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Sensitive extraction-free SARS-CoV-2 RNA virus detection using a chelating resin
Current conventional detection of SARS-CoV-2 involves collection of a patient’s sample with a nasopharyngeal swab, storage of the swab during transport in a viral transport medium, extraction of RNA, and quantitative reverse transcription PCR (RT-qPCR). We developed a simplified preparation method u...
Autores principales: | , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Elsevier
2021
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8349732/ https://www.ncbi.nlm.nih.gov/pubmed/34396082 http://dx.doi.org/10.1016/j.isci.2021.102960 |
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author | Guan, Bin Frank, Karen M. Maldonado, José O. Beach, Margaret Pelayo, Eileen Warner, Blake M. Hufnagel, Robert B. |
author_facet | Guan, Bin Frank, Karen M. Maldonado, José O. Beach, Margaret Pelayo, Eileen Warner, Blake M. Hufnagel, Robert B. |
author_sort | Guan, Bin |
collection | PubMed |
description | Current conventional detection of SARS-CoV-2 involves collection of a patient’s sample with a nasopharyngeal swab, storage of the swab during transport in a viral transport medium, extraction of RNA, and quantitative reverse transcription PCR (RT-qPCR). We developed a simplified preparation method using a chelating resin, Chelex, which obviates RNA extraction during viral testing. Direct detection RT-qPCR and digital droplet PCR were compared to the current conventional method with RNA extraction for simulated samples and patient specimens. The heat treatment in the presence of Chelex markedly improved detection sensitivity as compared to heat alone, and lack of RNA extraction shortens the overall diagnostic workflow. Furthermore, the initial sample heating step inactivates SARS-CoV-2 infectivity, thus improving workflow safety. This fast RNA preparation and detection method is versatile for a variety of samples, safe for testing personnel, and suitable for standard clinical collection and testing on high-throughput platforms. |
format | Online Article Text |
id | pubmed-8349732 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | Elsevier |
record_format | MEDLINE/PubMed |
spelling | pubmed-83497322021-08-09 Sensitive extraction-free SARS-CoV-2 RNA virus detection using a chelating resin Guan, Bin Frank, Karen M. Maldonado, José O. Beach, Margaret Pelayo, Eileen Warner, Blake M. Hufnagel, Robert B. iScience Article Current conventional detection of SARS-CoV-2 involves collection of a patient’s sample with a nasopharyngeal swab, storage of the swab during transport in a viral transport medium, extraction of RNA, and quantitative reverse transcription PCR (RT-qPCR). We developed a simplified preparation method using a chelating resin, Chelex, which obviates RNA extraction during viral testing. Direct detection RT-qPCR and digital droplet PCR were compared to the current conventional method with RNA extraction for simulated samples and patient specimens. The heat treatment in the presence of Chelex markedly improved detection sensitivity as compared to heat alone, and lack of RNA extraction shortens the overall diagnostic workflow. Furthermore, the initial sample heating step inactivates SARS-CoV-2 infectivity, thus improving workflow safety. This fast RNA preparation and detection method is versatile for a variety of samples, safe for testing personnel, and suitable for standard clinical collection and testing on high-throughput platforms. Elsevier 2021-08-09 /pmc/articles/PMC8349732/ /pubmed/34396082 http://dx.doi.org/10.1016/j.isci.2021.102960 Text en 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 | Article Guan, Bin Frank, Karen M. Maldonado, José O. Beach, Margaret Pelayo, Eileen Warner, Blake M. Hufnagel, Robert B. Sensitive extraction-free SARS-CoV-2 RNA virus detection using a chelating resin |
title | Sensitive extraction-free SARS-CoV-2 RNA virus detection using a chelating resin |
title_full | Sensitive extraction-free SARS-CoV-2 RNA virus detection using a chelating resin |
title_fullStr | Sensitive extraction-free SARS-CoV-2 RNA virus detection using a chelating resin |
title_full_unstemmed | Sensitive extraction-free SARS-CoV-2 RNA virus detection using a chelating resin |
title_short | Sensitive extraction-free SARS-CoV-2 RNA virus detection using a chelating resin |
title_sort | sensitive extraction-free sars-cov-2 rna virus detection using a chelating resin |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8349732/ https://www.ncbi.nlm.nih.gov/pubmed/34396082 http://dx.doi.org/10.1016/j.isci.2021.102960 |
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