An in-well direct lysis method for rapid detection of SARS-CoV-2 by real time RT-PCR in eSwab specimens

Research output: Contribution to journalJournal articleResearchpeer-review

  • Rikke Lind Jørgensen
  • Martin Schou Pedersen
  • Alisha Shazad Chauhan
  • Louise Munkholm Andreasson
  • Gitte Qvist Kristiansen
  • Jan Gorm Lisby
  • Maiken Worsøe Rosenstierne
  • Schønning, Kristian

Background: Diagnostic real time reverse transcription PCR (rRT-PCR) is usually done using nucleic acid (NA) purified from the sample. In the SARS-CoV-2 pandemic reagents and utensils for NA purification has been in short supply. This has generated interest in methods that eliminate the need for NA purification. Objectives: To investigate if addition of detergent to rRT-PCR master mix (MM) enabled in-well direct lysis and detection of SARS-CoV-2 in clinical eSwab specimens. Study design: IGEPAL-CA-630 (IGEPAL) was added to SARS-CoV-2 MM to 0.3 % final concentration and crude sample was added directly to the PCR well containing MM. Cycle of positivity (Cp) and categorical agreement was compared in samples tested in standard rRT-PCR after NA purification and in in-well lysis, direct rRT-PCR. Results: In-well lysis direct rRT-PCR detected SARS-CoV-2 in 27/30 previously SARS-CoV-2+ samples with an average bias of 3.26 cycles (95 %CI: 0.08–6.43 cycles). All 30 previously test negative samples remained negative when tested in in-well lysis, direct PCR. Conclusions: Supplementation of detergent to MM was shown to be useful for the detection of SARS CoV-2 in eSwab specimens (COPAN) by direct rRT-PCR without prior NA purification.

Original languageEnglish
Article number114062
JournalJournal of Virological Methods
Volume289
ISSN0166-0934
DOIs
Publication statusPublished - Mar 2021

Bibliographical note

Publisher Copyright:
© 2021 Elsevier B.V.

    Research areas

  • Crude lysate, Direct PCR, IGEPAL-CA-630, SARS-CoV-2

ID: 285808786