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Homebrew: An economical and sensitive glassmilk-based nucleic-acid extraction method for SARS-CoV-2 diagnostics

Management of COVID-19 and other epidemics requires large-scale diagnostic testing. The gold standard for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection remains reverse transcription quantitative PCR (qRT-PCR) analysis, which detects viral RNA more sensitively than any other...

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Autores principales: Page, Robert, Scourfield, Edward, Ficarelli, Mattia, McKellar, Stuart W., Lee, Kwok Leung, Maguire, Thomas J.A., Bouton, Clement, Lista, Maria Jose, Neil, Stuart J.D., Malim, Michael H., Zuckerman, Mark, Mischo, Hannah E., Martinez-Nunez, Rocio T.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8890991/
https://www.ncbi.nlm.nih.gov/pubmed/35262039
http://dx.doi.org/10.1016/j.crmeth.2022.100186
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author Page, Robert
Scourfield, Edward
Ficarelli, Mattia
McKellar, Stuart W.
Lee, Kwok Leung
Maguire, Thomas J.A.
Bouton, Clement
Lista, Maria Jose
Neil, Stuart J.D.
Malim, Michael H.
Zuckerman, Mark
Mischo, Hannah E.
Martinez-Nunez, Rocio T.
author_facet Page, Robert
Scourfield, Edward
Ficarelli, Mattia
McKellar, Stuart W.
Lee, Kwok Leung
Maguire, Thomas J.A.
Bouton, Clement
Lista, Maria Jose
Neil, Stuart J.D.
Malim, Michael H.
Zuckerman, Mark
Mischo, Hannah E.
Martinez-Nunez, Rocio T.
author_sort Page, Robert
collection PubMed
description Management of COVID-19 and other epidemics requires large-scale diagnostic testing. The gold standard for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection remains reverse transcription quantitative PCR (qRT-PCR) analysis, which detects viral RNA more sensitively than any other method. However, the resource use and supply-chain requirements of RT-PCR have continued to challenge diagnostic laboratories worldwide. Here, we establish and characterize a low-cost method to detect SARS-CoV-2 in clinical combined nose and throat swabs, allowing for automation in high-throughput settings. This method inactivates virus material with sodium dodecylsulfate (SDS) and uses silicon dioxide as the RNA-binding matrix in combination with sodium chloride (NaCl) and isopropanol. With similar sensitivity for SARS-CoV-2 viral targets but a fraction of time and reagent expenditure compared with commercial kits, our method also enables sample pooling without loss of sensitivity. We suggest that this method will facilitate more economical widespread testing, particularly in resource-limited settings.
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spelling pubmed-88909912022-03-04 Homebrew: An economical and sensitive glassmilk-based nucleic-acid extraction method for SARS-CoV-2 diagnostics Page, Robert Scourfield, Edward Ficarelli, Mattia McKellar, Stuart W. Lee, Kwok Leung Maguire, Thomas J.A. Bouton, Clement Lista, Maria Jose Neil, Stuart J.D. Malim, Michael H. Zuckerman, Mark Mischo, Hannah E. Martinez-Nunez, Rocio T. Cell Rep Methods Report Management of COVID-19 and other epidemics requires large-scale diagnostic testing. The gold standard for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection remains reverse transcription quantitative PCR (qRT-PCR) analysis, which detects viral RNA more sensitively than any other method. However, the resource use and supply-chain requirements of RT-PCR have continued to challenge diagnostic laboratories worldwide. Here, we establish and characterize a low-cost method to detect SARS-CoV-2 in clinical combined nose and throat swabs, allowing for automation in high-throughput settings. This method inactivates virus material with sodium dodecylsulfate (SDS) and uses silicon dioxide as the RNA-binding matrix in combination with sodium chloride (NaCl) and isopropanol. With similar sensitivity for SARS-CoV-2 viral targets but a fraction of time and reagent expenditure compared with commercial kits, our method also enables sample pooling without loss of sensitivity. We suggest that this method will facilitate more economical widespread testing, particularly in resource-limited settings. Elsevier 2022-03-03 /pmc/articles/PMC8890991/ /pubmed/35262039 http://dx.doi.org/10.1016/j.crmeth.2022.100186 Text en © 2022 The Authors https://creativecommons.org/licenses/by/4.0/This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Report
Page, Robert
Scourfield, Edward
Ficarelli, Mattia
McKellar, Stuart W.
Lee, Kwok Leung
Maguire, Thomas J.A.
Bouton, Clement
Lista, Maria Jose
Neil, Stuart J.D.
Malim, Michael H.
Zuckerman, Mark
Mischo, Hannah E.
Martinez-Nunez, Rocio T.
Homebrew: An economical and sensitive glassmilk-based nucleic-acid extraction method for SARS-CoV-2 diagnostics
title Homebrew: An economical and sensitive glassmilk-based nucleic-acid extraction method for SARS-CoV-2 diagnostics
title_full Homebrew: An economical and sensitive glassmilk-based nucleic-acid extraction method for SARS-CoV-2 diagnostics
title_fullStr Homebrew: An economical and sensitive glassmilk-based nucleic-acid extraction method for SARS-CoV-2 diagnostics
title_full_unstemmed Homebrew: An economical and sensitive glassmilk-based nucleic-acid extraction method for SARS-CoV-2 diagnostics
title_short Homebrew: An economical and sensitive glassmilk-based nucleic-acid extraction method for SARS-CoV-2 diagnostics
title_sort homebrew: an economical and sensitive glassmilk-based nucleic-acid extraction method for sars-cov-2 diagnostics
topic Report
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8890991/
https://www.ncbi.nlm.nih.gov/pubmed/35262039
http://dx.doi.org/10.1016/j.crmeth.2022.100186
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