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

Details

Serval ID
serval:BIB_2D0CDF4FCD77
Type
Article: article from journal or magazin.
Collection
Publications
Title
Impact of cobas PCR Media freezing on SARS-CoV-2 viral RNA integrity and whole genome sequencing analyses.
Journal
Diagnostic microbiology and infectious disease
Author(s)
Benoit P., Point F., Gagnon S., Kaufmann D.E., Tremblay C., Harrigan R.P., Hardy I., Coutlée F., Lapierre S.G.
ISSN
1879-0070 (Electronic)
ISSN-L
0732-8893
Publication state
Published
Issued date
12/2021
Peer-reviewed
Oui
Volume
101
Number
4
Pages
115521
Language
english
Notes
Publication types: Journal Article
Publication Status: ppublish
Abstract
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.
Keywords
COVID-19/diagnosis, COVID-19/virology, COVID-19 Testing/methods, Cryopreservation, Freezing, Genome, Viral, Humans, Molecular Diagnostic Techniques/methods, Nasopharynx/virology, Polymerase Chain Reaction/methods, RNA, Viral/genetics, SARS-CoV-2/genetics, SARS-CoV-2/isolation & purification, Whole Genome Sequencing/methods, Cobas PCR Media, Molecular biology, SARS-CoV-2, Transport Medium, Whole Genome Sequencing
Pubmed
Web of science
Create date
09/05/2023 13:59
Last modification date
29/11/2024 17:53
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