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Direct RT-PCR amplification of SARS-CoV-2 from clinical samples using a concentrated viral lysis-amplification buffer prepared with IGEPAL-630

The pandemic of 2019 caused by the novel coronavirus (SARS-CoV-2) is still rapidly spreading worldwide. Nucleic acid amplification serves as the gold standard method for confirmation of COVID-19 infection. However, challenges faced for diagnostic laboratories from undeveloped countries includes shor...

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Autores principales: Castellanos-Gonzalez, Alejandro, Shelite, Thomas R., Lloyd, Nicole, Sadiqova, Aygul, Ping, Ren, Williams-Bouyer, Natalie, Melby, Peter C., Travi, Bruno L.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8270935/
https://www.ncbi.nlm.nih.gov/pubmed/34244543
http://dx.doi.org/10.1038/s41598-021-93333-2
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author Castellanos-Gonzalez, Alejandro
Shelite, Thomas R.
Lloyd, Nicole
Sadiqova, Aygul
Ping, Ren
Williams-Bouyer, Natalie
Melby, Peter C.
Travi, Bruno L.
author_facet Castellanos-Gonzalez, Alejandro
Shelite, Thomas R.
Lloyd, Nicole
Sadiqova, Aygul
Ping, Ren
Williams-Bouyer, Natalie
Melby, Peter C.
Travi, Bruno L.
author_sort Castellanos-Gonzalez, Alejandro
collection PubMed
description The pandemic of 2019 caused by the novel coronavirus (SARS-CoV-2) is still rapidly spreading worldwide. Nucleic acid amplification serves as the gold standard method for confirmation of COVID-19 infection. However, challenges faced for diagnostic laboratories from undeveloped countries includes shortage of kits and supplies to purify viral RNA. Therefore, it is urgent to validate alternative nucleic acid isolation methods for SARS-CoV-2. Our results demonstrate that a concentrated viral lysis amplification buffer (vLAB) prepared with the nonionic detergent IGEPAL enables qualitative detection of SARS-CoV-2 by direct Reverse Transcriptase-Polymerase Chain Reaction (dRT-PCR). Furthermore, vLAB was effective in inactivating SARS-CoV-2. Since this method is inexpensive and no RNA purification equipment or additional cDNA synthesis is required, this dRT-PCR with vLAB should be considered as an alternative method for qualitative detection of SARS-CoV-2.
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spelling pubmed-82709352021-07-12 Direct RT-PCR amplification of SARS-CoV-2 from clinical samples using a concentrated viral lysis-amplification buffer prepared with IGEPAL-630 Castellanos-Gonzalez, Alejandro Shelite, Thomas R. Lloyd, Nicole Sadiqova, Aygul Ping, Ren Williams-Bouyer, Natalie Melby, Peter C. Travi, Bruno L. Sci Rep Article The pandemic of 2019 caused by the novel coronavirus (SARS-CoV-2) is still rapidly spreading worldwide. Nucleic acid amplification serves as the gold standard method for confirmation of COVID-19 infection. However, challenges faced for diagnostic laboratories from undeveloped countries includes shortage of kits and supplies to purify viral RNA. Therefore, it is urgent to validate alternative nucleic acid isolation methods for SARS-CoV-2. Our results demonstrate that a concentrated viral lysis amplification buffer (vLAB) prepared with the nonionic detergent IGEPAL enables qualitative detection of SARS-CoV-2 by direct Reverse Transcriptase-Polymerase Chain Reaction (dRT-PCR). Furthermore, vLAB was effective in inactivating SARS-CoV-2. Since this method is inexpensive and no RNA purification equipment or additional cDNA synthesis is required, this dRT-PCR with vLAB should be considered as an alternative method for qualitative detection of SARS-CoV-2. Nature Publishing Group UK 2021-07-09 /pmc/articles/PMC8270935/ /pubmed/34244543 http://dx.doi.org/10.1038/s41598-021-93333-2 Text en © The Author(s) 2021 https://creativecommons.org/licenses/by/4.0/Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) .
spellingShingle Article
Castellanos-Gonzalez, Alejandro
Shelite, Thomas R.
Lloyd, Nicole
Sadiqova, Aygul
Ping, Ren
Williams-Bouyer, Natalie
Melby, Peter C.
Travi, Bruno L.
Direct RT-PCR amplification of SARS-CoV-2 from clinical samples using a concentrated viral lysis-amplification buffer prepared with IGEPAL-630
title Direct RT-PCR amplification of SARS-CoV-2 from clinical samples using a concentrated viral lysis-amplification buffer prepared with IGEPAL-630
title_full Direct RT-PCR amplification of SARS-CoV-2 from clinical samples using a concentrated viral lysis-amplification buffer prepared with IGEPAL-630
title_fullStr Direct RT-PCR amplification of SARS-CoV-2 from clinical samples using a concentrated viral lysis-amplification buffer prepared with IGEPAL-630
title_full_unstemmed Direct RT-PCR amplification of SARS-CoV-2 from clinical samples using a concentrated viral lysis-amplification buffer prepared with IGEPAL-630
title_short Direct RT-PCR amplification of SARS-CoV-2 from clinical samples using a concentrated viral lysis-amplification buffer prepared with IGEPAL-630
title_sort direct rt-pcr amplification of sars-cov-2 from clinical samples using a concentrated viral lysis-amplification buffer prepared with igepal-630
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8270935/
https://www.ncbi.nlm.nih.gov/pubmed/34244543
http://dx.doi.org/10.1038/s41598-021-93333-2
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