Development and Validation of the Skimmed Milk Pellet Extraction Protocol for SARS-CoV-2 Wastewater Surveillance.

Autor: Philo SE; Department of Environmental and Occupational Health Sciences, University of Washington, 2338 Roosevelt One Building, 4225 Roosevelt Way NE, Seattle, WA, 98105, USA., Ong AQW; Department of Environmental and Occupational Health Sciences, University of Washington, 2338 Roosevelt One Building, 4225 Roosevelt Way NE, Seattle, WA, 98105, USA., Keim EK; Department of Environmental and Occupational Health Sciences, University of Washington, 2338 Roosevelt One Building, 4225 Roosevelt Way NE, Seattle, WA, 98105, USA., Swanstrom R; Department of Environmental and Occupational Health Sciences, University of Washington, 2338 Roosevelt One Building, 4225 Roosevelt Way NE, Seattle, WA, 98105, USA., Kossik AL; Department of Environmental and Occupational Health Sciences, University of Washington, 2338 Roosevelt One Building, 4225 Roosevelt Way NE, Seattle, WA, 98105, USA., Zhou NA; Department of Environmental and Occupational Health Sciences, University of Washington, 2338 Roosevelt One Building, 4225 Roosevelt Way NE, Seattle, WA, 98105, USA., Beck NK; Department of Environmental and Occupational Health Sciences, University of Washington, 2338 Roosevelt One Building, 4225 Roosevelt Way NE, Seattle, WA, 98105, USA., Meschke JS; Department of Environmental and Occupational Health Sciences, University of Washington, 2338 Roosevelt One Building, 4225 Roosevelt Way NE, Seattle, WA, 98105, USA. jmeschke@uw.edu.
Jazyk: angličtina
Zdroj: Food and environmental virology [Food Environ Virol] 2022 Dec; Vol. 14 (4), pp. 355-363. Date of Electronic Publication: 2022 Feb 10.
DOI: 10.1007/s12560-022-09512-5
Abstrakt: Wastewater surveillance for SARS-CoV-2 may serve as a useful source of data for public health departments as the virus is shed in the stool of infected individuals. However, for wastewater data to be actionable, wastewater must be collected, concentrated, and analyzed in a timely manner. This manuscript presents modifications on a skimmed milk concentration protocol to reduce processing time, increase the number of samples that can be processed at once, and enable use in resource-limited settings. Wastewater seeded with Human coronavirus OC43 (OC43) was concentrated using a skimmed milk flocculation protocol, and then pellets were directly extracted with the QIAamp Viral RNA Mini kit. This protocol has a higher average effective volume assayed (6.35 mL) than skimmed milk concentration methods, with and without Vertrel XF™, which involve resuspension of the pellets in PBS extraction prior to nucleic acid extraction (1.28 mL, 1.44 mL, respectively). OC43 was selected as a recovery control organism because both it and SARS-CoV-2 are enveloped respiratory viruses that primarily infect humans resulting in respiratory symptoms. The OC43 percent recovery for the direct extraction protocol (3.4%) is comparable to that of skimmed milk concentration with and without Vertrel XF™ extraction (4.0%, 2.6%, respectively). When comparing SARS-CoV-2 detection using McNemar's chi-square test, the pellet extraction method is not statistically different from skimmed milk concentration, with and without Vertrel XF™ extraction. This suggests that the method performs equally as well as existing methods. Added benefits include reduced time spent per sample and the ability to process more samples at a single time. Direct extraction of skimmed milk pellets is a viable method for quick turnaround of wastewater data for public health interventions.
(© 2022. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)
Databáze: MEDLINE
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