Examining the stability of viral RNA and DNA in wastewater: Effects of storage time, temperature, and freeze-thaw cycles

Water Research, Journal Year: 2024, Volume and Issue: 259, P. 121879 - 121879

Published: June 3, 2024

Wastewater-based epidemiology (WBE) has been demonstrably successful as a relatively unbiased tool for monitoring levels of SARS-CoV-2 virus circulating in communities during the COVID-19 pandemic. Accumulated biobanks wastewater samples allow retrospective exploration spatial and temporal trends public health indicators such chemicals, viruses, antimicrobial resistance genes, possible emergence novel human or zoonotic pathogens. We investigated resilience to time, temperature, freeze-thaw cycles, plus optimal storage conditions maintain stability genetic material (RNA/DNA) viral +ssRNA (Envelope - E, Nucleocapsid N Spike protein S genes SARS-CoV-2), dsRNA (Phi6 phage) circular dsDNA (crAssphage) wastewater. Samples consisted (i) processed extracted samples, (ii) distilled water (iii) raw, unprocessed samples. were stored at -80 °C, -20 4 20 °C 10 days, going through up cycles (once per day). Sample was measured using reverse transcription quantitative PCR, automated electrophoresis, short-read whole genome sequencing. Exploring different areas demonstrated that gene showed greater sensitivity than E genes. Investigating surrogate normalisation viruses Phi6 remains stable comparison laboratory setting crAssphage resilient temperature variation. Recovery raw significantly when which supported by sequencing data all both time negatively impacted metrics. Historical extracts re-quantified 12, 14 16 months after original quantification no major changes. This study highlights importance fast processing extraction following are robust range temperatures.

Language: Английский

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Near-source passive sampling for monitoring viral outbreaks within a university residential setting

Epidemiology and Infection, Journal Year: 2024, Volume and Issue: 152

Published: Jan. 1, 2024

Abstract Wastewater-based epidemiology (WBE) has proven to be a powerful tool for the population-level monitoring of pathogens, particularly severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). For assessment, several wastewater sampling regimes and methods viral concentration have been investigated, mainly targeting SARS-CoV-2. However, use passive samplers in near-source environments range viruses is still under-investigated. To address this, samples were taken at four locations student hall residence. These chosen as an exemplar due their high population density perceived risk disease transmission. Viruses investigated SARS-CoV-2 its variants concern (VOCs), influenza viruses, enteroviruses. Sampling was conducted either morning, where place overnight (17 h) during day, with exposure 7 h. We demonstrated usefulness detection VOCs using quantitative polymerase chain reaction (qPCR) next-generation sequencing (NGS). Furthermore, outbreaks A sporadic enteroviruses (some associated enterovirus D68 coxsackieviruses) identified among resident population, providing evidence near-source, in-sewer health communities.

Language: Английский

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Simplifying SARS-CoV-2 wastewater-based surveillance using an automated FDA EUA assay

Microbiology Spectrum, Journal Year: 2025, Volume and Issue: unknown

Published: Feb. 25, 2025

ABSTRACT Wastewater-based surveillance (WBS) can track the spread of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in communities. Laboratory methods for this testing involve labor-intensive, multi-step processes. This study assessed feasibility performing WBS with an off-label use automated commercial SARS-CoV-2 assay that had received Emergency Use Authorization human diagnostic from United States Food and Drug Administration (FDA EUA). Twenty-four-hour composite samples primary influent wastewater seven municipalities New Hampshire Vermont were collected between September 2020 February 2021, centrifuged upon receipt. An aliquot fresh supernatant was immediately tested Abbott m 2000 RealTi e (Abbott Molecular, Des Plaines, IL, USA). Corresponding aliquots then stored at −80°C until they thawed, polyethylene glycol (PEG) concentrated, by two PCR-based laboratory-developed tests (LDTs). Wastewater (103) successful detection viral RNA all three methods. Bland-Altman analysis showed overall concordant results a bias −0.13 −0.42 log copies/mL detected FDA EUA compared to LDTs. Specimen stability assessment demonstrated decrease 33.9% measurable after freeze-thaw cycles. using on platform performed comparably but more efficient workflow when sample-to-answer method could save time labor testing, further validation its ability quantitate is necessary. IMPORTANCE proof-of-principle evaluates (FDA) (EUA) surveillance. Compared standard, currently offers requiring less faster turnaround time.

Language: Английский

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Evaluation of different molecular systems for detection and quantification of SARS-CoV-2 RNA from wastewater samples

Journal of Virological Methods, Journal Year: 2024, Volume and Issue: 328, P. 114956 - 114956

Published: May 23, 2024

Wastewater-based epidemiology has proved to be a suitable approach for tracking the spread of epidemic agents including SARS-CoV-2 RNA. Different protocols have been developed quantitative detection RNA from wastewater samples, but little is known on their performance. In this study we compared three based Reverse Transcription Real Time-PCR (RT-PCR) and one Droplet Digital PCR (ddPCR) 35 samples. Overall, was detected by at least method in 85.7% while 51.4%, 22.8% 8.6% resulted positive with two, or all four methods, respectively. Protocols commercial RT-PCR assays showed an overall higher sensitivity vs. in-house assay. The use more than system, targeting different genes, could helpful increase sensitivity.

Language: Английский

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Wastewater-based surveillance for Hepatitis A virus, Enterovirus, Poliovirus, and SARS-CoV-2 in São Tomé and Príncipe: A pilot study

The Science of The Total Environment, Journal Year: 2024, Volume and Issue: 955, P. 176923 - 176923

Published: Oct. 18, 2024

Wastewater-based surveillance is a valuable tool for monitoring pathogen transmission in communities, especially regions where formal systems are limited.

Language: Английский

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