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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...
Autores principales: | , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Elsevier
2022
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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. |
format | Online Article Text |
id | pubmed-8890991 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | Elsevier |
record_format | MEDLINE/PubMed |
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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