Pablo Rodriguez-Mateos
A lab-on-a-chip platform for integrated extraction and detection of SARS-CoV-2 RNA in resource-limited settings
Rodriguez-Mateos, Pablo; Ngamsom, Bongkot; Walter, Cheryl; Dyer, Charlotte E.; Gitaka, Jesse; Iles, Alexander; Pamme, Nicole
Authors
Bongkot Ngamsom
Dr Cheryl Walter C.Walter@hull.ac.uk
Lecturer in Biomedical Science
Dr Charlotte Dyer C.E.Dyer@hull.ac.uk
Lecturer in Biomedical Sciences
Jesse Gitaka
Dr Alex Iles a.Iles@hull.ac.uk
Experimental Officer - Lab on a Chip
Nicole Pamme
Abstract
Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has caused the unprecedented global pandemic of coronavirus disease-2019 (COVID-19). Efforts are needed to develop rapid and accurate diagnostic tools for extensive testing, allowing for effective containment of the infection via timely identification and isolation of SARS-CoV-2 carriers. Current gold standard nucleic acid tests require many separate steps that need trained personnel to operate specialist instrumentation in laboratory environments, hampering turnaround time and test accessibility, especially in low-resource settings. We devised an integrated on-chip platform coupling RNA extraction based on immiscible filtration assisted by surface tension (IFAST), with RNA amplification and detection via colorimetric reverse-transcription loop mediated isothermal amplification (RT-LAMP), using two sets of primers targeting open reading frame 1a (ORF1a) and nucleoprotein (N) genes of SARS-CoV-2. Results were identified visually, with a colour change from pink to yellow indicating positive amplification, and further confirmed by DNA gel electrophoresis. The specificity of the assay was tested against HCoV-OC43 and H1N1 RNAs. The assay based on use of gene N primers was 100% specific to SARS-CoV-2 with no cross-reactivity to HCoV-OC43 nor H1N1. Proof-of-concept studies on water and artificial sputum containing genomic SARS-CoV-2 RNA showed our IFAST RT-LAMP device to be capable of extracting and detecting 470 SARS-CoV-2 copies mL−1 within 1 h (from sample-in to answer-out). IFAST RT-LAMP is a simple-to-use, integrated, rapid and accurate COVID-19 diagnostic platform, which could provide an attractive means for extensive screening of SARS-CoV-2 infections at point-of-care, especially in resource-constrained settings.
Citation
Rodriguez-Mateos, P., Ngamsom, B., Walter, C., Dyer, C. E., Gitaka, J., Iles, A., & Pamme, N. (2021). A lab-on-a-chip platform for integrated extraction and detection of SARS-CoV-2 RNA in resource-limited settings. Analytica Chimica Acta, 1177, https://doi.org/10.1016/j.aca.2021.338758
Journal Article Type | Article |
---|---|
Acceptance Date | Jun 9, 2021 |
Online Publication Date | Jun 14, 2021 |
Publication Date | Sep 8, 2021 |
Deposit Date | Jun 23, 2021 |
Publicly Available Date | Jun 15, 2022 |
Journal | Analytica Chimica Acta |
Print ISSN | 0003-2670 |
Electronic ISSN | 1873-4324 |
Publisher | Elsevier |
Peer Reviewed | Peer Reviewed |
Volume | 1177 |
Article Number | 338758 |
DOI | https://doi.org/10.1016/j.aca.2021.338758 |
Keywords | COVID-19; SARS-CoV-2; IFAST; LAMP; Point-of-care |
Public URL | https://hull-repository.worktribe.com/output/3792745 |
Files
This file is under embargo until Jun 15, 2022 due to copyright reasons.
Contact N.Pamme@hull.ac.uk to request a copy for personal use.
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