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A simple RNA preparation method for SARS-CoV-2 detection by RT-qPCR

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 disru...

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Autores principales: 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
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7538882/
https://www.ncbi.nlm.nih.gov/pubmed/33024174
http://dx.doi.org/10.1038/s41598-020-73616-w
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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
author_facet 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
author_sort Wozniak, Aniela
collection PubMed
description 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.
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spelling pubmed-75388822020-10-07 A simple RNA preparation method for SARS-CoV-2 detection by RT-qPCR 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 Sci Rep Article 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. Nature Publishing Group UK 2020-10-06 /pmc/articles/PMC7538882/ /pubmed/33024174 http://dx.doi.org/10.1038/s41598-020-73616-w Text en © The Author(s) 2020 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.
spellingShingle Article
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
A simple RNA preparation method for SARS-CoV-2 detection by RT-qPCR
title A simple RNA preparation method for SARS-CoV-2 detection by RT-qPCR
title_full A simple RNA preparation method for SARS-CoV-2 detection by RT-qPCR
title_fullStr A simple RNA preparation method for SARS-CoV-2 detection by RT-qPCR
title_full_unstemmed A simple RNA preparation method for SARS-CoV-2 detection by RT-qPCR
title_short A simple RNA preparation method for SARS-CoV-2 detection by RT-qPCR
title_sort simple rna preparation method for sars-cov-2 detection by rt-qpcr
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7538882/
https://www.ncbi.nlm.nih.gov/pubmed/33024174
http://dx.doi.org/10.1038/s41598-020-73616-w
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