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

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: 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