Comparison of virus concentration methods for the RT-qPCR-based recovery of murine hepatitis virus, a surrogate for SARS-CoV-2 from untreated wastewater

Ahmed, Warish; Bertsch, Paul M.; Bivins, Aaron; Bibby, Kyle; Farkas, Kata; Gathercole, Amy; Haramoto, Eiji; Gyawali, Pradip; Korajkic, Asja; McMinn, Brian R.; Mueller, Jochen F.; Simpson, Stuart L.; Smith, Wendy; Symonds, Erin M.; Thomas, Kevin V.; Verhagen, Rory; Kitajima, Masaaki

doi:10.1016/j.scitotenv.2020.139960

Cited by 457 publications

(549 citation statements)

References 38 publications

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“…However, a complete quantification of viral titers needs further investigation of recovery rates for each step including the pasteurization, filtering, viral precipitation, RNA extraction, and reverse transcription. Recently, Ahmed et al used the beta-coronavirus murine hepatitis virus (MHV) as a closely related surrogate for SARS-CoV-2 to quantify the recovery rates from wastewater; using various methods including PEG precipitation, they found that MHV recoveries ranged from 26.7 to 65.7% ( 20 ). To adjust the variations from the initial sample concentrations and RNA losses during these experimental procedures, we used PMMoV, the most abundant RNA virus in human stool ( 13 , 16 ), as a reference.…”

Section: Discussionmentioning

confidence: 99%

SARS-CoV-2 Titers in Wastewater Are Higher than Expected from Clinically Confirmed Cases

et al. 2020

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Wastewater surveillance represents a complementary approach to clinical surveillance to measure the presence and prevalence of emerging infectious diseases like the novel coronavirus SARS-CoV-2. This innovative data source can improve the precision of epidemiological modeling to understand the penetrance of SARS-CoV-2 in specific vulnerable communities. Here, we tested wastewater collected at a major urban treatment facility in Massachusetts and detected SARS-CoV-2 RNA from the N gene at significant titers (57 to 303 copies per ml of sewage) in the period from 18 to 25 March 2020 using RT-qPCR. We validated detection of SARS-CoV-2 by Sanger sequencing the PCR product from the S gene. Viral titers observed were significantly higher than expected based on clinically confirmed cases in Massachusetts as of 25 March. Our approach is scalable and may be useful in modeling the SARS-CoV-2 pandemic and future outbreaks. IMPORTANCE Wastewater-based surveillance is a promising approach for proactive outbreak monitoring. SARS-CoV-2 is shed in stool early in the clinical course and infects a large asymptomatic population, making it an ideal target for wastewater-based monitoring. In this study, we develop a laboratory protocol to quantify viral titers in raw sewage via qPCR analysis and validate results with sequencing analysis. Our results suggest that the number of positive cases estimated from wastewater viral titers is orders of magnitude greater than the number of confirmed clinical cases and therefore may significantly impact efforts to understand the case fatality rate and progression of disease. These data may help inform decisions surrounding the advancement or scale-back of social distancing and quarantine efforts based on dynamic wastewater catchment-level estimations of prevalence.

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Section: Discussionmentioning

confidence: 99%

SARS-CoV-2 Titers in Wastewater Are Higher than Expected from Clinically Confirmed Cases

et al. 2020

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“…For example, Poliovirus, Norovirus and Hepatitis A are non-enveloped viruses, while SARS-CoV-2 is enveloped, which may influence its ability to be recovered by established methods. Hence, it is advisable to compare different methods using surrogates like mouse hepatitis virus, a group 2 coronavirus, or actual samples ( Ahmed et al, 2020b ).…”

Section: The Case For Sustained Observations Of Wastewater Systemsmentioning

confidence: 99%

Making waves: Wastewater surveillance of SARS-CoV-2 for population-based health management

et al. 2020

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Worldwide, clinical data remain the gold standard for disease surveillance and tracking. However, such data are limited due to factors such as reporting bias and inability to track asymptomatic disease carriers. Disease agents are excreted in the urine and feces of infected individuals regardless of disease symptom severity. Wastewater surveillance-that is, monitoring disease via human effluent-represents a valuable complement to clinical approaches. Because wastewater is relatively inexpensive and easy to collect and can be monitored at different levels of population aggregation as needed, wastewater surveillance can offer a real-time, cost-effective view of a community's health that is independent of biases associated with case-reporting. For SARS-CoV-2 and other disease-causing agents we envision an aggregate wastewatermonitoring system at the level of a wastewater treatment plant and exploratory or confirmatory monitoring of the sewerage system at the neighborhood scale to identify or confirm clusters of infection or assess impact of control measures where transmission has been established. Implementation will require constructing a framework with collaborating government agencies, public or private utilities, and civil society organizations for appropriate use of data collected from wastewater, identification of an appropriate scale of sample collection and aggregation to balance privacy concerns and risk of stigmatization with public health preservation, and consideration of the social implications of wastewater surveillance.

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“…More recently, WBE has been proposed for the early detection of viral outbreaks [ [74] , [75] , [76] , [77] , [78] , [79] , [80] , [81] , [82] ] as well as for the persistence of viral indicators in the aquatic environment [ 75 , 83 ]. The methods used are high throughput RNA sequencing (RNA-Seq) more commonly than PCR (qRT-PCR) [ 75 , 84 ]. In 2019, O’Brien and Xagoraraki [ 74 , 85 ] underlined the importance of wastewater data collection to understand spatial and temporal evolution of viral diseases.…”

Section: Wastewater Based Epidemiologymentioning

confidence: 99%

The role of wastewater treatment plants as tools for SARS-CoV-2 early detection and removal

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Jasim

Johnson

et al. 2020

Journal of Water Process Engineering

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Comparison of virus concentration methods for the RT-qPCR-based recovery of murine hepatitis virus, a surrogate for SARS-CoV-2 from untreated wastewater

Cited by 457 publications

References 38 publications

SARS-CoV-2 Titers in Wastewater Are Higher than Expected from Clinically Confirmed Cases

SARS-CoV-2 Titers in Wastewater Are Higher than Expected from Clinically Confirmed Cases

Making waves: Wastewater surveillance of SARS-CoV-2 for population-based health management

The role of wastewater treatment plants as tools for SARS-CoV-2 early detection and removal

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