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Impact of cobas PCR Media freezing on SARS-CoV-2 viral RNA integrity and whole genome sequencing analyses

SARS-CoV-2 whole genome sequencing is a molecular biology tool performed to support many aspects of the response to the pandemic. Freezing of primary clinical nasopharyngeal swabs and shipment to reference laboratories is usually required for sequencing. Cobas PCR Media transport medium facilitates...

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Autores principales: Benoit, Patrick, Point, Floriane, Gagnon, Simon, Kaufmann, Daniel E., Tremblay, Cécile, Harrigan, Richard Paul, Hardy, Isabelle, Coutlée, François, Lapierre, Simon Grandjean
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier Inc. 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8379003/
https://www.ncbi.nlm.nih.gov/pubmed/34537474
http://dx.doi.org/10.1016/j.diagmicrobio.2021.115521
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author Benoit, Patrick
Point, Floriane
Gagnon, Simon
Kaufmann, Daniel E.
Tremblay, Cécile
Harrigan, Richard Paul
Hardy, Isabelle
Coutlée, François
Lapierre, Simon Grandjean
author_facet Benoit, Patrick
Point, Floriane
Gagnon, Simon
Kaufmann, Daniel E.
Tremblay, Cécile
Harrigan, Richard Paul
Hardy, Isabelle
Coutlée, François
Lapierre, Simon Grandjean
author_sort Benoit, Patrick
collection PubMed
description SARS-CoV-2 whole genome sequencing is a molecular biology tool performed to support many aspects of the response to the pandemic. Freezing of primary clinical nasopharyngeal swabs and shipment to reference laboratories is usually required for sequencing. Cobas PCR Media transport medium facilitates high throughput SARS-CoV-2 RT-PCR analyses on cobas platforms. The manufacturer doesn't recommend freezing this transport medium because of risks of degrading molecular templates and impairing test results. Our objective was to compare the quality and results of SARS-CoV-2 genomic sequencing when performed on fresh or frozen samples in cobas PCR Media. Viral genome sequencing was performed using Oxford Nanopore Technologies MinION platform. Sequencing performance, quality and results did not significantly differ between fresh and frozen samples (n = 10). Freezing of cobas PCR Media does not negatively affect SARS-CoV-2 RNA sequencing results and it is therefore a suitable transport medium for outsourcing sequencing analyses to reference laboratories.
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spelling pubmed-83790032021-08-23 Impact of cobas PCR Media freezing on SARS-CoV-2 viral RNA integrity and whole genome sequencing analyses Benoit, Patrick Point, Floriane Gagnon, Simon Kaufmann, Daniel E. Tremblay, Cécile Harrigan, Richard Paul Hardy, Isabelle Coutlée, François Lapierre, Simon Grandjean Diagn Microbiol Infect Dis Article SARS-CoV-2 whole genome sequencing is a molecular biology tool performed to support many aspects of the response to the pandemic. Freezing of primary clinical nasopharyngeal swabs and shipment to reference laboratories is usually required for sequencing. Cobas PCR Media transport medium facilitates high throughput SARS-CoV-2 RT-PCR analyses on cobas platforms. The manufacturer doesn't recommend freezing this transport medium because of risks of degrading molecular templates and impairing test results. Our objective was to compare the quality and results of SARS-CoV-2 genomic sequencing when performed on fresh or frozen samples in cobas PCR Media. Viral genome sequencing was performed using Oxford Nanopore Technologies MinION platform. Sequencing performance, quality and results did not significantly differ between fresh and frozen samples (n = 10). Freezing of cobas PCR Media does not negatively affect SARS-CoV-2 RNA sequencing results and it is therefore a suitable transport medium for outsourcing sequencing analyses to reference laboratories. Elsevier Inc. 2021-12 2021-08-21 /pmc/articles/PMC8379003/ /pubmed/34537474 http://dx.doi.org/10.1016/j.diagmicrobio.2021.115521 Text en © 2021 Elsevier Inc. 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 Article
Benoit, Patrick
Point, Floriane
Gagnon, Simon
Kaufmann, Daniel E.
Tremblay, Cécile
Harrigan, Richard Paul
Hardy, Isabelle
Coutlée, François
Lapierre, Simon Grandjean
Impact of cobas PCR Media freezing on SARS-CoV-2 viral RNA integrity and whole genome sequencing analyses
title Impact of cobas PCR Media freezing on SARS-CoV-2 viral RNA integrity and whole genome sequencing analyses
title_full Impact of cobas PCR Media freezing on SARS-CoV-2 viral RNA integrity and whole genome sequencing analyses
title_fullStr Impact of cobas PCR Media freezing on SARS-CoV-2 viral RNA integrity and whole genome sequencing analyses
title_full_unstemmed Impact of cobas PCR Media freezing on SARS-CoV-2 viral RNA integrity and whole genome sequencing analyses
title_short Impact of cobas PCR Media freezing on SARS-CoV-2 viral RNA integrity and whole genome sequencing analyses
title_sort impact of cobas pcr media freezing on sars-cov-2 viral rna integrity and whole genome sequencing analyses
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8379003/
https://www.ncbi.nlm.nih.gov/pubmed/34537474
http://dx.doi.org/10.1016/j.diagmicrobio.2021.115521
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