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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...

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Autores principales: 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.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier B.V. 2021
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.
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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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