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.
Keywords: Belgium; SARS-CoV-2; digital polymerase chain reaction; variants of concern; wastewater-based epidemiology.
【저자키워드】 SARS-CoV-2, variants of concern, Wastewater-based epidemiology, Belgium, digital polymerase chain reaction, 【초록키워드】 COVID-19, Mutation, Epidemiology, VoC, B.1.351, COVID-19 pandemic, variant, SARS-CoV-2 variant, B.1.617.2, variants, Spread, polymerase chain reaction, B.1.1.7, N501Y, limit of detection, target, SARS-CoV-2 RNA, multiplex, Belgium, Strains, Concentration, strain, complementary, Chain Reaction, wild-type, National, MONITOR, approach, polymerase chain, detect, develop, proportion, investigated, applied, changes in, ranged, 【제목키워드】 detection, wastewater, PCR, Digital, concern, application, optimization, influent,