On-chip screening of SARS-CoV-2 cDNA by LAMP-integrated rotational diffusometry

Dhrubajyoti Das; Cheng-Wen Lin; Han-Sheng Chuang

doi:10.1016/j.talanta.2023.125253

On-chip screening of SARS-CoV-2 cDNA by LAMP-integrated rotational diffusometry

Talanta. 2024 Jan 15:267:125253. doi: 10.1016/j.talanta.2023.125253. Epub 2023 Sep 27.

Authors

Dhrubajyoti Das¹, Cheng-Wen Lin², Han-Sheng Chuang³

Affiliations

¹ Department of Biomedical Engineering, National Cheng Kung University, Tainan, 701, Taiwan.
² Department of Medical Laboratory Science and Biotechnology, China Medical University, Taichung, Taiwan; Department of Medical Laboratory Science and Biotechnology, Asia University, Wufeng, Taichung, 413, Taiwan.
³ Department of Biomedical Engineering, National Cheng Kung University, Tainan, 701, Taiwan; Medical Device Innovation Center, National Cheng Kung University, Tainan, 701, Taiwan. Electronic address: oswaldchuang@mail.ncku.edu.tw.

PMID: 37776805
DOI: 10.1016/j.talanta.2023.125253

Abstract

The unprecedented pandemic has raised the demand for prompt, precise, and large-scale virus detection techniques to control the transmission of contagious illnesses. In this study, a loop-mediated isothermal amplification (LAMP) based on-chip platform was developed to address this challenge using rotational diffusometry and functionalized Janus particles. A recombinant plasmid containing a cDNA sequence of the SARS-CoV-2 non-structural protein 2 (nsp2) gene was employed here as a proof-of-concept for COVID-19 detection. Specifically, designed primers and the functionalized Janus particles were simultaneously loaded on a microfluidic chip to perform the LAMP reaction on a hot plate. The optimal Janus particle concentrations for diffusometric analysis were thoroughly validated, and the performance of the on-chip LAMP reaction was assessed using thermal image analysis. Utilization of the highly sensitive rotational diffusometry achieved a limit of detection of 1 pg/μL in just 10 min with a sample volume of 20 μL. Our method delivered a tenfold higher sensitivity than the conventional method by utilizing only half of its usual required time. Overall, this study proposes a potential nucleic acid (NA) amplification device to aid the rapid diagnosis of various diseases by modifying the primers for different target genes.

Keywords: COVID-19; Janus particles; Loop-mediated isothermal amplification; Microchip; Rotational diffusometry; SARS-CoV-2 cDNA.

MeSH terms

COVID-19 Testing
COVID-19* / diagnosis
DNA Primers
DNA, Complementary
Humans
Nucleic Acid Amplification Techniques / methods
RNA, Viral / analysis
RNA, Viral / genetics
SARS-CoV-2* / genetics
Sensitivity and Specificity

Substances

DNA, Complementary
DNA Primers
RNA, Viral