PATHPOD - A loop-mediated isothermal amplification (LAMP)-based point-of-care system for rapid clinical detection of SARS-CoV-2 in hospitals in Denmark

Trieu Nguyen; Aaydha Chidambara Vinayaka; Van Ngoc Huynh; Quyen Than Linh; Sune Zoëga Andreasen; Mohsen Golabi; Dang Duong Bang; Jens Kjølseth Møller; Anders Wolff

doi:10.1016/j.snb.2023.134085

PATHPOD - A loop-mediated isothermal amplification (LAMP)-based point-of-care system for rapid clinical detection of SARS-CoV-2 in hospitals in Denmark

Sens Actuators B Chem. 2023 Oct 1:392:134085. doi: 10.1016/j.snb.2023.134085. Epub 2023 Jun 7.

Authors

Trieu Nguyen¹, Aaydha Chidambara Vinayaka², Van Ngoc Huynh¹, Quyen Than Linh¹, Sune Zoëga Andreasen¹, Mohsen Golabi², Dang Duong Bang², Jens Kjølseth Møller³, Anders Wolff¹

Affiliations

¹ BioLabChip Group, Department of Biotechnology and Biomedicine, Technical University of Denmark, Kongens Lyngby, Denmark.
² Laboratory of Applied Micro and Nanotechnology (LAMINATE), Department of Biotechnology and Biomedicine, Technical University of Denmark, Kongens Lyngby, Denmark.
³ Department of Clinical Microbiology, University Hospital of Southern Denmark, Vejle Hospital, Beriderbakken 4, DK-7100 Vejle, Denmark.

Abstract

Sensitive and rapid detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has been a vital goal in the ongoing COVID-19 pandemic. We present in this comprehensive work, for the first time, detailed fabrication and clinical validation of a point of care (PoC) device for rapid, onsite detection of SARS-CoV-2 using a real-time reverse-transcription loop-mediated isothermal amplification (RT-LAMP) reaction on a polymer cartridge. The PoC system, namely PATHPOD, consisting of a standalone device (weight less than 1.2 kg) and a cartridge, can perform the detection of 10 different samples and two controls in less than 50 min, which is much more rapid than the golden standard real-time reverse-transcription Polymerase Chain Reaction (RT-PCR), typically taking 16-48 h. The novel total internal reflection (TIR) scheme and the reactions inside the cartridge in the PoC device allow monitoring of the diagnostic results in real-time and onsite. The analytical sensitivity and specificity of the PoC test are comparable with the current RT-PCR, with a limit of detection (LOD) down to 30-50 viral genome copies. The robustness of the PATHPOD PoC system has been confirmed by analyzing 398 clinical samples initially examined in two hospitals in Denmark. The clinical sensitivity and specificity of these tests are discussed.

Keywords: COVID-19 pandemic; Clinical sensitivity; Limit of detection (LOD); Point of care (PoC) device; Reverse-transcription, loop-mediated isothermal amplification; SARS-CoV-2.