Novel Workflows for Separate Isolation of Pathogen RNA or DNA from Wastewater: Detection by Innovative and Conventional qPCR

BIO-PROTOCOL, Journal Year: 2025, Volume and Issue: 15(1365)

Published: Jan. 1, 2025

Wastewater-based surveillance (WBS) can provide a wealth of information regarding the health status communities from measurements nucleic acids found in wastewater. Processing workflows for WBS typically include sample collection, primary concentration step, and lysis microbes to release acids, followed by acid purification molecular-based quantification. This manuscript provides beginning end with an emphasis on filtration-based approaches coupled specific extraction processes. Here, two processing are presented, one focusing RNA-specific pathogens other focused DNA-specific within wastewater: 1) The approach, employed analyzing RNA viruses like severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) couples electronegative filtration wastewater placement filter buffer direct extraction. 2) DNA Candida auris, uses size selection membranes during filtration, subsequently buffer, bead-beating, Separate versus isolations have advantage improving detection target pathogen. A novel aspect workflow is lysates, which shows enhanced recoveries, whereas approach requires bead beating prior Novelty also provided new qPCR called Volcano 2nd Generation (V2G), polymerase capable using as template, bypassing reverse transcriptase step normally required qPCR. Key features • Membrane concentrating suspended solids After concentration, optimized separate recovery DNA. Unique utilized perform analysis, foregoing transcription, RNA. Sample products use molecular techniques (e.g., sequencing) generate high-quality, concentrated extracts minimal inhibitors. Validated through COVID-19 where >1,000 samples >3,000 concentrates produced these been created analyzed, published results. complete protocol was used in: J Biomol Tech (2023), DOI: 10.7171/3fc1f5fe.dfa8d906.

Language: Английский

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Evaluation of a field deployable, high-throughput RT-LAMP device as an early warning system for COVID-19 through SARS-CoV-2 measurements in wastewater

The Science of The Total Environment, Journal Year: 2024, Volume and Issue: 944, P. 173744 - 173744

Published: June 5, 2024

Language: Английский

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Academic institution extensive, building-by-building wastewater-based surveillance platform for SARS-CoV-2 monitoring, clinical data correlation, and potential national proxy

Published: Sept. 5, 2024

Abstract Wastewater-based surveillance has been proposed as a cost-effective toolset to generate data about public health status by detecting specific biomarkers in wastewater samples, shown during the COVID-19 pandemic. In this work, we report on performance of an extensive, building-by-building platform deployed across 38 locations largest private university system Mexico, spanning 19 32 states. Sampling took place weekly from January 2021 and June 2022. Data 343 sampling sites was clustered campus state evaluated through its correlation with seven-day average daily new cases each cluster. Statistically significant linear correlations (p-values below 0.05) were found 25 campuses 13 Moreover, evaluate effectiveness epidemiologic containment measures taken institution potential representative points for future emergencies Monterrey Metropolitan Area, between viral loads samples be stronger Dulces Nombres, treatment plant city (Pearson coefficient: 0.6456, p-value: 6.36710 −8 ), than study 0.4860, 8.288×10 −5 ). However, when comparing after urban mobility returned pre-pandemic levels, levels both became comparable (0.894 0.865 Nombres).

Language: Английский

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Towards geospatially-resolved public-health surveillance via wastewater sequencing

Nature Communications, Journal Year: 2024, Volume and Issue: 15(1)

Published: Sept. 27, 2024

Language: Английский

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Campus Source to Sink Wastewater Surveillance of Severe Acute Respiratory Syndrome Coronavirus-2 (SARS-CoV-2).

Current Research in Microbial Sciences, Journal Year: 2024, Volume and Issue: 6, P. 100240 - 100240

Published: Jan. 1, 2024

Wastewater-based surveillance (WBS) offers an aggregate, and cost-effective approach for tracking infectious disease outbreak prevalence within communities, that provides data on community health complementary to individual clinical testing. This study reports a 16-month WBS initiative university campus in England, UK, assessing the presence of SARS-CoV-2 sewers from large buildings, downstream sewer locations, raw wastewater, partially treated effluents. Key findings include detection Alpha (B.1.1.7) variant with 70% confirmed cases correlating positive wastewater samples. Notably, ammonium nitrogen (NH4-N) levels showed correlation (ρ = 0.543, p <0.01) virus at building scale, relationship not observed or treatment works (WWTW) due dilution. The WWTW was compliant standards, but secondary processes were efficient removal as consistently detected discharges. Tools developed through can also be used enhance traditional environmental monitoring aquatic systems. detailed source-to-sink evaluation, emphasizing critical need widespread application improvement WBS. It showcases utility reinforces ongoing challenges posed by viruses receiving water quality.

Language: Английский

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Development of biomaterial composite hydrogel as a passive sampler with potential application in wastewater-based surveillance

Heliyon, Journal Year: 2024, Volume and Issue: 10(17), P. e37014 - e37014

Published: Sept. 1, 2024

Language: Английский

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Wastewater based measures of COVID-19 and associations with children's absenteeism at grade schools

The Science of The Total Environment, Journal Year: 2024, Volume and Issue: 959, P. 178217 - 178217

Published: Dec. 27, 2024

Language: Английский

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