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A simplified SARS-CoV-2 detection protocol for research laboratories

Widespread testing is required to limit the current public health crisis caused by the COVID-19 pandemic. Multiple tests protocols have been authorized by the food and drugs administration (FDA) under an emergency use authorization (EUA). The majority of these protocols are based on the gold-standar...

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Autores principales: Paz, Sean, Mauer, Christopher, Ritchie, Anastasia, Robishaw, Janet D., Caputi, Massimo
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7748277/
https://www.ncbi.nlm.nih.gov/pubmed/33338082
http://dx.doi.org/10.1371/journal.pone.0244271
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author Paz, Sean
Mauer, Christopher
Ritchie, Anastasia
Robishaw, Janet D.
Caputi, Massimo
author_facet Paz, Sean
Mauer, Christopher
Ritchie, Anastasia
Robishaw, Janet D.
Caputi, Massimo
author_sort Paz, Sean
collection PubMed
description Widespread testing is required to limit the current public health crisis caused by the COVID-19 pandemic. Multiple tests protocols have been authorized by the food and drugs administration (FDA) under an emergency use authorization (EUA). The majority of these protocols are based on the gold-standard RT-qPCR test pioneered by the U.S. Centers for Disease Control and Prevention (CDC). However, there is still a widespread lack of testing in the US and many of the clinical diagnostics protocols require extensive human labor and materials that could face supply shortages and present biosafety concerns. Given the need to develop alternative reagents and approaches to provide nucleic-acid testing in the face of heightened demand and potential shortages, we have developed a simplified SARS-CoV-2 testing protocol adapted for its use in research laboratories with minimal molecular biology equipment and expertise. The protocol utilizes TRIzol to purify the viral RNA from different types of clinical specimens, requires minimal BSL-1 precautions and, given its high sensitivity, can be easily adapted to pooling samples strategies.
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spelling pubmed-77482772021-01-07 A simplified SARS-CoV-2 detection protocol for research laboratories Paz, Sean Mauer, Christopher Ritchie, Anastasia Robishaw, Janet D. Caputi, Massimo PLoS One Research Article Widespread testing is required to limit the current public health crisis caused by the COVID-19 pandemic. Multiple tests protocols have been authorized by the food and drugs administration (FDA) under an emergency use authorization (EUA). The majority of these protocols are based on the gold-standard RT-qPCR test pioneered by the U.S. Centers for Disease Control and Prevention (CDC). However, there is still a widespread lack of testing in the US and many of the clinical diagnostics protocols require extensive human labor and materials that could face supply shortages and present biosafety concerns. Given the need to develop alternative reagents and approaches to provide nucleic-acid testing in the face of heightened demand and potential shortages, we have developed a simplified SARS-CoV-2 testing protocol adapted for its use in research laboratories with minimal molecular biology equipment and expertise. The protocol utilizes TRIzol to purify the viral RNA from different types of clinical specimens, requires minimal BSL-1 precautions and, given its high sensitivity, can be easily adapted to pooling samples strategies. Public Library of Science 2020-12-18 /pmc/articles/PMC7748277/ /pubmed/33338082 http://dx.doi.org/10.1371/journal.pone.0244271 Text en © 2020 Paz et al http://creativecommons.org/licenses/by/4.0/ This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle Research Article
Paz, Sean
Mauer, Christopher
Ritchie, Anastasia
Robishaw, Janet D.
Caputi, Massimo
A simplified SARS-CoV-2 detection protocol for research laboratories
title A simplified SARS-CoV-2 detection protocol for research laboratories
title_full A simplified SARS-CoV-2 detection protocol for research laboratories
title_fullStr A simplified SARS-CoV-2 detection protocol for research laboratories
title_full_unstemmed A simplified SARS-CoV-2 detection protocol for research laboratories
title_short A simplified SARS-CoV-2 detection protocol for research laboratories
title_sort simplified sars-cov-2 detection protocol for research laboratories
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7748277/
https://www.ncbi.nlm.nih.gov/pubmed/33338082
http://dx.doi.org/10.1371/journal.pone.0244271
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