Unlocking SARS-CoV-2 detection in low- and middle-income countries

• Autores: Alcántara, Roberto, Peñaranda, Katherin, Mendoza-Rojas, Gabriel, Nakamoto, Jose A., Martins-Luna, Johanna, del Valle-Mendoza, Juana, Adaui, Vanessa, Milón, Pohl
• Formato: artículo
• Fecha de Publicación: 2021
• Institución: Universidad Peruana de Ciencias Aplicadas
• Repositorio: UPC-Institucional
• Lenguaje: inglés
• OAI Identifier: oai:repositorioacademico.upc.edu.pe:10757/658641
• Enlace del recurso: http://hdl.handle.net/10757/658641
• Nivel de acceso: acceso abierto
• Materia: COVID-19; CRISPR; fluorescence; LMICs; molecular testing; RT-PCR; SARS-CoV-2

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.