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A Fast and Accessible Method for the Isolation of RNA, DNA, and Protein To Facilitate the Detection of SARS-CoV-2
Management of the coronavirus disease 2019 (COVID-19) pandemic requires widespread testing for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). A main limitation for widespread SARS-CoV-2 testing is the global shortage of essential supplies, among them RNA extraction kits. The need for...
Autores principales: | , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
American Society for Microbiology
2021
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8092744/ https://www.ncbi.nlm.nih.gov/pubmed/33293367 http://dx.doi.org/10.1128/JCM.02403-20 |
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author | Ponce-Rojas, Jose Carlos Costello, Michael S. Proctor, Duncan A. Kosik, Kenneth S. Wilson, Maxwell Z. Arias, Carolina Acosta-Alvear, Diego |
author_facet | Ponce-Rojas, Jose Carlos Costello, Michael S. Proctor, Duncan A. Kosik, Kenneth S. Wilson, Maxwell Z. Arias, Carolina Acosta-Alvear, Diego |
author_sort | Ponce-Rojas, Jose Carlos |
collection | PubMed |
description | Management of the coronavirus disease 2019 (COVID-19) pandemic requires widespread testing for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). A main limitation for widespread SARS-CoV-2 testing is the global shortage of essential supplies, among them RNA extraction kits. The need for commercial RNA extraction kits places a bottleneck on tests that detect SARS-CoV-2 genetic material, including PCR-based reference tests. Here, we propose an alternative method we call PEARL (precipitation-enhanced analyte retrieval) that addresses this limitation. PEARL uses a lysis solution that disrupts cell membranes and viral envelopes while simultaneously providing conditions suitable for alcohol-based precipitation of RNA, DNA, and proteins. PEARL is a fast, low-cost, and simple method that uses common laboratory reagents and offers performance comparable to that of commercial RNA extraction kits. PEARL offers an alternative method to isolate host and pathogen nucleic acids and proteins to streamline the detection of DNA and RNA viruses, including SARS-CoV-2. |
format | Online Article Text |
id | pubmed-8092744 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | American Society for Microbiology |
record_format | MEDLINE/PubMed |
spelling | pubmed-80927442021-05-21 A Fast and Accessible Method for the Isolation of RNA, DNA, and Protein To Facilitate the Detection of SARS-CoV-2 Ponce-Rojas, Jose Carlos Costello, Michael S. Proctor, Duncan A. Kosik, Kenneth S. Wilson, Maxwell Z. Arias, Carolina Acosta-Alvear, Diego J Clin Microbiol Virology Management of the coronavirus disease 2019 (COVID-19) pandemic requires widespread testing for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). A main limitation for widespread SARS-CoV-2 testing is the global shortage of essential supplies, among them RNA extraction kits. The need for commercial RNA extraction kits places a bottleneck on tests that detect SARS-CoV-2 genetic material, including PCR-based reference tests. Here, we propose an alternative method we call PEARL (precipitation-enhanced analyte retrieval) that addresses this limitation. PEARL uses a lysis solution that disrupts cell membranes and viral envelopes while simultaneously providing conditions suitable for alcohol-based precipitation of RNA, DNA, and proteins. PEARL is a fast, low-cost, and simple method that uses common laboratory reagents and offers performance comparable to that of commercial RNA extraction kits. PEARL offers an alternative method to isolate host and pathogen nucleic acids and proteins to streamline the detection of DNA and RNA viruses, including SARS-CoV-2. American Society for Microbiology 2021-03-19 /pmc/articles/PMC8092744/ /pubmed/33293367 http://dx.doi.org/10.1128/JCM.02403-20 Text en Copyright © 2021 Ponce-Rojas et al. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International license (https://creativecommons.org/licenses/by/4.0/) . https://doi.org/10.1128/ASMCopyrightv2This article is made available via the PMC Open Access Subset for unrestricted noncommercial re-use and secondary analysis in any form or by any means with acknowledgement of the original source. These permissions are granted for the duration of the World Health Organization (WHO) declaration of COVID-19 as a global pandemic. |
spellingShingle | Virology Ponce-Rojas, Jose Carlos Costello, Michael S. Proctor, Duncan A. Kosik, Kenneth S. Wilson, Maxwell Z. Arias, Carolina Acosta-Alvear, Diego A Fast and Accessible Method for the Isolation of RNA, DNA, and Protein To Facilitate the Detection of SARS-CoV-2 |
title | A Fast and Accessible Method for the Isolation of RNA, DNA, and Protein To Facilitate the Detection of SARS-CoV-2 |
title_full | A Fast and Accessible Method for the Isolation of RNA, DNA, and Protein To Facilitate the Detection of SARS-CoV-2 |
title_fullStr | A Fast and Accessible Method for the Isolation of RNA, DNA, and Protein To Facilitate the Detection of SARS-CoV-2 |
title_full_unstemmed | A Fast and Accessible Method for the Isolation of RNA, DNA, and Protein To Facilitate the Detection of SARS-CoV-2 |
title_short | A Fast and Accessible Method for the Isolation of RNA, DNA, and Protein To Facilitate the Detection of SARS-CoV-2 |
title_sort | fast and accessible method for the isolation of rna, dna, and protein to facilitate the detection of sars-cov-2 |
topic | Virology |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8092744/ https://www.ncbi.nlm.nih.gov/pubmed/33293367 http://dx.doi.org/10.1128/JCM.02403-20 |
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