TY  - JOUR
A1  - Wilhelm, Alexander
A1  - Agrawal, Shelesh
A1  - Schoth, Jens
A1  - Meinert-Berning, Christina
A1  - Bastian, Daniel
A1  - Orschler, Laura
A1  - Ciesek, Sandra
A1  - Teichgräber, Burkhard
A1  - Wintgens, Thomas
A1  - Lackner, Susanne
A1  - Weber, Frank-Andreas
A1  - Widera, Marek
T1  - Early detection of SARS-CoV-2 Omicron BA.4 and BA.5 in German wastewater
T2  - Viruses
N2  - 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.
KW  - COVID-19 surveillance
KW  - SARS-CoV-2 monitoring
KW  - wastewater-based epidemiology (WBE)
KW  - Omicron
KW  - BA.4
KW  - BA.5
KW  - variant of concern
Y1  - 2022
UR  - http://publikationen.ub.uni-frankfurt.de/frontdoor/index/index/docId/73657
UR  - https://nbn-resolving.org/urn:nbn:de:hebis:30:3-736572
SN  - 1999-4915
VL  - 14
IS  - 9
PB  - MDPI
CY  - Basel
ER  -