Jun 04, 2020
A Simple RNA Preparation Method for SARS-CoV-2 detection by RT-qPCR
This collection is a draft, published without a DOI.
- Aniela Wozniak1,
- Catalina Ibarra-Henriquez2,
- Valentina Sebastian3,
- Grace Armijo2,
- Liliana Lamig2,
- Carolina Miranda3,
- Marcela Lagos1,
- Sandra Solari1,
- Ana María Guzmán1,
- Teresa Quiroga1,
- Susan Hitschfeld2,
- Eleodoro Riveras2,
- Marcela Ferres1,
- Rodrigo A. Gutiérrez2,
- Patricia García1,
- Ariel Cerda2
- 1Departamento de Laboratorios Clínicos. Escuela de Medicina. Facultad de Medicina. Pontificia Universidad Católica de chile;
- 2FONDAP Center for Genome Regulation. Millennium Institute for Integrative Biology (iBio), Departamento de Genética Molecular y Microbiología, Pontificia Universidad Católica de Chile, Santiago, 8331150, Chile;
- 3Laboratorio de Microbiología. Servicio de laboratorios Clínicos. Red de Salud UC-CHRISTUS
- Coronavirus Method Development Community
- Reclone.org (The Reagent Collaboration Network)
External link: https://doi.org/10.1101/2020.05.07.083048
Collection Citation: Aniela Wozniak, Catalina Ibarra-Henriquez, Valentina Sebastian, Grace Armijo, Liliana Lamig, Carolina Miranda, Marcela Lagos, Sandra Solari, Ana María Guzmán, Teresa Quiroga, Susan Hitschfeld, Eleodoro Riveras, Marcela Ferres, Rodrigo A. Gutiérrez, Patricia García, Ariel Cerda 2020. A Simple RNA Preparation Method for SARS-CoV-2 detection by RT-qPCR. protocols.io https://protocols.io/view/a-simple-rna-preparation-method-for-sars-cov-2-det-bghpjt5n
License: This is an open access collection distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: May 17, 2020
Last Modified: June 04, 2020
Collection Integer ID: 37135
Keywords: Coronavirus, SARS-CoV-2, RNA extraction,
Abstract
The 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.
Safety warnings
Please refer to the Safety Data Sheets (SDS) for health and environmental hazards. Obtain all necessary approvals from relevant Ethics Committees.
