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Statistical framework to support the epidemiological interpretation of SARS-CoV-2 concentration in municipal wastewater.

ABSTRACT: The ribonucleic acid (RNA) of the severe acute respiratory syndrome coronavirus 2 (SARS-Cov-2) is detectable in municipal wastewater as infected individuals can shed the virus in their feces. Viral concentration in wastewater can inform the severity of the COVID-19 pandemic but observations can be noisy and sparse and hence hamper the epidemiological interpretation. Motivated by a Canadian nationwide wastewater surveillance data set, unlike previous studies, we propose a novel Bayesian statistical framework based on the theories of functional data analysis to tackle the challenges embedded in the longitudinal wastewater monitoring data. By employing this framework to analyze the large-scale data set from the nationwide wastewater surveillance program covering 15 sampling sites across Canada, we successfully detect the true trends of viral concentration out of noisy and sparsely observed viral concentrations, and accurately forecast the future trajectory of viral concentrations in wastewater. Along with the excellent performance assessment using simulated data, this study shows that the proposed novel framework is a useful statistical tool and has a significant potential in supporting the epidemiological interpretation of noisy viral concentration measurements from wastewater samples in a real-life setting.

SUBMITTER: Dai X

PROVIDER: S-EPMC9355971 | biostudies-literature | 2022 Aug

REPOSITORIES: biostudies-literature

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Statistical framework to support the epidemiological interpretation of SARS-CoV-2 concentration in municipal wastewater.

Dai Xiaotian X Champredon David D Fazil Aamir A Mangat Chand S CS Peterson Shelley W SW Mejia Edgard M EM Lu Xuewen X Chekouo Thierry T

Scientific reports 20220805 1

The ribonucleic acid (RNA) of the severe acute respiratory syndrome coronavirus 2 (SARS-Cov-2) is detectable in municipal wastewater as infected individuals can shed the virus in their feces. Viral concentration in wastewater can inform the severity of the COVID-19 pandemic but observations can be noisy and sparse and hence hamper the epidemiological interpretation. Motivated by a Canadian nationwide wastewater surveillance data set, unlike previous studies, we propose a novel Bayesian statistic ...[more]

PMID: 35931713

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Project description:Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) wastewater-based epidemiology (WBE) has been applied as a complementary approach for spatial tracking of coronavirus disease 2019 cases (COVID-19) as well as early warning of the occurrence of infected populations. The present study presents the result of the monitoring of sanitary sewerage in the municipality of Niterói, a metropolitan region of Rio de Janeiro (Brazil) and its use as a complementary indicator in the surveillance of COVID-19 cases, thus assisting actions of public health from local authorities. Twelve composite raw sewage samples were weekly collected from two wastewater treatment plants (WWTPs) and alternately from 17 sewer pipes (SP) from surrounding neighbourhoods and slums throughout 20 weeks (April 15th to August 25th, 2020). Two hundred twenty-three samples were concentrated using the ultracentrifugation-based method and SARS-CoV-2 RNA detected and quantified by RT-qPCR using primers and probe targeting the N2 genome. SARS-CoV-2 RNA was detected in 84.3% (188/223) of samples with a positive rate ranging from 42% (5/12) in the first week of monitoring to 100% during the peak of epidemic with viral concentration ranging from 3.1 to 7.1 log10 genome copies /100 mL throughout the studied period. Positive rates were higher in WWTPs when compared to SP, being useful tool for monitoring trends in the evolution of the COVID-19 curve, while SP data were more effective when health public interventions were needed. Whole-genome sequencing using Illumina MiSeq System confirmed the lineage of three genomes as B.1.1.33 (clade G) containing the nucleotide substitutions observed in strains that circulate in the Rio de Janeiro during the period of this study. In addition, geoprocessing tool was used to build heat maps based on SARS-CoV-2 data from sewage samples, which were weekly updated and available online to the general population as an indicator of the ongoing epidemic situation in Niterói city, raising public awareness.

Project description:Wastewater1 surveillance of SARS-CoV-2 may be a useful supplement to clinical surveillance as it is shed in feces, there are many asymptomatic cases, and diagnostic testing can have capacity limitations and extended time to results. Although numerous studies have utilized wastewater surveillance for SARS-CoV-2, the methods used were developed and/or standardized for other pathogens. This study evaluates multiple methods for concentration and recovery of SARS-CoV-2 and seeded human coronavirus OC43 from municipal primary wastewater and/or sludge from the Greater Seattle Area (March-July 2020). Methods evaluated include the bag-mediated filtration system (BMFS), with and without Vertrel™ extraction, skimmed milk flocculation, with and without Vertrel™ extraction, polyethylene glycol (PEG) precipitation, ultrafiltration, and sludge extraction. Total RNA was extracted from wastewater concentrates and analyzed for SARS-CoV-2 and OC43 with RT-qPCR. Skimmed milk flocculation without Vertrel™ extraction performed consistently over time and between treatment plants in Seattle-area wastewater with the lowest average OC43 Cq value and smallest variability (24.3; 95% CI: 23.8-24.9), most frequent SARS-CoV-2 detection (48.8% of sampling events), and highest average OC43 percent recovery (9.1%; 95% CI: 6.2-11.9%). Skimmed milk flocculation is also beneficial because it is feasible in low-resource settings. While the BMFS had the highest average volume assayed of 11.9 mL (95% CI: 10.7-13.1 mL), the average OC43 percent recovery was low (0.7%; 95% CI: 0.4-1.0%). Ultrafiltration and PEG precipitation had low average OC43 percent recoveries of 1.0% (95% CI: 0.5-1.6%) and 3.2% (95% CI: 1.3-5.1%), respectively. The slopes and efficiency for the SARS-CoV-2 standard curves were not consistent over time, confirming the need to include a standard curve each run rather than using a single curve for multiple plates. Results suggest that the concentration and detection methods used must be validated for the specific water matrix using a recovery control to assess performance over time.

Project description: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.

Dataset Information

Statistical framework to support the epidemiological interpretation of SARS-CoV-2 concentration in municipal wastewater.

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Statistical framework to support the epidemiological interpretation of SARS-CoV-2 concentration in municipal wastewater.

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