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Proteinase K treatment in absence of RNA isolation classical procedures is a quick and cheaper alternative for SARS-CoV-2 molecular detection
The World Health Organization (WHO) has declared a pandemic of COVID-19, the disease caused by the recently described SARS-CoV-2. The relevance and importance of mass diagnosis in order to find the asymptomatic individuals is widely recognized as a mandatory tool to reinforce the control measures fo...
Autores principales: | , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Elsevier B.V.
2021
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8008826/ https://www.ncbi.nlm.nih.gov/pubmed/33798606 http://dx.doi.org/10.1016/j.jviromet.2021.114131 |
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author | Mallmann, L. Hermann, B.S. Schallenberger, K. Demoliner, M. Eisen, A.K.A. Heldt, F.H. Gularte, J.S. Hansen, A.W. de Almeida, P.R. Weber, M.N. Spilki, F.R. Fleck, J.D. |
author_facet | Mallmann, L. Hermann, B.S. Schallenberger, K. Demoliner, M. Eisen, A.K.A. Heldt, F.H. Gularte, J.S. Hansen, A.W. de Almeida, P.R. Weber, M.N. Spilki, F.R. Fleck, J.D. |
author_sort | Mallmann, L. |
collection | PubMed |
description | The World Health Organization (WHO) has declared a pandemic of COVID-19, the disease caused by the recently described SARS-CoV-2. The relevance and importance of mass diagnosis in order to find the asymptomatic individuals is widely recognized as a mandatory tool to reinforce the control measures for monitoring virus circulation and reduce the spreading of SARS-CoV-2. Here, we described quickness and cheaper strategies of direct RT-qPCR (in the absence of RNA isolation) and compared the results to those obtained using standard RNA isolation procedure. The tests varied using pure, diluted samples, combined with Proteinase K (PK) or Lysis Buffer. Our findings showed consistently that PK pre-treated samples in the absence of RNA extraction procedures presents similar results to those obtained by standard RNA isolation procedures. On average, 16 samples extracted with the MagMAX™ CORE Kit, take around 2 h, costing an average of USD 5, the pre-treatment of samples using PK, on the other hand, would cut the value to less than USD 0.30 and reduce the time of procedure in more than 1 ½ hours. The present study suggests the use of PK treatment instead of RNA isolation in order to reduce costs and time in processing samples for molecular diagnosis of SARS-CoV-2. |
format | Online Article Text |
id | pubmed-8008826 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | Elsevier B.V. |
record_format | MEDLINE/PubMed |
spelling | pubmed-80088262021-03-31 Proteinase K treatment in absence of RNA isolation classical procedures is a quick and cheaper alternative for SARS-CoV-2 molecular detection Mallmann, L. Hermann, B.S. Schallenberger, K. Demoliner, M. Eisen, A.K.A. Heldt, F.H. Gularte, J.S. Hansen, A.W. de Almeida, P.R. Weber, M.N. Spilki, F.R. Fleck, J.D. J Virol Methods Short Communication The World Health Organization (WHO) has declared a pandemic of COVID-19, the disease caused by the recently described SARS-CoV-2. The relevance and importance of mass diagnosis in order to find the asymptomatic individuals is widely recognized as a mandatory tool to reinforce the control measures for monitoring virus circulation and reduce the spreading of SARS-CoV-2. Here, we described quickness and cheaper strategies of direct RT-qPCR (in the absence of RNA isolation) and compared the results to those obtained using standard RNA isolation procedure. The tests varied using pure, diluted samples, combined with Proteinase K (PK) or Lysis Buffer. Our findings showed consistently that PK pre-treated samples in the absence of RNA extraction procedures presents similar results to those obtained by standard RNA isolation procedures. On average, 16 samples extracted with the MagMAX™ CORE Kit, take around 2 h, costing an average of USD 5, the pre-treatment of samples using PK, on the other hand, would cut the value to less than USD 0.30 and reduce the time of procedure in more than 1 ½ hours. The present study suggests the use of PK treatment instead of RNA isolation in order to reduce costs and time in processing samples for molecular diagnosis of SARS-CoV-2. Elsevier B.V. 2021-07 2021-03-30 /pmc/articles/PMC8008826/ /pubmed/33798606 http://dx.doi.org/10.1016/j.jviromet.2021.114131 Text en © 2021 Elsevier B.V. Since January 2020 Elsevier has created a COVID-19 resource centre with free information in English and Mandarin on the novel coronavirus COVID-19. The COVID-19 resource centre is hosted on Elsevier Connect, the company's public news and information website. Elsevier hereby grants permission to make all its COVID-19-related research that is available on the COVID-19 resource centre - including this research content - immediately available in PubMed Central and other publicly funded repositories, such as the WHO COVID database with rights for unrestricted research re-use and analyses in any form or by any means with acknowledgement of the original source. These permissions are granted for free by Elsevier for as long as the COVID-19 resource centre remains active. |
spellingShingle | Short Communication Mallmann, L. Hermann, B.S. Schallenberger, K. Demoliner, M. Eisen, A.K.A. Heldt, F.H. Gularte, J.S. Hansen, A.W. de Almeida, P.R. Weber, M.N. Spilki, F.R. Fleck, J.D. Proteinase K treatment in absence of RNA isolation classical procedures is a quick and cheaper alternative for SARS-CoV-2 molecular detection |
title | Proteinase K treatment in absence of RNA isolation classical procedures is a quick and cheaper alternative for SARS-CoV-2 molecular detection |
title_full | Proteinase K treatment in absence of RNA isolation classical procedures is a quick and cheaper alternative for SARS-CoV-2 molecular detection |
title_fullStr | Proteinase K treatment in absence of RNA isolation classical procedures is a quick and cheaper alternative for SARS-CoV-2 molecular detection |
title_full_unstemmed | Proteinase K treatment in absence of RNA isolation classical procedures is a quick and cheaper alternative for SARS-CoV-2 molecular detection |
title_short | Proteinase K treatment in absence of RNA isolation classical procedures is a quick and cheaper alternative for SARS-CoV-2 molecular detection |
title_sort | proteinase k treatment in absence of rna isolation classical procedures is a quick and cheaper alternative for sars-cov-2 molecular detection |
topic | Short Communication |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8008826/ https://www.ncbi.nlm.nih.gov/pubmed/33798606 http://dx.doi.org/10.1016/j.jviromet.2021.114131 |
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