Low- and middle-income countries (LMICs) are significantly affected by SARS-CoV-2, partially due to their limited capacity for local production and implementation of molecular testing. Here, we provide detailed methods and validation of a molecular toolkit that can be readily produced and deployed using laboratory equipment available in LMICs. Our results show that lab-scale production of enzymes and nucleic acids can supply over 50,000 tests per production batch. The optimized one-step RT-PCR coupled to CRISPR-Cas12a-mediated detection showed a limit of detection of 102 ge/μL in a turnaround time of 2 h. The clinical validation indicated an overall sensitivity of 80%–88%, while for middle and high viral load samples (Cq ≤ 31) the sensitivity was 92%–100%. The specificity was 96%–100% regardless of viral load. Furthermore, we show that the toolkit can be used with the mobile laboratory Bento Lab, potentially enabling LMICs to implement detection services in unattended remote regions. / Fondo Nacional de Desarrollo Científico, Tecnológico y de Innovación Tecnológica / Revisión por pares
Identifer | oai:union.ndltd.org:PERUUPC/oai:repositorioacademico.upc.edu.pe:10757/658641 |
Date | 22 November 2021 |
Creators | Alcántara, Roberto, Peñaranda, Katherin, Mendoza-Rojas, Gabriel, Nakamoto, Jose A., Martins-Luna, Johanna, del Valle-Mendoza, Juana, Adaui, Vanessa, Milón, Pohl |
Publisher | Cell Press |
Source Sets | Universidad Peruana de Ciencias Aplicadas (UPC) |
Language | English |
Detected Language | English |
Type | info:eu-repo/semantics/article |
Format | application/pdf |
Source | Universidad Peruana de Ciencias Aplicadas (UPC), Repositorio Academico - UPC, Cell Reports Methods, 1, 7 |
Rights | info:eu-repo/semantics/openAccess, Attribution-NonCommercial-ShareAlike 4.0 International, http://creativecommons.org/licenses/by-nc-sa/4.0/ |
Relation | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8529268/ |
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