An alternative approach for bioanalytical assay optimization for wastewater-based epidemiology of SARS-CoV-2

Boogaerts, Tim; Jacobs, Lotte; Roeck, Naomi De; Bogaert, Siel Van den; Aertgeerts, Bert; Lahousse, Lies; Nuijs, Alexander L.N. van; Delputte, Peter

doi:10.1016/j.scitotenv.2021.148043

Cited by 31 publications

(21 citation statements)

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“…Ultracentrifugation and glycine elution was the method of choice to determine the time series of SARS-CoV-2 RNA concentrations as it resulted in significantly higher SARS-CoV-2 N1 concentrations than the Centricon ® Plus-70 ultrafiltration method (p,0.001) (Figure 2). Some authors, when comparing the recovery results of SARS-CoV-2 or enveloped viruses (MHV, BCoV, PMMoV) obtained by different concentration methods, observed that ultrafiltration using Centricon ® Plus-70 had better performance than other methods such as CP-Select™ ultrafiltration (Forés et al 2021), PEG 8000 precipitation (Guerrity et al 2021), Amicon, Macrosep, Vivaspin ultrafiltration, PEG 8000 precipitation (Boogaerts et al 2021), PEG 6000 precipitation (Bertrand et al 2021) and the adsorption-elution method using electronegative membranes (Sherchan et al 2020). Our comparative study was based on a few samples (n¼13) previously filtered on 0.22 μm membranes (not centrifuged) before the Centricon concentration step, a difference that may have reduced the effectiveness of the method but was necessary for the viability test in cell culture.…”

Section: Discussionmentioning

confidence: 99%

One-year surveillance of SARS-CoV-2 in wastewater from vulnerable urban communities in metropolitan São Paulo, Brazil

Barbosa

Garcia

Bruni

et al. 2022

Journal of Water and Health

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The current COVID-19 pandemic has emphasized the vulnerability of communities living in the urban outskirts and informal settlements. The lack of reliable COVID-19 case data highlights the importance and application of wastewater-based epidemiology. This study aimed to monitor the COVID-19 trends in four vulnerable urban communities (slums and low-income neighborhoods) in metropolitan São Paulo by assessing the SARS-CoV-2 RNA viral load in wastewater. We analyzed 160 samples from May 2020 to June 2021 with weekly or fortnightly samplings. The samples were ultracentrifuged with glycine elution and quantified by N1/N2 SARS-CoV-2 RT-qPCR. The results of positivity were 100% (Paraisópolis, Heliópolis and Cidade Tiradentes) and 76.9% (Vila Brasilândia). The new case numbers of COVID-19, counted from the onset of symptoms, positively correlated with SARS-CoV-2 N1 viral loads from the two largest communities (p<0.001). SARS-CoV-2 infectivity was tested in Vero E6 cells after concentration with the two techniques, ultrafiltration (Centricon® Plus-70 10 kDa) and sucrose cushion ultracentrifugation, but none of the evaluated samples presented positive results. Next-generation sequencing (NGS) analysis from samples collected in March and August 2021 revealed the presence of the clade 20 J (lineage P.1) belonging to the most prevalent circulating variant in the country. Our results showed that wastewater surveillance data can be used as complementary indicators to monitor the dynamics and temporal trends of COVID-19. The infectivity test results strengthened the evidence of low risk of infection associated with SARS-CoV-2 in wastewater.

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Section: Discussionmentioning

confidence: 99%

One-year surveillance of SARS-CoV-2 in wastewater from vulnerable urban communities in metropolitan São Paulo, Brazil

Barbosa

Garcia

Bruni

et al. 2022

Journal of Water and Health

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“…In the case of the time-proportional sample collection, a high frequency (less than 10 min) was used to compose the daily IWW samples. Then, 500 mL wastewater was immediately stored after collection at 4 °C to prevent in-sample degradation of SARS-CoV-2 RNA [ 43 , 44 ]. At 24 h upon sample collection, RNA extraction of the IWW samples was performed.…”

Section: Methodsmentioning

confidence: 99%

“…The extraction of SARS-CoV-2 RNA was performed according to a previously validated bioanalytical method [ 43 ]. Briefly, 20 mL of IWW was centrifuged at 4000× g for 30 min to remove solid particles.…”

Section: Methodsmentioning

confidence: 99%

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

Boogaerts

Bogaert

Poelvoorde

et al. 2022

Viruses

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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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“…Storage and transport conditions can affect the detection and quantification of SARS-CoV-2 in wastewater samples because of stability. Various studies provided sufficient evidence that SARS-CoV-2 is persistent in wastewater samples for several days if samples are stored at 4 °C (Ahmed et al 2020b;Boogaerts et al 2021b). As part of the validation process, we have evaluated sample stability.…”

Section: Sample Transport and Storagementioning

confidence: 99%

“…At this moment, there is no ideal external control standard with the same properties of SARS-CoV-2 for quantification available (Boogaerts et al 2021b;Chik et al 2021). Therefore, no spike-in control was in use.…”

Section: Sars-cov-2 Quantificationmentioning

confidence: 99%

Wastewater surveillance of SARS-CoV-2 in Austria: development, implementation, and operation of the Tyrolean wastewater monitoring program

Daleiden

Niederstätter

Steinlechner

et al. 2022

Journal of Water and Health

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Wastewater-based epidemiology (WBE) is an effective approach for tracking information on spatial distribution and temporal trends of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) at the community level. Herein, the development, implementation, and operation of the wastewater monitoring program serving Tyrol – a federal province of Austria – are described. The development of this program was initiated by Tyrolean health authorities at the end of the first phase of the Coronavirus disease 2019 (COVID-19) pandemic (May 2020). In close co-operation with the water sector and academic institutions, efficient and effective workflows and processes for wastewater surveillance were established. The monitoring program went into operation in November 2020. By the end of July 2021, a total of 5,270 wastewater influent samples collected at 43 sites were analyzed. The monitoring program provided valuable insights into the development of the pandemic situation in Tyrol and fulfilled several tasks that are of importance in different phases of the pandemic. It represented an early-warning system, provided independent confirmation of temporal trends in COVID-19 prevalence, enabled the assessment of the effectiveness of measures, alerted about bursts of disease activity, and provided evidence for the absence of COVID-19. These findings underline the importance of establishing national wastewater monitoring programs as a complementary source of information for efficient and effective pandemic management.

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An alternative approach for bioanalytical assay optimization for wastewater-based epidemiology of SARS-CoV-2

Cited by 31 publications

References 50 publications

One-year surveillance of SARS-CoV-2 in wastewater from vulnerable urban communities in metropolitan São Paulo, Brazil

One-year surveillance of SARS-CoV-2 in wastewater from vulnerable urban communities in metropolitan São Paulo, Brazil

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

Wastewater surveillance of SARS-CoV-2 in Austria: development, implementation, and operation of the Tyrolean wastewater monitoring program

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