Respiratory Syncytial Virus (RSV) RNA in Wastewater Settled Solids Reflects RSV Clinical Positivity Rates

Hughes, Bridgette; Duong, Dorothea; White, Bradley J.; Wigginton, Krista; Chan, Elana M. G.; Wolfe, Marlene K.; Boehm, Alexandria B.

doi:10.1021/acs.estlett.1c00963

Cited by 93 publications

(88 citation statements)

References 31 publications

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“…Future experiments should investigate the decay of other targets used for wastewater-based epidemiology including respiratory syncytial virus (RSV). 36 …”

Section: Discussionmentioning

confidence: 99%

Persistence of Endogenous SARS-CoV-2 and Pepper Mild Mottle Virus RNA in Wastewater-Settled Solids

Roldan-Hernandez

Duong²,

Boehm

2022

ACS EST Water

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Limited information is available on the decay rate of endogenous SARS-CoV-2 and pepper mild mottle virus (PMMoV) RNA in wastewater and primary settled solids, potentially limiting an understanding of how transit or holding times within wastewater infrastructure might impact RNA measurements and their relationship to community COVID-19 infections. In this study, primary settled solids samples were collected from two wastewater treatment plants in the San Francisco Bay Area. Samples were thoroughly mixed, aliquoted into subsamples, and stored at 4, 22, and 37 °C for 10 days. The concentrations of SARS-CoV-2 (N1 and N2 targets) and PMMoV RNA were measured using an RT-ddPCR. Limited decay (<1 log 10 reduction) was observed in the detection of viral RNA targets at all temperature conditions, suggesting that SARS-CoV-2 and PMMoV RNA can be highly persistent in solids. First-order decay rate constants ranged from 0.011 to 0.098 day –1 for SARS-CoV-2 RNA and from 0.010 to 0.091 day –1 for PMMoV RNA depending on the temperature conditions. A slower decay was observed for SARS-CoV-2 RNA in primary settled solids compared to previously reported decay in wastewater influent. Further research is needed to understand if solid content and wastewater characteristics might influence the persistence of viral RNA targets.

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“…Future experiments should investigate the decay of other targets used for wastewater-based epidemiology including respiratory syncytial virus (RSV). 36 …”

Section: Discussionmentioning

confidence: 99%

Persistence of Endogenous SARS-CoV-2 and Pepper Mild Mottle Virus RNA in Wastewater-Settled Solids

Roldan-Hernandez

Duong²,

Boehm

2022

ACS EST Water

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“…Wastewater has also been long established as a public health tool to monitor for poliovirus circulation, and monitoring of SARS-CoV-2, the causative virus of Coronavirus disease 2019 (COVID-19), in wastewater has been reliably used by public health throughout the COVID-19 pandemic. Concentrations of SARS-CoV-2 viral RNA are strongly correlated with case incidence [5][6][7] , and recent studies show this is also the case for respiratory viruses such as Influenza A and RSV 8,9 . Evidence for these use cases have been established over the years, improving interpretation of results and increasing confidence for use in public health.…”

Section: Introductionmentioning

confidence: 87%

Detection of monkeypox viral DNA in a routine wastewater monitoring program

Wolfe

Duong

Hughes

et al. 2022

Preprint

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Wastewater represents a composite biological sample from the entire contributing population. People infected with monkeypox excrete monkeypox virus DNA via skin lesions, saliva, feces and urine and these can enter the wastewater via toilets, sinks, and shower drains. To test whether monkeypox can be detected and monitored in wastewater during a period when publicly reported monkey cases in the region were increasing, we deployed digital PCR assays that target genomic DNA from the monkeypox virus in our routine, ongoing wastewater surveillance program in the Greater Bay Area of California, USA. We measured monkeypox virus DNA daily in settled solids samples from nine wastewater plants over the period of approximately 4 weeks. During that period, we detected monkeypox virus DNA in wastewater solids at nearly all the wastewater plants we routinely sample. Frequency of occurrence and concentrations were highest at plants serving San Francisco County. To confirm the presence of monkeypox DNA, we used two assays that target distinct sequences on the monkeypox genome on a subset of samples and results from both assays were in close agreement strongly suggesting true positives in the wastewater. Additionally, we show that concentrations of monkeypox DNA is 103 times higher in the solid fraction compared to the liquid fraction of wastewater on a mass-equivalent basis.

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“… 4 , 5 As an alternative, wastewater-based surveillance (WBS) is an efficient approach with great potential for warning of infectious disease outbreaks and transmission. 6 , 7 Accordingly, in March 2020, we also proposed that wastewater analysis can be used for the surveillance of the COVID-19 pandemic. 8 Subsequently, SARS-CoV-2 has been successfully detected in wastewater in many countries, including America, India, China, Canada, and some European countries.…”

Section: Introductionmentioning

confidence: 99%

Paper Device Combining CRISPR/Cas12a and Reverse-Transcription Loop-Mediated Isothermal Amplification for SARS-CoV-2 Detection in Wastewater

Cao

Mao

Fang

et al. 2022

Environ. Sci. Technol.

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Wastewater-based surveillance of the COVID-19 pandemic holds great promise; however, a point-of-use detection method for SARS-CoV-2 in wastewater is lacking. Here, a portable paper device based on CRISPR/Cas12a and reverse-transcription loop-mediated isothermal amplification (RT-LAMP) with excellent sensitivity and specificity was developed for SARS-CoV-2 detection in wastewater. Three primer sets of RT-LAMP and guide RNAs (gRNAs) that could lead Cas12a to recognize target genes via base pairing were used to perform the high-fidelity RT-LAMP to detect the N, E, and S genes of SARS-CoV-2. Due to the trans-cleavage activity of CRISPR/Cas12a after high-fidelity amplicon recognition, carboxyfluorescein-ssDNA-Black Hole Quencher-1 and carboxyfluorescein-ssDNA-biotin probes were adopted to realize different visualization pathways via a fluorescence or lateral flow analysis, respectively. The reactions were integrated into a paper device for simultaneously detecting the N, E, and S genes with limits of detection (LODs) of 25, 310, and 10 copies/mL, respectively. The device achieved a semiquantitative analysis from 0 to 310 copies/mL due to the different LODs of the three genes. Blind experiments demonstrated that the device was suitable for wastewater analysis with 97.7% sensitivity and 82% semiquantitative accuracy. This is the first semiquantitative endpoint detection of SARS-CoV-2 in wastewater via different LODs, demonstrating a promising point-of-use method for wastewater-based surveillance.

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Respiratory Syncytial Virus (RSV) RNA in Wastewater Settled Solids Reflects RSV Clinical Positivity Rates

Cited by 93 publications

References 31 publications

Persistence of Endogenous SARS-CoV-2 and Pepper Mild Mottle Virus RNA in Wastewater-Settled Solids

Persistence of Endogenous SARS-CoV-2 and Pepper Mild Mottle Virus RNA in Wastewater-Settled Solids

Detection of monkeypox viral DNA in a routine wastewater monitoring program

Paper Device Combining CRISPR/Cas12a and Reverse-Transcription Loop-Mediated Isothermal Amplification for SARS-CoV-2 Detection in Wastewater

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