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Phenol-chloroform-based RNA purification for detection of SARS-CoV-2 by RT-qPCR: Comparison with automated systems
The outbreak of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) rapidly reached pandemic levels. Sufficient testing for SARS-CoV-2 has remained essential for tracking and containing the virus. SARS-CoV-2 testing capabilities are still limited in many countries. Here, we explore the use...
Autores principales: | , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2021
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7904160/ https://www.ncbi.nlm.nih.gov/pubmed/33626071 http://dx.doi.org/10.1371/journal.pone.0247524 |
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author | Dimke, Henrik Larsen, Sanne L. Skov, Marianne N. Larsen, Hanne Hartmeyer, Gitte N. Moeller, Jesper B. |
author_facet | Dimke, Henrik Larsen, Sanne L. Skov, Marianne N. Larsen, Hanne Hartmeyer, Gitte N. Moeller, Jesper B. |
author_sort | Dimke, Henrik |
collection | PubMed |
description | The outbreak of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) rapidly reached pandemic levels. Sufficient testing for SARS-CoV-2 has remained essential for tracking and containing the virus. SARS-CoV-2 testing capabilities are still limited in many countries. Here, we explore the use of conventional RNA purification as an alternative to automated systems for detection of SARS-CoV-2 by RT-qPCR. 87 clinical swab specimens were extracted by conventional phenol-chloroform RNA purification and compared to commercial platforms for RNA extraction and the fully integrated Cobas(®)6800 diagnostic system. Our results show that the conventional RNA extraction is fully comparable to modern automated systems regarding analytical sensitivity and specificity with respect to detection of SARS-CoV-2 as evaluated by RT-qPCR. Moreover, the method is easily scalable and implemented in conventional laboratories as a low cost and suitable alternative to automated systems for the detection of SARS-CoV-2. |
format | Online Article Text |
id | pubmed-7904160 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-79041602021-03-02 Phenol-chloroform-based RNA purification for detection of SARS-CoV-2 by RT-qPCR: Comparison with automated systems Dimke, Henrik Larsen, Sanne L. Skov, Marianne N. Larsen, Hanne Hartmeyer, Gitte N. Moeller, Jesper B. PLoS One Research Article The outbreak of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) rapidly reached pandemic levels. Sufficient testing for SARS-CoV-2 has remained essential for tracking and containing the virus. SARS-CoV-2 testing capabilities are still limited in many countries. Here, we explore the use of conventional RNA purification as an alternative to automated systems for detection of SARS-CoV-2 by RT-qPCR. 87 clinical swab specimens were extracted by conventional phenol-chloroform RNA purification and compared to commercial platforms for RNA extraction and the fully integrated Cobas(®)6800 diagnostic system. Our results show that the conventional RNA extraction is fully comparable to modern automated systems regarding analytical sensitivity and specificity with respect to detection of SARS-CoV-2 as evaluated by RT-qPCR. Moreover, the method is easily scalable and implemented in conventional laboratories as a low cost and suitable alternative to automated systems for the detection of SARS-CoV-2. Public Library of Science 2021-02-24 /pmc/articles/PMC7904160/ /pubmed/33626071 http://dx.doi.org/10.1371/journal.pone.0247524 Text en © 2021 Dimke 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 Dimke, Henrik Larsen, Sanne L. Skov, Marianne N. Larsen, Hanne Hartmeyer, Gitte N. Moeller, Jesper B. Phenol-chloroform-based RNA purification for detection of SARS-CoV-2 by RT-qPCR: Comparison with automated systems |
title | Phenol-chloroform-based RNA purification for detection of SARS-CoV-2 by RT-qPCR: Comparison with automated systems |
title_full | Phenol-chloroform-based RNA purification for detection of SARS-CoV-2 by RT-qPCR: Comparison with automated systems |
title_fullStr | Phenol-chloroform-based RNA purification for detection of SARS-CoV-2 by RT-qPCR: Comparison with automated systems |
title_full_unstemmed | Phenol-chloroform-based RNA purification for detection of SARS-CoV-2 by RT-qPCR: Comparison with automated systems |
title_short | Phenol-chloroform-based RNA purification for detection of SARS-CoV-2 by RT-qPCR: Comparison with automated systems |
title_sort | phenol-chloroform-based rna purification for detection of sars-cov-2 by rt-qpcr: comparison with automated systems |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7904160/ https://www.ncbi.nlm.nih.gov/pubmed/33626071 http://dx.doi.org/10.1371/journal.pone.0247524 |
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