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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

Pablo Rodriguez-Mateos

Bongkot Ngamsom

Jesse Gitaka

Alexander Iles

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, Article 338758. 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 Oct 27, 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

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