Validating and optimizing the method for molecular detection and quantification of SARS-CoV-2 in wastewater

Qiu, Yuanyuan; Yu, Jiaao; Lee, Bonita E.; Gao, Tiejun; Ashbolt, Nicholas J.; Hrudey, Steve E.; Diggle, Mathew; Maal‐Bared, Rasha; Pang, Xiaoli

doi:10.1016/j.scitotenv.2021.151434

Cited by 40 publications

(34 citation statements)

References 51 publications

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“…Our laboratory is currently conducting a large-scale wastewater monitoring project for COVID-19 that includes auto sampling at multiple wastewater treatment plants and smaller catchments [ 17 ]. Due to the disadvantages of auto sampling, we are interested in determining whether passive sampling poses a viable option for detecting SARS-CoV-2 in small catchments.…”

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confidence: 99%

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Comparison of Auto Sampling and Passive Sampling Methods for SARS-CoV-2 Detection in Wastewater

Wilson

Qiu

et al. 2022

Pathogens

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Wastewater-based surveillance is emerging as an important tool for the COVID-19 pandemic trending. Current methods of wastewater collection, such as grab and auto-composite sampling, have drawbacks that impede effective surveillance, especially from small catchments with limited accessibility. Passive samplers, which are more cost-effective and require fewer resources to process, are promising candidates for monitoring wastewater for SARS-CoV-2. Here, we compared traditional auto sampling with passive sampling for SARS-CoV-2 detection in wastewater. A torpedo-style 3D-printed passive sampler device containing both cotton swabs and electronegative filter membranes was used. Between April and June 2021, fifteen passive samplers were placed at a local hospital’s wastewater outflow alongside an autosampler. Reverse transcription and quantitative polymerase chain reaction (RT-qPCR) was used to detect SARS-CoV-2 in the samples after processing and RNA extraction. The swab and membrane of the passive sampler showed similar detection rates and cycle threshold (Ct) values for SARS-CoV-2 RNA for the N1 and N2 gene targets. The passive method performed as well as the grab/auto sampling, with no significant differences between N1 and N2 Ct values. There were discrepant results on two days with negative grab/auto samples and positive passive samples, which might be related to the longer duration of passive sampling in the study. Overall, the passive sampler was rapid, reliable, and cost-effective, and could be used as an alternative sampling method for the detection of SARS-CoV-2 in wastewater.

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confidence: 99%

“…The samples were concentrated using Centricon ® Plus-70 centrifugal ultrafilters (30-kDa MWCO, Millipore, Burlington, MA, USA) [ 17 ]. Briefly, the pH of the samples was adjusted to 9.6–10.0, then vortexed for 30 s to release solid-bound virus and RNA into solution, followed by centrifugation at 4500× g for 10 min.…”

mentioning

confidence: 99%

Comparison of Auto Sampling and Passive Sampling Methods for SARS-CoV-2 Detection in Wastewater

Wilson

Qiu

et al. 2022

Pathogens

Self Cite

View full text Add to dashboard Cite

show abstract

“…Our lab is currently conducting a large-scale wastewater monitoring project for COVID-19 that includes autosampling at multiple wastewater treatment plants and smaller catchments [17]. Due to the disadvantages of autosampling, we are interested in determining whether passive sampling poses a viable option for detecting SARS-CoV-2 in small catchments.…”

mentioning

confidence: 99%

“…The samples were concentrated using Centricon® Plus-70 centrifugal ultrafilters (30-kDa MWCO, Millipore) [17]. Briefly, the pH of the samples was adjusted to 9.6-10.0, then vortexed for 30 seconds to release solid-bound virus and RNA into solution, followed by centrifugation at 4500 x g for 10 min.…”

mentioning

confidence: 99%

“…RNA was extracted from 400 μL of the concentrated wastewater and passive samples using the MagMax TM 96 viral isolation kit with the Kingfisher Flex automatic system to get a final elution volume of 100 μL. One-step reverse transcription quantitative PCR (RT-qPCR) targeting N1 and N2 genes of SARS-CoV-2, as well as pepper mild mottle virus (PMMoV), the fecal indicator, were performed as previously described [17,19]. Each sample was tested for N1 and N2 genes in duplicate.…”

mentioning

confidence: 99%

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Comparison of Auto Sampling and Passive Sampling Methods for SARS-CoV-2 Detection in Wastewater

Wilson¹,

Qiu²,

Yu³

et al. 2022

Preprint

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Wastewater-based surveillance is emerging as an important tool for COVID-19 pandemic trending. Current methods of wastewater collection, such as grab and auto-composite sampling, have drawbacks that impede effective surveillance, especially from small catchments with limited accessibility. Passive samplers, which are more cost-effective and require fewer resources to process, are promising candidates for monitoring wastewater for SARS-CoV-2. Here, we compared traditional auto sampling with passive sampling for SARS-CoV-2 detection in wastewater. Torpedo-style 3D printed passive sampler device containing both cotton swabs and electronegative filter membranes was used. Between April and June 2021, fifteen passive samplers were placed at a local hospital wastewater outflow alongside an auto sampler. Reverse transcription and quantitative polymerase chain reaction (RT-qPCR) was used to detect SARS-CoV-2 in the samples after processing and RNA extraction. The swab and membrane of the passive sampler showed similar detection rates and Ct values for SARS-CoV-2 RNA for the N1 and N2 gene targets. The passive method performed as well as the grab/auto sampling, with no significant differences between N1 and N2 Ct values. There were discrepant results on two days with negative grab/auto samples and positive passive samples, which might be related to the longer duration of passive sampling in the study. Overall, the passive sampler was rapid, reliable and cost-effective, and could be used as an alternative sampling method for the detection of SARS-CoV-2 in wastewater.

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SARS-CoV-2 surveillance in medical and industrial wastewater—a global perspective: a narrative review

Yang

Guo

Møhlenberg

et al. 2023

Environ Sci Pollut Res

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The novel coronavirus SARS-CoV-2 has spread at an unprecedented rate since late 2019, leading to the global COVID-19 pandemic. During the pandemic, being able to detect SARS-CoV-2 in human populations with high coverage quickly is a huge challenge. As SARS-CoV-2 is excreted in human excreta and thus exposed to the aqueous environment through sewers, the goal is to develop an ideal, non-invasive, cost-effective epidemiological method for detecting SARS-CoV-2. Wastewater surveillance has gained widespread interest and is increasingly being investigated as an effective early warning tool for monitoring the spread and evolution of the virus. This review emphasizes important findings on SARS-CoV-2 wastewater-based epidemiology (WBE) in different continents and techniques used to detect SARS-CoV-2 in wastewater during the period 2020–2022. The results show that WBE is a valuable population-level method for monitoring SARS-CoV-2 and is a valuable early warning alert. It can assist policymakers in formulating relevant policies to avoid the negative impacts of early or delayed action. Such strategy can also help avoid unnecessary wastage of medical resources, rationalize vaccine distribution, assist early detection, and contain large-scale outbreaks.

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Validating and optimizing the method for molecular detection and quantification of SARS-CoV-2 in wastewater

Cited by 40 publications

References 51 publications

Comparison of Auto Sampling and Passive Sampling Methods for SARS-CoV-2 Detection in Wastewater

Comparison of Auto Sampling and Passive Sampling Methods for SARS-CoV-2 Detection in Wastewater

Comparison of Auto Sampling and Passive Sampling Methods for SARS-CoV-2 Detection in Wastewater

SARS-CoV-2 surveillance in medical and industrial wastewater—a global perspective: a narrative review

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