Smart Medicine >

2024 , Vol. 3 >Issue 2: 20240008

DOI: https://doi.org/10.1002/SMMD.20240008

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

  • Yuan Yuan 1,2 ,
  • Perry Ellis 2 ,
  • Ye Tao 2 ,
  • Dimitri A. Bikos 3,4 ,
  • Emma K. Loveday 3,4 ,
  • Mallory M. Thomas 3,4 ,
  • James N. Wilking 3,4,5 ,
  • Connie B. Chang 3,4,5 ,
  • Fangfu Ye , 1,6 ,
  • David A. Weitz , 2,7,8
Expand
  • 1. Oujiang Laboratory (Zhejiang Lab for Regenerative Medicine, Vision and Brain Health), Wenzhou Institute, University of Chinese Academy of Sciences, Wenzhou, Zhejiang, China
  • 2. John A. Paulson School of Engineering and Applied Sciences, Harvard University, Cambridge, Massachusetts, USA
  • 3. Department of Chemical and Biological Engineering, Montana State University, Bozeman, Montana, USA
  • 4. Center for Biofilm Engineering, Montana State University, Bozeman, Montana, USA
  • 5. Department of Physiology and Biomedical Engineering, Mayo Clinic, Rochester, Minnesota, USA
  • 6. Beijing National Laboratory for Condensed Matter Physics, Institute of Physics, Chinese Academy of Sciences, Beijing, China
  • 7. Department of Physics, Harvard University, Cambridge, Massachusetts, USA
  • 8. Wyss Institute for Biologically Inspired Engineering, Harvard University, Boston, Massachusetts, USA
fye@iphy.ac.cn
weitz@seas.harvard.edu

Received date: 03 Apr 2024

Accepted date: 27 Apr 2024

Copyright

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

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.

Key words： ddRT-LAMP; droplet microfluidics; droplet size; RNA rapid detection; SARS-CoV-2 virus

Cite this article

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[J]. Smart Medicine, 2024 , 3(2) : 20240008 . DOI: 10.1002/SMMD.20240008

Options
Outlines

/

〈 〉