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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...

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Detalles Bibliográficos
Autores principales: Guan, Bin, Frank, Karen M., Maldonado, José O., Beach, Margaret, Pelayo, Eileen, Warner, Blake M., Hufnagel, Robert B.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2021
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.
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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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