Author:
Wozniak Aniela,Cerda Ariel,Ibarra-Henríquez Catalina,Sebastian Valentina,Armijo Grace,Lamig Liliana,Miranda Carolina,Lagos Marcela,Solari Sandra,Guzmán Ana María,Quiroga Teresa,Hitschfeld Susan,Riveras Eleodoro,Ferrés Marcela,Gutiérrez Rodrigo A.,García Patricia
Abstract
AbstractThe technique RT-qPCR for viral RNA detection is the current worldwide strategy used for early detection of the novel coronavirus SARS-CoV-2. RNA extraction is a key pre-analytical step in RT-qPCR, often achieved using commercial kits. However, the magnitude of the COVID-19 pandemic is causing disruptions to the global supply chains used by many diagnostic laboratories to procure the commercial kits required for RNA extraction. Shortage in these essential reagents is even more acute in developing countries with no means to produce kits locally. We sought to find an alternative procedure to replace commercial kits using common reagents found in molecular biology laboratories. Here we report a method for RNA extraction that takes about 40 min to complete ten samples, and is not more laborious than current commercial RNA extraction kits. We demonstrate that this method can be used to process nasopharyngeal swab samples and yields RT-qPCR results comparable to those obtained with commercial kits. Most importantly, this procedure can be easily implemented in any molecular diagnostic laboratory. Frequent testing is crucial for individual patient management as well as for public health decision making in this pandemic. Implementation of this method could maintain crucial testing going despite commercial kit shortages.
Funder
Red de Salud UC-CHRISTUS
Department of Clinical Laboratories, School of Medicine, Pontificia Universidad Católica de Chile
Millenium Institute for Integrative Biology – iBio
School of Biological Sciences, Pontificia Universidad Católica de Chile
Fondo de Desarrollo de Áreas Prioritarias (FONDAP) Center for Genome Regulation
Fondo Nacional de Desarrollo Científico y Tecnológico
Publisher
Springer Science and Business Media LLC
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