Interlaboratory comparison using inactivated SARS-CoV-2 variants as a feasible tool for quality control in COVID-19 wastewater monitoring
The Science of The Total Environment, vol. 903, pp. 166540
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. All PCR-laboratories reported SARS-CoV-2 genome copy equivalents (GCE) for all spiked samples with a mean intra- and inter-laboratory variability of 19 % and 104 %, respectively, largely reproducing the spike-in scheme. 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 highly heterogeneous isolation and analysis procedures, overall SARS-CoV-2 GCE and mutations were determined accurately. 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.
Authors 23
-
Goethe University Frankfurt · University Hospital Frankfurt
Affiliation as printed
Goethe University Frankfurt, University Hospital, Institute for Medical Virology, Paul-Ehrlich-Str. 40, D-60596 Frankfurt, Germany
-
Affiliation as printed
Emschergenossenschaft/Lippeverband, Kronprinzenstraße 24, D-45128 Essen, Germany
-
Affiliation as printed
Ruhrverband, Kronprinzenstraße 37, D-45128 Essen, Germany
-
Affiliation as printed
FiW e.V., Research Institute for Water Management and Climate Future at RWTH Aachen University, Kackertstraße 15-17, D-52056 Aachen, Germany
-
Ludwig-Maximilians-Universität München
Affiliation as printed
Laboratory for Functional Genome Analysis, Gene Center, LMU München, Feodor-Lynen-Straße 25, D-81377 Munich, Germany
-
Affiliation as printed
KWR Water Research Institute, Groningenhaven 7, 3433 PE Nieuwegein, the Netherlands
-
Ludwig-Maximilians-Universität München
Affiliation as printed
Laboratory for Functional Genome Analysis, Gene Center, LMU München, Feodor-Lynen-Straße 25, D-81377 Munich, Germany
-
Affiliation as printed
KWR Water Research Institute, Groningenhaven 7, 3433 PE Nieuwegein, the Netherlands
-
DVGW - Technologiezentrum Wasser
Affiliation as printed
TZW: DVGW-Technologiezentrum Wasser, Karlsruher Str. 84, 76139 Karlsruhe, Germany
-
Technical University of Munich
Affiliation as printed
Chair of Urban Water Systems Engineering, Technical University of Munich, Am Coulombwall 3, D-85748 Garching, Germany
-
Ludwig-Maximilians-Universität München
Affiliation as printed
Laboratory for Functional Genome Analysis, Gene Center, LMU München, Feodor-Lynen-Straße 25, D-81377 Munich, Germany
-
DVGW - Technologiezentrum Wasser
Affiliation as printed
TZW: DVGW-Technologiezentrum Wasser, Karlsruher Str. 84, 76139 Karlsruhe, Germany
-
Technical University of Munich
Affiliation as printed
Chair of Urban Water Systems Engineering, Technical University of Munich, Am Coulombwall 3, D-85748 Garching, Germany
-
Affiliation as printed
Institute of Environmental Engineering, RWTH Aachen University, Mies-van-der-Rohe-Strasse 1, D-52074 Aachen, Germany
-
Technical University of Munich
Affiliation as printed
Chair of Urban Water Systems Engineering, Technical University of Munich, Am Coulombwall 3, D-85748 Garching, Germany
-
Technical University of Munich
Affiliation as printed
Chair of Urban Water Systems Engineering, Technical University of Munich, Am Coulombwall 3, D-85748 Garching, Germany
-
Affiliation as printed
KWR Water Research Institute, Groningenhaven 7, 3433 PE Nieuwegein, the Netherlands
-
DVGW - Technologiezentrum Wasser
Affiliation as printed
TZW: DVGW-Technologiezentrum Wasser, Karlsruher Str. 84, 76139 Karlsruhe, Germany
-
Goethe University Frankfurt · Fraunhofer Institute for Translational Medicine and Pharmacology · German Center for Infection Research · University Hospital Frankfurt
Affiliation as printed
Goethe University Frankfurt, University Hospital, Institute for Medical Virology, Paul-Ehrlich-Str. 40, D-60596 Frankfurt, Germany; German Center for Infection Research (DZIF), 38124 Braunschweig, Germany; Fraunhofer Institute for Translational Medicine and Pharmacology ITMP, Theodor-Stern-Kai 7, D 60595 Frankfurt am Main, Germany
Fraunhofer Institute for Translational Medicine and Pharmacology ITMP, Theodor-Stern-Kai 7, D 60595 Frankfurt am Main, Germany
German Center for Infection Research (DZIF), 38124 Braunschweig, Germany
Goethe University Frankfurt, University Hospital, Institute for Medical Virology, Paul-Ehrlich-Str. 40, D-60596 Frankfurt, Germany
-
Affiliation as printed
Emschergenossenschaft/Lippeverband, Kronprinzenstraße 24, D-45128 Essen, Germany
-
Affiliation as printed
Institute of Environmental Engineering, RWTH Aachen University, Mies-van-der-Rohe-Strasse 1, D-52074 Aachen, Germany
-
Affiliation as printed
FiW e.V., Research Institute for Water Management and Climate Future at RWTH Aachen University, Kackertstraße 15-17, D-52056 Aachen, Germany
-
Goethe University Frankfurt · University Hospital Frankfurt
Affiliation as printed
Goethe University Frankfurt, University Hospital, Institute for Medical Virology, Paul-Ehrlich-Str. 40, D-60596 Frankfurt, Germany. Electronic address: marek.widera@kgu.de
Cited by 11 stored of 11
11 results
No patents citing this paper on Lens.org (checked 2026-10-06).