| Autor: |
Wilhelm A; Institute for Medical Virology, University Hospital, Goethe University Frankfurt, Paul-Ehrlich-Str. 40, D-60596 Frankfurt, Germany., Agrawal S; Institute IWAR, Water and Environmental Biotechnology, Technical University of Darmstadt, Franziska-Braun-Str. 7, D-64287 Darmstadt, Germany., Schoth J; Emschergenossenschaft/Lippeverband, Kronprinzenstraße 24, D-45128 Essen, Germany., Meinert-Berning C; Ruhrverband, Kronprinzenstraße 37, D-45128 Essen, Germany., Bastian D; FiW e.V., Research Institute for Water Management and Climate Future at RWTH Aachen University, Kackertstraße 15-17, D-52056 Aachen, Germany., Orschler L; Institute IWAR, Water and Environmental Biotechnology, Technical University of Darmstadt, Franziska-Braun-Str. 7, D-64287 Darmstadt, Germany., Ciesek S; Institute for Medical Virology, University Hospital, Goethe University Frankfurt, Paul-Ehrlich-Str. 40, D-60596 Frankfurt, Germany.; German Center for Infection Research (DZIF), D-38124 Braunschweig, Germany.; Fraunhofer Institute for Translational Medicine and Pharmacology ITMP, Theodor Stern Kai 7, D-60595 Frankfurt am Main, Germany., Teichgräber B; Emschergenossenschaft/Lippeverband, Kronprinzenstraße 24, D-45128 Essen, Germany., Wintgens T; FiW e.V., Research Institute for Water Management and Climate Future at RWTH Aachen University, Kackertstraße 15-17, D-52056 Aachen, Germany.; Institute of Environmental Engineering, RWTH Aachen University, Mies-van-der-Rohe-Strasse 1, D-52074 Aachen, Germany., Lackner S; Institute IWAR, Water and Environmental Biotechnology, Technical University of Darmstadt, Franziska-Braun-Str. 7, D-64287 Darmstadt, Germany., Weber FA; FiW e.V., Research Institute for Water Management and Climate Future at RWTH Aachen University, Kackertstraße 15-17, D-52056 Aachen, Germany., Widera M; Institute for Medical Virology, University Hospital, Goethe University Frankfurt, Paul-Ehrlich-Str. 40, D-60596 Frankfurt, Germany. |
| Abstrakt: |
Wastewater-based SARS-CoV-2 epidemiology (WBE) has been established as an important tool to support individual testing strategies. The Omicron sub-variants BA.4/BA.5 have spread globally, displacing the preceding variants. Due to the severe transmissibility and immune escape potential of BA.4/BA.5, early monitoring was required to assess and implement countermeasures in time. In this study, we monitored the prevalence of SARS-CoV-2 BA.4/BA.5 at six municipal wastewater treatment plants (WWTPs) in the Federal State of North Rhine-Westphalia (NRW, Germany) in May and June 2022. Initially, L452R-specific primers/probes originally designed for SARS-CoV-2 Delta detection were validated using inactivated authentic viruses and evaluated for their suitability for detecting BA.4/BA.5. Subsequently, the assay was used for RT-qPCR analysis of RNA purified from wastewater obtained twice a week at six WWTPs. The occurrence of L452R carrying RNA was detected in early May 2022, and the presence of BA.4/BA.5 was confirmed by variant-specific single nucleotide polymorphism PCR (SNP-PCR) targeting E484A/F486V and NGS sequencing. Finally, the mutant fractions were quantitatively monitored by digital PCR, confirming BA.4/BA.5 as the majority variant by 5 June 2022. In conclusion, the successive workflow using RT-qPCR, variant-specific SNP-PCR, and RT-dPCR demonstrates the strength of WBE as a versatile tool to rapidly monitor variants spreading independently of individual test capacities. |
