External quality assessment of SARS-CoV-2 sequencing: an ESGMD-SSM pilot trial across 15 European laboratories

Wegner, F.; Roloff, T.; Huber, M.; Cordey, S.; Ramette, A.; Gerth, Y.; Bertelli, C.; Stange, M.; Seth-Smith, H.M.B.; Mari, A.; Leuzinger, K.; Cerutti, L.; Harshman, K.; Xenarios, I.; Mercier, P. le; Bittel, P.; Neuenschwander, S.; Opota, O.; Fuchs, J.; Panning, M.; Michel, C.; Hallin, M.; Demuyser, T.; Mendonca, R. de; Savelkoul, P.; Dingemans, J.; Veer, B. van der; Boers, S.A.; Claas, E.C.J.; Coolen, J.P.M.; Melchers, W.J.G.; Gunell, M.; Kallonen, T.; Vuorinen, T.; Hakanen, A.J.; Bernhoff, E.; Hetland, M.A.K.; Berman, H.G.; Adar, S.; Moran-Gilad, J.; Wolf, D.G.; Leib, S.L.; Nolte, O.; Kaiser, L.; Schmutz, S.; Kufner, V.; Zaheri, M.; Trkola, A.; Aamot, H.V.; Hirsch, H.H.; Greub, G.; Egli, A.

doi:doi:10.1128/JCM.01698-21

This first pilot trial on external quality assessment (EQA) of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) whole-genome sequencing, initiated by the European Society of Clinical Microbiology and Infectious Diseases (ESCMID) Study Group for Genomic and Molecular Diagnostics (ESGMD) and the Swiss Society for Microbiology (SSM), aims to build a framework between laboratories in order to improve pathogen surveillance sequencing. Ten samples with various viral loads were sent out to 15 clinical laboratories that had free choice of sequencing methods and bioinformatic analyses. The key aspects on which the individual centers were compared were the identification of (i) single nucleotide polymorphisms (SNPs) and indels, (ii) Pango lineages, and (iii) clusters between samples. The participating laboratories used a wide array of methods and analysis pipelines. Most were able to generate whole genomes for all samples. Genomes were sequenced to various depths (up to a 100...Show moreThis first pilot trial on external quality assessment (EQA) of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) whole-genome sequencing, initiated by the European Society of Clinical Microbiology and Infectious Diseases (ESCMID) Study Group for Genomic and Molecular Diagnostics (ESGMD) and the Swiss Society for Microbiology (SSM), aims to build a framework between laboratories in order to improve pathogen surveillance sequencing. Ten samples with various viral loads were sent out to 15 clinical laboratories that had free choice of sequencing methods and bioinformatic analyses. The key aspects on which the individual centers were compared were the identification of (i) single nucleotide polymorphisms (SNPs) and indels, (ii) Pango lineages, and (iii) clusters between samples. The participating laboratories used a wide array of methods and analysis pipelines. Most were able to generate whole genomes for all samples. Genomes were sequenced to various depths (up to a 100-fold difference across centers). There was a very good consensus regarding the majority of reporting criteria, but there were a few discrepancies in lineage and cluster assignments. Additionally, there were inconsistencies in variant calling. The main reasons for discrepancies were missing data, bioinformatic choices, and interpretation of data. The pilot EQA was overall a success. It was able to show the high quality of participating laboratories and provide valuable feedback in cases where problems occurred, thereby improving the sequencing setup of laboratories. A larger follow-up EQA should, however, improve on defining the variables and format of the report. Additionally, contamination and/or minority variants should be a further aspect of assessment.Show less

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External quality assessment of SARS-CoV-2 sequencing: an ESGMD-SSM pilot trial across 15 European laboratories