Cargando…

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...

Descripción completa

Detalles Bibliográficos
Autores principales: Dimke, Henrik, Larsen, Sanne L., Skov, Marianne N., Larsen, Hanne, Hartmeyer, Gitte N., Moeller, Jesper B.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2021
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
_version_ 1783654872941330432
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
work_keys_str_mv AT dimkehenrik phenolchloroformbasedrnapurificationfordetectionofsarscov2byrtqpcrcomparisonwithautomatedsystems
AT larsensannel phenolchloroformbasedrnapurificationfordetectionofsarscov2byrtqpcrcomparisonwithautomatedsystems
AT skovmariannen phenolchloroformbasedrnapurificationfordetectionofsarscov2byrtqpcrcomparisonwithautomatedsystems
AT larsenhanne phenolchloroformbasedrnapurificationfordetectionofsarscov2byrtqpcrcomparisonwithautomatedsystems
AT hartmeyergitten phenolchloroformbasedrnapurificationfordetectionofsarscov2byrtqpcrcomparisonwithautomatedsystems
AT moellerjesperb phenolchloroformbasedrnapurificationfordetectionofsarscov2byrtqpcrcomparisonwithautomatedsystems