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Expanding a Wastewater-Based Surveillance Methodology for DNA Isolation from a Workflow Optimized for SARS-CoV-2 RNA Quantification.
Babler, Kristina M; Sharkey, Mark E; Amirali, Ayaaz; Boone, Melinda M; Comerford, Samuel; Currall, Benjamin B; Grills, George S; Laine, Jennifer; Mason, Christopher E; Reding, Brian; Schürer, Stephan; Stevenson, Mario; Vidovic, Dusica; Williams, Sion L; Solo-Gabriele, Helena M.
Afiliação
  • Babler KM; Department of Chemical Environmental and Materials Engineering University of Miami Coral GablesFlorida33124 USA.
  • Sharkey ME; Department of Medicine University of Miami Miller School of Medicine MiamiFlorida33136 USA.
  • Amirali A; Department of Chemical Environmental and Materials Engineering University of Miami Coral GablesFlorida33124 USA.
  • Boone MM; Sylvester Comprehensive Cancer Center University of Miami Miller School of Medicine MiamiFlorida33136 USA.
  • Comerford S; Department of Medicine University of Miami Miller School of Medicine MiamiFlorida33136 USA.
  • Currall BB; Sylvester Comprehensive Cancer Center University of Miami Miller School of Medicine MiamiFlorida33136 USA.
  • Grills GS; Sylvester Comprehensive Cancer Center University of Miami Miller School of Medicine MiamiFlorida33136 USA.
  • Laine J; Environmental Health and Safety University of Miami MiamiFlorida33136 USA.
  • Mason CE; Department of Physiology and Biophysics Weill Cornell Medicine New YorkNew York10065 USA.
  • Reding B; The HRH Prince Alwaleed Bin Talal Bin Abdulaziz Al-Saud Institute for Computational Biomedicine Weill Cornell Medicine New YorkNew York10065 USA.
  • Schürer S; The WorldQuant Initiative for Quantitative Prediction Weill Cornell Medicine New YorkNew YorkUSA 10065 USA.
  • Stevenson M; Environmental Health and Safety University of Miami MiamiFlorida33136 USA.
  • Vidovic D; Sylvester Comprehensive Cancer Center University of Miami Miller School of Medicine MiamiFlorida33136 USA.
  • Williams SL; Department of Molecular and Cellular Pharmacology University of Miami Miller School of Medicine MiamiFlorida33136 USA.
  • Solo-Gabriele HM; Institute for Data Science & Computing, University of Miami Coral GablesFlorida33146 USA.
J Biomol Tech ; 34(4)2023 Dec.
Article em En | MEDLINE | ID: mdl-38268997
ABSTRACT
Wastewater-based surveillance (WBS) is a noninvasive, epidemiological strategy for assessing the spread of COVID-19 in communities. This strategy was based upon wastewater RNA measurements of the viral target, severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2). The utility of WBS for assessing the spread of COVID-19 has motivated research to measure targets beyond SARS-CoV-2, including pathogens containing DNA. The objective of this study was to establish the necessary steps for isolating DNA from wastewater by modifying a long-standing RNA-specific extraction workflow optimized for SARS-CoV-2 detection. Modifications were made to the sample concentration process and included an evaluation of bead bashing prior to the extraction of either DNA or RNA. Results showed that bead bashing reduced detection of RNA from wastewater but improved recovery of DNA as assessed by quantitative polymerase chain reaction (qPCR). Bead bashing is therefore not recommended for the quantification of RNA viruses using qPCR. Whereas for Mycobacterium bacterial DNA isolation, bead bashing was necessary for improving qPCR quantification. Overall, we recommend 2 separate workflows, one for RNA viruses that does not include bead bashing and one for other microbes that use bead bashing for DNA isolation. The experimentation done here shows that current-standing WBS program methodologies optimized for SARS-CoV-2 need to be modified and reoptimized to allow for alternative pathogens to be readily detected and monitored, expanding its utility as a tool for public health assessment.
Assuntos

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: COVID-19 Tipo de estudo: Screening_studies Limite: Humans Idioma: En Revista: J Biomol Tech Assunto da revista: BIOTECNOLOGIA Ano de publicação: 2023 Tipo de documento: Article

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: COVID-19 Tipo de estudo: Screening_studies Limite: Humans Idioma: En Revista: J Biomol Tech Assunto da revista: BIOTECNOLOGIA Ano de publicação: 2023 Tipo de documento: Article