLAMP assay; (2) heating step, in which the droplets are isothermally heated in a water bath, which can be found in most laboratories; and (3) result analysis step, in which the ddLAMP result can be determined using only a fluorescence microscopy and an open-source analyzing software. Throughout the process, no droplet microfluidic expertise or equipment is required. More importantly, the proposed system enables multiple samples to be processed simultaneously with a detection limit of 10 copies/μL. The test is simple and intuitive to operate in most laboratories for multi-sample detection, significantly enhancing the accessibility and detection throughput of the ddLAMP technique. ? 2021 American Chemical Society. |
URI: | https://www.scopus.com/inward/record.uri?eid=2-s2.0-85110260067&doi=10.1021%2facssensors.1c00184&partnerID=40&md5=de148d75afe590e3a31ea2dc5c973a27 https://scholars.lib.ntu.edu.tw/handle/123456789/598915 |
ISSN: | 23793694 | DOI: | 10.1021/acssensors.1c00184 |
Appears in Collections: | 應用力學研究所 |
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