Interlaboratory Comparison Using Inactivated Authentic Sars-Cov-2 Variants as a Feasible Tool for Quality Control in Covid-19 Wastewater Monitoring

26 Pages Posted: 18 May 2023

See all articles by Alexander Wilhelm

Alexander Wilhelm

Goethe University Frankfurt

Jens Schoth

Emschergenossenschaft/Lippeverban

Christina Meinert-Berning

Ruhrverband

Daniel Bastian

FiW e.V., Aachen, Germany

Helmut Blum

Ludwig Maximilian University of Munich (LMU)

Goffe Elsinga

KWR Water Research Institute

Alexander Graf

Ludwig Maximilian University of Munich (LMU)

Leo Heijnen

KWR Water Research Institute

Johannes Ho

TZW: DVGW-Technologiezentrum Wasser

Mariana Kluge

Technische Universität München (TUM)

Stefan Krebs

Ludwig-Maximilians-Universität München, LAFUGA Genomics, Gene Center

Claudia Stange

TZW: DVGW-Technologiezentrum Wasser

Anna Uchaikina

Technische Universität München (TUM)

Regina Dolny

RWTH Aachen University

Christian Wurzbacher

Technische Universität München (TUM)

Jörg E. Drewes

Technische Universität München (TUM)

Gertjan Medema

KWR Water Research Institute

Andreas Tiehm

TZW: DVGW-Technologiezentrum Wasser

Sandra Ciesek

Goethe University Frankfurt - Institute for Medical Virology

Burkhard Teichgräber

Emschergenossenschaft/Lippeverban

Thomas Wintgens

RWTH Aachen University

Frank-Andreas Weber

FiW e.V., Aachen, Germany

Marek Widera

Goethe University Frankfurt

Abstract

Wastewater-based SARS-CoV-2 epidemiology (WBE) has proven as an excellent tool to monitor pandemic dynamics supporting individual testing strategies. WBE can also be used as an early warning system for monitoring the emergence of novel pathogens or viral variants. However, for a timely transmission of results, sophisticated sample logistics and analytics performed in decentralized laboratories close to the sampling sites are required. Since multiple decentralized laboratories commonly use custom in-house workflows for sample purification and PCR-analysis, comparative quality control of the analytical procedures is essential to report reliable and comparable results.In this study, we performed an interlaboratory comparison at laboratories specialized for PCR and high-throughput-sequencing (HTS)-based WBE analysis. Frozen reserve samples from low COVID-19 incidence periods were spiked with different inactivated authentic SARS-CoV-2 variants in graduated concentrations and ratios. Samples were sent to the participating laboratories for analysis using laboratory specific methods and the reported viral genome copy numbers and the detection of viral variants were compared with the expected values.Despite the different procedures, a high concordance regarding the SARS-CoV-2 PCR quantification could be achieved with low variation between the workflows. PCR-based genotyping was, in dependence of the underlying PCR-assay performance, able to predict the relative amount of variant specific substitutions even in samples with low spike-in amount. The identification of variants by HTS, however, required >100 copies/mL wastewater and had limited predictive value when analyzing at a genome coverage below 60%.This interlaboratory test demonstrates that despite different extraction and analysis methods, a high agreement of the SARS-CoV-2 genome copy equivalents could be achieved. Hence, decentralized SARS-CoV-2 wastewater monitoring is feasible to generate comparable analysis results. However, since not all assays detected the correct variant, prior evaluation of PCR and sequencing workflows as well as sustained quality control such as interlaboratory comparisons are mandatory for correct variant detection.

Note:
Funding Information: This study was supported by the German Federal Ministry of Education and Research (BMBF) funding to the projects COVIDready (grant number 02WRS1621A-D), and partly by the project Abwasser Biomarker CoV-2 (02WRS1557A). This study was in part supported in part by the Goethe-Corona-Fund of the Goethe University Frankfurt (M.W.) and by Health Holland.

