Sewage surveillance for SARS-CoV-2: Molecular detection, quantification, and normalization factors

Mazumder, Payal; Dash, Siddhant; Honda, Ryo; Sonne, Christian; Kumar, Manish

doi:10.1016/j.coesh.2022.100363

Cited by 23 publications

(13 citation statements)

References 67 publications

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“…Hence, WBE offers the possibility of an early warning system for the COVID-19 in the population. Therefore, WBE has rightfully became one of the most potent means for health authorities worldwide to monitor COVID-19 ( 18 ). 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.)…”

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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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%

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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“…Here, we used the microbial FIT framework to depict, for the first time, the loss of spatially distributed information about fecal contributions to the sewer system over travel times using crAssphage genetic markers. Previous WWS research has used crAssphage and other markers (e.g., PMMoV) to normalize SARS-CoV-2 concentrations to account for variability in wastewater responses. − ,, However, our work reveals that information degradation and sewershed characteristics influence crAssphage load, impacting the proportional pathogen prevalence modeled using fecal marker normalization methods. , …”

Section: Resultsmentioning

confidence: 82%

“…Previous WWS research has used crAssphage and other markers (e.g., PMMoV) to normalize SARS-CoV-2 concentrations to account for variability in wastewater responses. [21][22][23][24]54,55 However, our work reveals that information degradation and sewershed characteristics influence crAssphage load, impacting the proportional pathogen prevalence modeled using fecal marker normalization methods. 2,56 We note some advantages of our semimechanistic, semistatistical approach over fully mechanistic models.…”

Section: Implications Limitations and Future Researchmentioning

confidence: 99%

Characterizing Spatial Information Loss for Wastewater Surveillance Using crAssphage: Effect of Decay, Temperature, and Population Mobility

Wiesner-Friedman,

Brinkman,

Wheaton

et al. 2023

Environ. Sci. Technol.

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“…Household and community sewer system pipes deliver the viral gene fragments to wastewater treatment plants (WWTPs), where they accumulate in the raw sewage. Raw sewage, collected from WWTPs, therefore, is a great source of viral gene fragments [23][24][25]28]. The analysis of raw sewage for viral gene fragments has become a popular method for detecting viral spread within a community [29][30][31].…”

Section: Introductionmentioning

confidence: 99%

“…Subsequently, the changes in the microbial composition of wastewater could be tracked for additional biomarkers to strengthen current WBE approach with SARS-CoV-2. Overall, WBE has become an effective and convenient tool used by many local health authorities around the globe [28,[39][40][41].…”

Section: Introductionmentioning

confidence: 99%

SARS-CoV-2 Monitoring in Wastewater Reveals Novel Variants and Biomarkers of Infection

McGowan

Borucki

Omairi

et al. 2022

Viruses

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Wastewater-based epidemiology (WBE) is a popular tool for the early indication of community spread of infectious diseases. WBE emerged as an effective tool during the COVID-19 pandemic and has provided meaningful information to minimize the spread of infection. Here, we present a combination of analyses using the correlation of viral gene copies with clinical cases, sequencing of wastewater-derived RNA for the viral mutants, and correlative analyses of the viral gene copies with the bacterial biomarkers. Our study provides a unique platform for potentially using the WBE-derived results to predict the spread of COVID-19 and the emergence of new variants of concern. Further, we observed a strong correlation between the presence of SARS-CoV-2 and changes in the microbial community of wastewater, particularly the significant changes in bacterial genera belonging to the families of Lachnospiraceae and Actinomycetaceae. Our study shows that microbial biomarkers could be utilized as prediction tools for future infectious disease surveillance and outbreak responses. Overall, our comprehensive analyses of viral spread, variants, and novel bacterial biomarkers will add significantly to the growing body of literature on WBE and COVID-19.

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Sewage surveillance for SARS-CoV-2: Molecular detection, quantification, and normalization factors

Cited by 23 publications

References 67 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

Characterizing Spatial Information Loss for Wastewater Surveillance Using crAssphage: Effect of Decay, Temperature, and Population Mobility

SARS-CoV-2 Monitoring in Wastewater Reveals Novel Variants and Biomarkers of Infection

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