Assessment of SARS-CoV-2 Genome Sequencing: Quality Criteria and Low-Frequency Variants.

Details

Ressource 1Download: JCM.00944-21.pdf (946.96 [Ko])
State: Public
Version: author
License: CC BY 4.0
Serval ID
serval:BIB_6FED6E6FD3E3
Type
Article: article from journal or magazin.
Collection
Publications
Institution
Title
Assessment of SARS-CoV-2 Genome Sequencing: Quality Criteria and Low-Frequency Variants.
Journal
Journal of clinical microbiology
Author(s)
Jacot D., Pillonel T., Greub G., Bertelli C.
ISSN
1098-660X (Electronic)
ISSN-L
0095-1137
Publication state
Published
Issued date
20/09/2021
Peer-reviewed
Oui
Volume
59
Number
10
Pages
e0094421
Language
english
Notes
Publication types: Journal Article
Publication Status: ppublish
Abstract
Although many laboratories worldwide have developed their sequencing capacities in response to the need for SARS-CoV-2 genome-based surveillance of variants, only a few reported some quality criteria to ensure sequence quality before lineage assignment and submission to public databases. Hence, we aimed here to provide simple quality control criteria for SARS-CoV-2 sequencing to prevent erroneous interpretation of low-quality or contaminated data. We retrospectively investigated 647 SARS-CoV-2 genomes obtained over 10 tiled amplicons sequencing runs. We extracted 26 potentially relevant metrics covering the entire workflow from sample selection to bioinformatics analysis. Based on data distribution, critical values were established for 11 selected metrics to prompt further quality investigations for problematic samples, in particular those with a low viral RNA quantity. Low-frequency variants (<70% of supporting reads) can result from PCR amplification errors, sample cross contaminations, or presence of distinct SARS-CoV2 genomes in the sample sequenced. The number and the prevalence of low-frequency variants can be used as a robust quality criterion to identify possible sequencing errors or contaminations. Overall, we propose 11 metrics with fixed cutoff values as a simple tool to evaluate the quality of SARS-CoV-2 genomes, among which are cycle thresholds, mean depth, proportion of genome covered at least 10×, and the number of low-frequency variants combined with mutation prevalence data.
Keywords
COVID-19, Genome, Viral, Humans, RNA, Viral, Retrospective Studies, SARS-CoV-2, accreditation, contamination, genome sequencing, quality assessment, variants of concern
Pubmed
Web of science
Open Access
Yes
Create date
02/08/2021 10:47
Last modification date
21/07/2022 7:10
Usage data