Wilhelm, Alexander ; Agrawal, Shelesh ; Schoth, Jens ; Meinert-Berning, Christina ; Bastian, Daniel ; Orschler, Laura ; Ciesek, Sandra ; Teichgräber, Burkhard ; Wintgens, Thomas ; Lackner, Susanne ; Weber, Frank-Andreas ; Widera, Marek (2022)
Early Detection of SARS-CoV-2 Omicron BA.4 and BA.5 in German Wastewater.
In: Viruses, 2022, 14 (9)
doi: 10.26083/tuprints-00022326
Artikel, Zweitveröffentlichung, Verlagsversion
Kurzbeschreibung (Abstract)
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.
| Typ des Eintrags: | Artikel |
|---|---|
| Erschienen: | 2022 |
| Autor(en): | Wilhelm, Alexander ; Agrawal, Shelesh ; Schoth, Jens ; Meinert-Berning, Christina ; Bastian, Daniel ; Orschler, Laura ; Ciesek, Sandra ; Teichgräber, Burkhard ; Wintgens, Thomas ; Lackner, Susanne ; Weber, Frank-Andreas ; Widera, Marek |
| Art des Eintrags: | Zweitveröffentlichung |
| Titel: | Early Detection of SARS-CoV-2 Omicron BA.4 and BA.5 in German Wastewater |
| Sprache: | Englisch |
| Publikationsjahr: | 2022 |
| Ort: | Darmstadt |
| Publikationsdatum der Erstveröffentlichung: | 2022 |
| Verlag: | MDPI |
| Titel der Zeitschrift, Zeitung oder Schriftenreihe: | Viruses |
| Jahrgang/Volume einer Zeitschrift: | 14 |
| (Heft-)Nummer: | 9 |
| Kollation: | 15 Seiten |
| DOI: | 10.26083/tuprints-00022326 |
| URL / URN: | https://tuprints.ulb.tu-darmstadt.de/22326 |
| Zugehörige Links: | |
| Herkunft: | Zweitveröffentlichung DeepGreen |
| Kurzbeschreibung (Abstract): | 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. |
| Freie Schlagworte: | COVID-19 surveillance, SARS-CoV-2 monitoring, wastewater-based epidemiology (WBE), Omicron, BA.4, BA.5, variant of concern |
| Status: | Verlagsversion |
| URN: | urn:nbn:de:tuda-tuprints-223265 |
| Zusätzliche Informationen: | This article belongs to the Special Issue Molecular Epidemiology of SARS-CoV-2 |
| Sachgruppe der Dewey Dezimalklassifikatin (DDC): | 600 Technik, Medizin, angewandte Wissenschaften > 610 Medizin, Gesundheit 600 Technik, Medizin, angewandte Wissenschaften > 620 Ingenieurwissenschaften und Maschinenbau |
| Fachbereich(e)/-gebiet(e): | 13 Fachbereich Bau- und Umweltingenieurwissenschaften 13 Fachbereich Bau- und Umweltingenieurwissenschaften > Institut IWAR - Wasser- und Abfalltechnik, Umwelt- und Raumplanung 13 Fachbereich Bau- und Umweltingenieurwissenschaften > Institut IWAR - Wasser- und Abfalltechnik, Umwelt- und Raumplanung > Fachgebiet Wasser und Umweltbiotechnologie |
| Hinterlegungsdatum: | 31 Okt 2022 14:29 |
| Letzte Änderung: | 01 Nov 2022 08:44 |
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