Phenol-chloroform-based RNA purification for detection of SARS-CoV-2 by RT-qPCR: Comparison with automated systems

Henrik Dimke, Sanne L. Larsen, Marianne N. Skov, Hanne Larsen, Gitte N. Hartmeyer, Jesper B. Moeller*

*Corresponding author for this work

Research output: Contribution to journalJournal articleResearchpeer-review

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Abstract

The outbreak of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) rapidly reached pandemic levels. Sufficient testing for SARS-CoV-2 has remained essential for tracking and containing the virus. SARS-CoV-2 testing capabilities are still limited in many countries. Here, we explore the use of conventional RNA purification as an alternative to automated systems for detection of SARS-CoV-2 by RT-qPCR. 87 clinical swab specimens were extracted by conventional phenol-chloroform RNA purification and compared to commercial platforms for RNA extraction and the fully integrated Cobas®6800 diagnostic system. Our results show that the conventional RNA extraction is fully comparable to modern automated systems regarding analytical sensitivity and specificity with respect to detection of SARS-CoV-2 as evaluated by RT-qPCR. Moreover, the method is easily scalable and implemented in conventional laboratories as a low cost and suitable alternative to automated systems for the detection of SARS-CoV-2.

Original languageEnglish
Article numbere0247524
JournalPLOS ONE
Volume16
Issue number2
Number of pages6
ISSN1932-6203
DOIs
Publication statusPublished - 24. Feb 2021

Keywords

  • COVID-19 Testing/methods
  • COVID-19/diagnosis
  • Chloroform/chemistry
  • Clinical Laboratory Techniques/methods
  • Humans
  • Molecular Diagnostic Techniques/methods
  • Pandemics
  • Phenol/chemistry
  • RNA, Viral/genetics
  • RNA/genetics
  • Real-Time Polymerase Chain Reaction/methods
  • SARS-CoV-2/chemistry
  • Sensitivity and Specificity
  • Specimen Handling/methods

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