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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.
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.
  • Mallmann L; Laboratório De Microbiologia Molecular, Universidade Feevale, Novo Hamburgo, Brazil.
  • Hermann BS; Laboratório De Microbiologia Molecular, Universidade Feevale, Novo Hamburgo, Brazil.
  • Schallenberger K; Laboratório De Microbiologia Molecular, Universidade Feevale, Novo Hamburgo, Brazil.
  • Demoliner M; Laboratório De Microbiologia Molecular, Universidade Feevale, Novo Hamburgo, Brazil.
  • Eisen AKA; Laboratório De Microbiologia Molecular, Universidade Feevale, Novo Hamburgo, Brazil.
  • Heldt FH; Laboratório De Microbiologia Molecular, Universidade Feevale, Novo Hamburgo, Brazil.
  • Gularte JS; Laboratório De Microbiologia Molecular, Universidade Feevale, Novo Hamburgo, Brazil.
  • Hansen AW; Laboratório De Microbiologia Molecular, Universidade Feevale, Novo Hamburgo, Brazil.
  • de Almeida PR; Laboratório De Microbiologia Molecular, Universidade Feevale, Novo Hamburgo, Brazil.
  • Weber MN; Laboratório De Microbiologia Molecular, Universidade Feevale, Novo Hamburgo, Brazil.
  • Spilki FR; Laboratório De Microbiologia Molecular, Universidade Feevale, Novo Hamburgo, Brazil.
  • Fleck JD; Laboratório De Microbiologia Molecular, Universidade Feevale, Novo Hamburgo, Brazil. Electronic address: julianefleck@feevale.br.
J Virol Methods ; 293: 114131, 2021 07.
Article in English | MEDLINE | ID: covidwho-1157565
ABSTRACT
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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Full text: Available Collection: International databases Database: MEDLINE Main subject: RNA, Viral / Endopeptidase K / COVID-19 Nucleic Acid Testing / SARS-CoV-2 / COVID-19 Type of study: Diagnostic study Limits: Humans Language: English Journal: J Virol Methods Year: 2021 Document Type: Article Affiliation country: J.jviromet.2021.114131

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Full text: Available Collection: International databases Database: MEDLINE Main subject: RNA, Viral / Endopeptidase K / COVID-19 Nucleic Acid Testing / SARS-CoV-2 / COVID-19 Type of study: Diagnostic study Limits: Humans Language: English Journal: J Virol Methods Year: 2021 Document Type: Article Affiliation country: J.jviromet.2021.114131