A Finger-Actuated Sample-Dosing Capillary-Driven Microfluidic Device for Loop-Mediated Isothermal Amplification

Author:

Le Xuan12,Chan Jianxiong1,McMahon James3,Wisniewski Jessica A.3,Coldham Anna3,Alan Tuncay2,Kwan Patrick1

Affiliation:

1. Department of Neuroscience, School of Translational Medicine, Monash University, Melbourne, VIC 3004, Australia

2. Dynamic Micro Devices Laboratory, Department of Mechanical and Aerospace Engineering, Monash University, Clayton, VIC 3800, Australia

3. Department of Infectious Disease, Alfred Hospital and School of Translational Medicine, Monash University, Melbourne, VIC 3004, Australia

Abstract

Loop-mediated isothermal amplification (LAMP) has attracted significant attention for rapid and accurate point-of-care diagnostics. However, integrating sample introduction, lysis, amplification, and detection steps into an easy-to-use, disposable system has so far been challenging. This has limited the uptake of the technique in practical applications. In this study, we developed a colourimetric one-step LAMP assay that combines thermolysis and LAMP reaction, to detect the SARS-CoV-2 virus in nasopharyngeal swab samples from COVID-19-infected individuals. The limit of detection was 500 copies per reaction at 65 °C for 25 min in reaction tubes. Additionally, we developed a finger-operated capillary-driven microfluidic device with selective PVA coating. This finger-actuated microfluidic device could self-dose the required sample amount for the LAMP reaction and inhibit sample evaporation. Finally, we integrated the LAMP assay into the microfluidic device by short-term pre-storage of the LAMP master mix. Using this device, nasopharyngeal swab samples from COVID-19-infected individuals showed positive results at a reaction time of 35 min at 65 °C. This integrated device may be adapted to detect other RNA viruses of interest rapidly.

Funder

NHMRC Investigator Grant

Lord Mayor’s Charitable Foundation and the Ray William Houston Trust

Publisher

MDPI AG

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