Evaluation of SARS-CoV-2 concentrations in wastewater and river water samples

Fonseca, Maísa Santos; Machado, Bruna Aparecida Souza; Rolo, Carolina de Araújo; Hodel, Katharine Valéria Saraiva; Almeida, Edna dos Santos; Andrade, Jailson B. de

doi:10.1016/j.cscee.2022.100214

Cited by 13 publications

(7 citation statements)

References 46 publications

(83 reference statements)

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“…In fact, the predictions of viral transmission dynamics based on such data are resistant to changes in the behavior of the public (e.g., testing practices, healthcare-seeking behavior, etc.) (19). On the other hand, certain limitations like environmental conditions (e.g., actual water temperature, dilution of wastewater due to the increased rainfall, the intermittent presence of chemical compounds that can act as PCR inhibitors, and sampling design) can potentially affect the results of WBE (12).…”

Section: Discussionmentioning

confidence: 99%

“…The majority of protocols for SARS-CoV-2 RNA isolation from wastewater use initial centrifugation for the removal of debris prior to processing ( 7 , 27 , 28 ). This step is important for reducing the turbidity of wastewater via removing larger particles and finer particles, which could inhibit PCR reaction as well as improve virus recovery (mainly for samples nearer the limit of detection) ( 7 , 29 – 31 ).…”

Section: Discussionmentioning

confidence: 99%

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Comparison of the methods for isolation and detection of SARS-CoV-2 RNA in municipal wastewater

Lucansky

Samec

Burjanivova

et al. 2023

Front. Public Health

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IntroductionCoronavirus SARS-CoV-2 is a causative agent responsible for the current global pandemic situation known as COVID-19. Clinical manifestations of COVID-19 include a wide range of symptoms from mild (i.e., cough, fever, dyspnea) to severe pneumonia-like respiratory symptoms. SARS-CoV-2 has been demonstrated to be detectable in the stool of COVID-19 patients. Waste-based epidemiology (WBE) has been shown as a promising approach for early detection and monitoring of SARS-CoV-2 in the local population performed via collection, isolation, and detection of viral pathogens from environmental sources.MethodsIn order to select the optimal protocol for monitoring the COVID-19 epidemiological situation in region Turiec, Slovakia, we (1) compared methods for SARS-CoV-2 separation and isolation, including virus precipitation by polyethylene glycol (PEG), virus purification via ultrafiltration (Vivaspin®) and subsequent isolation by NucleoSpin RNA Virus kit (Macherey-Nagel), and direct isolation from wastewater (Zymo Environ Water RNA Kit); (2) evaluated the impact of water freezing on SARS- CoV-2 separation, isolation, and detection; (3) evaluated the role of wastewater filtration on virus stability; and (4) determined appropriate methods including reverse transcription-droplet digital PCR (RT-ddPCR) and real-time quantitative polymerase chain reaction (RT-qPCR) (targeting the same genes, i.e., RdRp and gene E) for quantitative detection of SARS-CoV-2 in wastewater samples.Results(1) Usage of Zymo Environ Water RNA Kit provided superior quality of isolated RNA in comparison with both ultracentrifugation and PEG precipitation. (2) Freezing of wastewater samples significantly reduces the RNA yield. (3) Filtering is counterproductive when Zymo Environ Water RNA Kit is used. (4) According to the specificity and sensitivity, the RT-ddPCR outperforms RT-qPCR.DiscussionThe results of our study suggest that WBE is a valuable early warning alert and represents a non-invasive approach to monitor viral pathogens, thus protects public health on a regional and national level. In addition, we have shown that the sensitivity of testing the samples with a nearer detection limit can be improved by selecting the appropriate combination of enrichment, isolation, and detection methods.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Comparison of the methods for isolation and detection of SARS-CoV-2 RNA in municipal wastewater

Lucansky

Samec

Burjanivova

et al. 2023

Front. Public Health

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“…Previously, our group evaluated and validated the e ciency of four methods to concentrate and determine SARS-CoV-2 from wastewater and wastewater-enriched river water samples. We observed that ultra ltration and PEG 8000 precipitation showed the best SARS-CoV-2 concentration e ciency to wastewater and river water samples, either in aqueous and suspended solids samples and without pretreatment needed to suspended solid removal 30 . Herein we build on our previous work by applied the wastewater surveillance approach, employing PEG 8000 precipitation methodology, to monitor the prevalence of SARS-CoV-2 in raw and treated wastewater samples from 22 municipal wastewater treatment plants (WWTPs) located in Salvador, Bahia State, Brazil.…”

Section: Introductionmentioning

confidence: 87%

“…The recovery of the genetic material of SARS-CoV-2 by concentration using PEG 8000 (which is based on the precipitation analyzes) has shown a better yield when compared to other techniques, such as occulation or ultracentrifugation 30,43,44 . Therefore, a lower concentration of organic matter (or reaction inhibitors) in the treated samples, as well as the application of a concentration technique capable of increasing the e ciencies of viral recovery may explain the superior performance of the positivity rate of the treated samples that were concentrated by PEG 8000 compared to the others analyzed in this study.…”

Section: Successful Detection Of Sars-cov-2 In Wastewater Samplesmentioning

confidence: 99%

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Long-term monitoring of COVID 19 prevalence in raw and treated wastewater in Salvador, a largest capital of Brazilian Northeast

Rolo

Machado

Carmo

et al. 2023

Preprint

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Wastewater-based epidemiology (WBE) becomes an interesting epidemiological approach to monitoring the prevalence of SARS-CoV-2 broadly and non-invasively. Herein, we employing for the first time WBE, associated or not with the PEG 8000 precipitation method, for detection of SARS-CoV-2 in samples of raw or treated wastewater from 22 municipal wastewater treatment stations (WWTPs) located in Salvador, the fourth most populous city in Brazil. Our results demonstrate the success of the application of WBE for detecting SARS-CoV-2 in both types of evaluated samples, regardless the usage of PEG 8000 concentration procedure. Further, an increase in SARS-CoV-2 positivity rate was observed in samples collected in months that presented the highest number of confirmed COVID-19 cases (May/2021, June/2021 and January/2022). While PEG 8000 concentration step was found to significantly increase the positivity rate in treated wastewater samples (p < 0.005), a strong positive correlation (r: 0.84; p < 0.002) between non-concentrated raw wastewater samples with the number of new cases of COVID-19 (April/2021 – February/2022) was observed. In general, the present results reinforce the efficiency of WBE approach to monitor the presence of SARS-CoV-2 in either low- and high-capacity WWTPs. The successful usage of WBE even in raw wastewater samples makes it an interesting low-cost tool for epidemiological surveillance.

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Evaluation of viral concentration and extraction methods for SARS-CoV-2 recovery from wastewater using droplet digital and quantitative RT-PCR

Dimitrakopoulos¹,

Kontou²,

Strati³

et al. 2022

Case Studies in Chemical and Environmental Engineering

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Evaluation of SARS-CoV-2 concentrations in wastewater and river water samples

Cited by 13 publications

References 46 publications

Comparison of the methods for isolation and detection of SARS-CoV-2 RNA in municipal wastewater

Comparison of the methods for isolation and detection of SARS-CoV-2 RNA in municipal wastewater

Long-term monitoring of COVID 19 prevalence in raw and treated wastewater in Salvador, a largest capital of Brazilian Northeast

Evaluation of viral concentration and extraction methods for SARS-CoV-2 recovery from wastewater using droplet digital and quantitative RT-PCR

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