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Comparison of Rapid Nucleic Acid Extraction Methods for SARS-CoV-2 Detection by RT-qPCR

Since 2020, humanity has been facing the COVID-19 pandemic, a respiratory disease caused by the SARS-CoV-2. The world’s response to pandemic went through the development of diagnostics, vaccines and medicines. Regarding diagnostics, an enormous challenge was faced due to shortage of materials to col...

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Autores principales: Silva, Lívia Mara, Riani, Lorena Rodrigues, Silvério, Marcelo Silva, Pereira-Júnior, Olavo dos Santos, Pittella, Frederico
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8946922/
https://www.ncbi.nlm.nih.gov/pubmed/35328154
http://dx.doi.org/10.3390/diagnostics12030601
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author Silva, Lívia Mara
Riani, Lorena Rodrigues
Silvério, Marcelo Silva
Pereira-Júnior, Olavo dos Santos
Pittella, Frederico
author_facet Silva, Lívia Mara
Riani, Lorena Rodrigues
Silvério, Marcelo Silva
Pereira-Júnior, Olavo dos Santos
Pittella, Frederico
author_sort Silva, Lívia Mara
collection PubMed
description Since 2020, humanity has been facing the COVID-19 pandemic, a respiratory disease caused by the SARS-CoV-2. The world’s response to pandemic went through the development of diagnostics, vaccines and medicines. Regarding diagnostics, an enormous challenge was faced due to shortage of materials to collect and process the samples, and to perform reliable mass diagnosis by RT-qPCR. In particular, time-consuming and high cost of nucleic acid extraction procedures have hampered the diagnosis; moreover, several steps in the routine for the preparation of the material makes the extracted sample susceptible to contamination. Here two rapid nucleic acid extraction reagents were compared as extraction procedures for SARS-CoV-2 detection in clinical samples by singleplex and multiplex RT-qPCR analysis, using different transport media, samples with high and low viral load, and different PCR machines. As observed, rapid nucleic acid extraction procedures can be applied for reliable diagnosis using a TaqMan-based assay, over multiple platforms. Ultimately, prompt RNA extraction may reduce costs with reagents and plastics, the chances of contamination, and the overall time to diagnosis by RT-qPCR.
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spelling pubmed-89469222022-03-25 Comparison of Rapid Nucleic Acid Extraction Methods for SARS-CoV-2 Detection by RT-qPCR Silva, Lívia Mara Riani, Lorena Rodrigues Silvério, Marcelo Silva Pereira-Júnior, Olavo dos Santos Pittella, Frederico Diagnostics (Basel) Article Since 2020, humanity has been facing the COVID-19 pandemic, a respiratory disease caused by the SARS-CoV-2. The world’s response to pandemic went through the development of diagnostics, vaccines and medicines. Regarding diagnostics, an enormous challenge was faced due to shortage of materials to collect and process the samples, and to perform reliable mass diagnosis by RT-qPCR. In particular, time-consuming and high cost of nucleic acid extraction procedures have hampered the diagnosis; moreover, several steps in the routine for the preparation of the material makes the extracted sample susceptible to contamination. Here two rapid nucleic acid extraction reagents were compared as extraction procedures for SARS-CoV-2 detection in clinical samples by singleplex and multiplex RT-qPCR analysis, using different transport media, samples with high and low viral load, and different PCR machines. As observed, rapid nucleic acid extraction procedures can be applied for reliable diagnosis using a TaqMan-based assay, over multiple platforms. Ultimately, prompt RNA extraction may reduce costs with reagents and plastics, the chances of contamination, and the overall time to diagnosis by RT-qPCR. MDPI 2022-02-27 /pmc/articles/PMC8946922/ /pubmed/35328154 http://dx.doi.org/10.3390/diagnostics12030601 Text en © 2022 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Silva, Lívia Mara
Riani, Lorena Rodrigues
Silvério, Marcelo Silva
Pereira-Júnior, Olavo dos Santos
Pittella, Frederico
Comparison of Rapid Nucleic Acid Extraction Methods for SARS-CoV-2 Detection by RT-qPCR
title Comparison of Rapid Nucleic Acid Extraction Methods for SARS-CoV-2 Detection by RT-qPCR
title_full Comparison of Rapid Nucleic Acid Extraction Methods for SARS-CoV-2 Detection by RT-qPCR
title_fullStr Comparison of Rapid Nucleic Acid Extraction Methods for SARS-CoV-2 Detection by RT-qPCR
title_full_unstemmed Comparison of Rapid Nucleic Acid Extraction Methods for SARS-CoV-2 Detection by RT-qPCR
title_short Comparison of Rapid Nucleic Acid Extraction Methods for SARS-CoV-2 Detection by RT-qPCR
title_sort comparison of rapid nucleic acid extraction methods for sars-cov-2 detection by rt-qpcr
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8946922/
https://www.ncbi.nlm.nih.gov/pubmed/35328154
http://dx.doi.org/10.3390/diagnostics12030601
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