Declaration of Interests: The authors report supplies provided by QIAGEN GmbH and analytical equipment provided by Analytik Jena (Endress+Hauser Group) on loan for the duration of the project. Qiagen GmbH and Endress+Hauser are associated industry partner of the COVIDready consortium. The funders had no role in the design of the study, in the collection, analyses, or interpretation of data, in the writing of the manuscript, or in the decision to publish the results. The authors (Alexander Wilhelm and Marek Widera) report supplies provided by QIAGEN GmbH and analytical equipment provided by Analytik Jena (Endress+Hauser Group) for the duration of the project. Claudia Stange, Johannes Ho, and Andreas Tiehm reports financial support was provided by Federal Ministry of Education and Research Bonn Office. Marek Widera reports a relationship with AstraZeneca GmbH that includes: speaking and lecture fees. Sandra Ciesek reports a relationship with BioNTech that includes: clinical advisory board membership. Qiagen GmbH and Endress+Hauser are associated industry partner of the COVIDready consortium. The funders had no role in the design of the study, in the collection, analyses, or interpretation of data, in the writing of the manuscript, or in the decision to publish the results.

Keywords: COVID-19 surveillance, SARS-CoV-2 monitoring, wastewater-based epidemiology (WBE), variant detection, interlaboratory comparison, genome sequencing

Suggested Citation

Wilhelm, Alexander and Schoth, Jens and Meinert-Berning, Christina and Bastian, Daniel and Blum, Helmut and Elsinga, Goffe and Graf, Alexander and Heijnen, Leo and Ho, Johannes and Kluge, Mariana and Krebs, Stefan and Stange, Claudia and Uchaikina, Anna and Dolny, Regina and Wurzbacher, Christian and Drewes, Jörg E. and Medema, Gertjan and Tiehm, Andreas and Ciesek, Sandra and Teichgräber, Burkhard and Wintgens, Thomas and Weber, Frank-Andreas and Widera, Marek, Interlaboratory Comparison Using Inactivated Authentic Sars-Cov-2 Variants as a Feasible Tool for Quality Control in Covid-19 Wastewater Monitoring. Available at SSRN: https://ssrn.com/abstract=4442612 or http://dx.doi.org/10.2139/ssrn.4442612

Alexander Wilhelm

Goethe University Frankfurt ( email )

Jens Schoth

Emschergenossenschaft/Lippeverban ( email )

Daniel Bastian

FiW e.V., Aachen, Germany ( email )

Helmut Blum

Ludwig Maximilian University of Munich (LMU) ( email )

Geschwister-Scholl-Platz 1
Munich, DE Bavaria 80539
Germany

Goffe Elsinga

KWR Water Research Institute ( email )

Nieuwegein
Netherlands

Alexander Graf

Ludwig Maximilian University of Munich (LMU) ( email )

Marchionini Street 15
Munich, 81377
Germany

Leo Heijnen

KWR Water Research Institute ( email )

Johannes Ho

TZW: DVGW-Technologiezentrum Wasser ( email )

Mariana Kluge

Technische Universität München (TUM) ( email )

Stefan Krebs

Ludwig-Maximilians-Universität München, LAFUGA Genomics, Gene Center ( email )

Geschwister-Scholl-Platz 1
Munich, DE Bavaria 80539
Germany

Claudia Stange

TZW: DVGW-Technologiezentrum Wasser ( email )

Anna Uchaikina

Technische Universität München (TUM) ( email )

Regina Dolny

RWTH Aachen University ( email )

Templergraben 55
52056 Aachen, 52056
Germany

Christian Wurzbacher

Technische Universität München (TUM) ( email )

Jörg E. Drewes

Technische Universität München (TUM) ( email )

Gertjan Medema

KWR Water Research Institute ( email )

Nieuwegein
Netherlands

Andreas Tiehm

TZW: DVGW-Technologiezentrum Wasser ( email )

Sandra Ciesek

Goethe University Frankfurt - Institute for Medical Virology ( email )

Paul-Ehrlich-Str. 40
Frankfurt, 60596
Germany

Burkhard Teichgräber

Emschergenossenschaft/Lippeverban ( email )

Thomas Wintgens

RWTH Aachen University ( email )

Templergraben 55
52056 Aachen, 52056
Germany

Frank-Andreas Weber

FiW e.V., Aachen, Germany ( email )

Marek Widera (Contact Author)

Goethe University Frankfurt ( email )

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