An integrated microfluidic platform featuring real-time reverse transcription loop-mediated isothermal amplification for detection of COVID-19

You-Ru Jhou; Chih-Hung Wang; Huey-Pin Tsai; Yan-Shen Shan; Gwo-Bin Lee

doi:10.1016/j.snb.2022.131447

An integrated microfluidic platform featuring real-time reverse transcription loop-mediated isothermal amplification for detection of COVID-19

Sens Actuators B Chem. 2022 May 1:358:131447. doi: 10.1016/j.snb.2022.131447. Epub 2022 Jan 26.

Authors

You-Ru Jhou¹, Chih-Hung Wang¹, Huey-Pin Tsai^{2

3}, Yan-Shen Shan^{4

5}, Gwo-Bin Lee^{1

6}

Affiliations

¹ Department of Power Mechanical Engineering, National Tsing Hua University, Hsinchu, Taiwan.
² Department of Pathology, National Cheng Kung University Hospital, College of Medicine, National Cheng Kung University, Tainan, Taiwan.
³ Department of Medical Laboratory Science and Biotechnology, College of Medicine, National Cheng Kung University, Tainan, Taiwan.
⁴ Institute of Clinical Medicine, National Cheng Kung University Hospital, National Cheng Kung University, Tainan, Taiwan.
⁵ Division of General Surgery, Department of Surgery, National Cheng Kung University Hospital, College of Medicine, National Cheng Kung University, Tainan, Taiwan.
⁶ Institute of Nano Engineering and Microsystems, National Tsing Hua University, Hsinchu, Taiwan.

Abstract

An integrated microfluidic platform (IMP) utilizing real-time reverse-transcription loop-mediated isothermal amplification (RT-LAMP) was developed here for detection and quantification of three genes of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2; i.e., coronavirus diseases 2019 (COVID-19)): RNA-dependent RNA polymerase, the envelope gene, and the nucleocapsid gene for molecular diagnosis. The IMP comprised a microfluidic chip, a temperature control module, a fluidic control module that collectively carried out viral lysis, RNA extraction, RT-LAMP, and the real-time detection within 90 min in an automatic format. A limit of detection of 5 × 10³ copies/reaction for each gene was determined with three samples including synthesized RNAs, inactive viruses, and RNAs extracted from clinical samples; this compact platform could be a useful tool for COVID-19 diagnostics.

Keywords: COVID-19; Loop-mediated isothermal amplification; Microfluidics; Molecular diagnostics; SARS-CoV-2.