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Optimization and Application of a Multiplex Digital PCR Assay for the Detection of SARS-CoV-2 Variants of Concern in Belgian Influent Wastewater.

Boogaerts, Tim
Van den Bogaert, Siel
El Masri, Diala
De Roeck, Naomi
Lahousse, Lies
van Nuijs, Alexander L.N.
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Abstract

Since the beginning of the COVID-19 pandemic, the wastewater-based epidemiology (WBE) of SARS-CoV-2 has been used as a complementary indicator to follow up on the trends in the COVID-19 spread in Belgium and in many other countries. To further develop the use of WBE, a multiplex digital polymerase chain reaction (dPCR) assay was optimized, validated and applied for the measurement of emerging SARS-CoV-2 variants of concern (VOC) in influent wastewater (IWW) samples. Key mutations were targeted in the different VOC strains, including SΔ69/70 deletion, N501Y, SΔ241 and SΔ157. The presented bioanalytical method was able to distinguish between SARS-CoV-2 RNA originating from the wild-type and B.1.1.7, B.1.351 and B.1.617.2 variants. The dPCR assay proved to be sensitive enough to detect low concentrations of SARS-CoV-2 RNA in IWW since the limit of detection of the different targets ranged between 0.3 and 2.9 copies/µL. This developed WBE approach was applied to IWW samples originating from different Belgian locations and was able to monitor spatio-temporal changes in the presence of targeted VOC strains in the investigated communities. The present dPCR assay developments were realized to bring added-value to the current national WBE of COVID-19 by also having the spatio-temporal proportions of the VoC in presence in the wastewaters.

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Date
2022-12-31
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Peer reviewed scientific article
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Keywords
Belgium, COVID-19, Humans, Multiplex Polymerase Chain Reaction, Pandemics, RNA, Viral, SARS-CoV-2, Waste water
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Topic(s)
Coronavirus #22355#
Disease caused by SARS-CoV-2 (COVID-19) #22545#
Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) #22356#
Wastewater-based epidemiological surveillance #22654#
Related project
Wastewater #1000706#
WWSURV #1000522#
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