Digital droplet RT-LAMP increases speed of SARS-CoV-2 viral RNA detection

Yuan Yuan, Perry Ellis, Ye Tao, Dimitri A. Bikos, Emma K. Loveday, Mallory M. Thomas, James N. Wilking, Connie B. Chang, Fangfu Ye, David A. Weitz

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Smart Medicine ›› 2024, Vol. 3 ›› Issue (2) : 20240008. DOI: 10.1002/SMMD.20240008
RESEARCH ARTICLE

Digital droplet RT-LAMP increases speed of SARS-CoV-2 viral RNA detection

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Abstract

Nucleic acid amplification testing (NAAT) remains one of the most reliable methods for pathogen identification. However, conventional bulk NAATs may not be sufficiently fast or sensitive enough for the detection of clinically-relevant pathogens in point-of-care testing. Here, we have developed a digital droplet RT-LAMP (ddRT-LAMP) assay that rapidly and quantitatively detects the SARS-CoV-2 viral E gene in microfluidic drops. Droplet partitioning using ddRT-LAMP significantly accelerates detection times across a wide range of template concentrations compared to bulk RT-LAMP assays. We discover that a reduction in droplet diameter decreases assay times up to a certain size, upon which surface adsorption of the RT-LAMP polymerase reduces reaction efficiency. Optimization of drop size and polymerase concentration enables rapid, sensitive, and quantitative detection of the SARS-CoV-2 E gene in only 8 min. These results highlight the potential of ddRT-LAMP assays as an excellent platform for quantitative point-of-care testing.

Keywords

ddRT-LAMP / droplet microfluidics / droplet size / RNA rapid detection / SARS-CoV-2 virus

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Yuan Yuan, Perry Ellis, Ye Tao, Dimitri A. Bikos, Emma K. Loveday, Mallory M. Thomas, James N. Wilking, Connie B. Chang, Fangfu Ye, David A. Weitz. Digital droplet RT-LAMP increases speed of SARS-CoV-2 viral RNA detection. Smart Medicine, 2024, 3(2): 20240008 https://doi.org/10.1002/SMMD.20240008

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2024 2024 The Authors. Smart Medicine published by Wiley-VCH GmbH on behalf of Wenzhou Institute, University of Chinese Academy of Sciences.
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