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A simple method for SARS-CoV-2 detection by rRT-PCR without the use of a commercial RNA extraction kit

The World Health Organization (WHO) has declared a pandemic caused by a new coronavirus named SARS-CoV-2. The growing demand for commercial kits used for automated extraction of SARS-CoV-2 RNA, a key step before rRT-PCR diagnosis, could cause a shortage of stocks that hinders the rapid processing of...

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Detalles Bibliográficos
Autores principales: Ulloa, S., Bravo, C., Parra, B., Ramirez, E., Acevedo, A., Fasce, R., Fernandez, J.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier B.V. 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7442555/
https://www.ncbi.nlm.nih.gov/pubmed/32835738
http://dx.doi.org/10.1016/j.jviromet.2020.113960
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author Ulloa, S.
Bravo, C.
Parra, B.
Ramirez, E.
Acevedo, A.
Fasce, R.
Fernandez, J.
author_facet Ulloa, S.
Bravo, C.
Parra, B.
Ramirez, E.
Acevedo, A.
Fasce, R.
Fernandez, J.
author_sort Ulloa, S.
collection PubMed
description The World Health Organization (WHO) has declared a pandemic caused by a new coronavirus named SARS-CoV-2. The growing demand for commercial kits used for automated extraction of SARS-CoV-2 RNA, a key step before rRT-PCR diagnosis, could cause a shortage of stocks that hinders the rapid processing of samples. Although the recommendation is to use automated methods for nucleic acid extraction, alternatives are necessary to replace commercial kits. However, these alternatives should be as reliable as automated methods. This work describes a simple method to detect SARS-CoV-2 from specimens collected in different preservation media. Samples were previously inactivated by heating and precipitating with a PEG/NaCl solution before rRT-PCR assays for Orf1ab, N and S genes. The new method was compared with an automated protocol of nucleic acid extraction. Both procedures showed similar analytical results. Consequently, this simple and inexpensive method is a suitable procedure for laboratory diagnosis of SARS-CoV-2 infection.
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spelling pubmed-74425552020-08-24 A simple method for SARS-CoV-2 detection by rRT-PCR without the use of a commercial RNA extraction kit Ulloa, S. Bravo, C. Parra, B. Ramirez, E. Acevedo, A. Fasce, R. Fernandez, J. J Virol Methods Short Communication The World Health Organization (WHO) has declared a pandemic caused by a new coronavirus named SARS-CoV-2. The growing demand for commercial kits used for automated extraction of SARS-CoV-2 RNA, a key step before rRT-PCR diagnosis, could cause a shortage of stocks that hinders the rapid processing of samples. Although the recommendation is to use automated methods for nucleic acid extraction, alternatives are necessary to replace commercial kits. However, these alternatives should be as reliable as automated methods. This work describes a simple method to detect SARS-CoV-2 from specimens collected in different preservation media. Samples were previously inactivated by heating and precipitating with a PEG/NaCl solution before rRT-PCR assays for Orf1ab, N and S genes. The new method was compared with an automated protocol of nucleic acid extraction. Both procedures showed similar analytical results. Consequently, this simple and inexpensive method is a suitable procedure for laboratory diagnosis of SARS-CoV-2 infection. Elsevier B.V. 2020-11 2020-08-22 /pmc/articles/PMC7442555/ /pubmed/32835738 http://dx.doi.org/10.1016/j.jviromet.2020.113960 Text en © 2020 Elsevier B.V. All rights reserved. Since January 2020 Elsevier has created a COVID-19 resource centre with free information in English and Mandarin on the novel coronavirus COVID-19. The COVID-19 resource centre is hosted on Elsevier Connect, the company's public news and information website. Elsevier hereby grants permission to make all its COVID-19-related research that is available on the COVID-19 resource centre - including this research content - immediately available in PubMed Central and other publicly funded repositories, such as the WHO COVID database with rights for unrestricted research re-use and analyses in any form or by any means with acknowledgement of the original source. These permissions are granted for free by Elsevier for as long as the COVID-19 resource centre remains active.
spellingShingle Short Communication
Ulloa, S.
Bravo, C.
Parra, B.
Ramirez, E.
Acevedo, A.
Fasce, R.
Fernandez, J.
A simple method for SARS-CoV-2 detection by rRT-PCR without the use of a commercial RNA extraction kit
title A simple method for SARS-CoV-2 detection by rRT-PCR without the use of a commercial RNA extraction kit
title_full A simple method for SARS-CoV-2 detection by rRT-PCR without the use of a commercial RNA extraction kit
title_fullStr A simple method for SARS-CoV-2 detection by rRT-PCR without the use of a commercial RNA extraction kit
title_full_unstemmed A simple method for SARS-CoV-2 detection by rRT-PCR without the use of a commercial RNA extraction kit
title_short A simple method for SARS-CoV-2 detection by rRT-PCR without the use of a commercial RNA extraction kit
title_sort simple method for sars-cov-2 detection by rrt-pcr without the use of a commercial rna extraction kit
topic Short Communication
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7442555/
https://www.ncbi.nlm.nih.gov/pubmed/32835738
http://dx.doi.org/10.1016/j.jviromet.2020.113960
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