SARS-CoV-2 detection by direct rRT-PCR without RNA extraction

J Clin Virol. 2020 Jul:128:104423. doi: 10.1016/j.jcv.2020.104423. Epub 2020 May 7.

Abstract

Rapid and reliable screening of SARS-CoV-2 is fundamental to assess viral spread and limit the pandemic we are facing. In this study, we compared direct rRT-PCR method (without RNA extraction) using SeeGene AllplexTM 2019-nCoV rRT-PCR with the RealStar® SARS-CoV-2 rRT-PCR kit (Altona Diagnostics). Furthermore, we assessed the impact of swab storage media composition on PCR efficiency. We show that SeeGene and Altona's assays provide similar efficiency. Importantly, we provide evidence that RNA extraction can be successfully bypassed when samples are stored in UTM medium or in molecular water but not when samples are stored in saline solution and in Hanks medium.

Keywords: COVID19; Coronavirus; Direct rRTPCR; RNA extraction; SARS-CoV-2; Virus detection.

MeSH terms

  • Betacoronavirus / genetics
  • Betacoronavirus / isolation & purification*
  • COVID-19
  • Coronavirus Infections / diagnosis*
  • Coronavirus Infections / virology
  • Humans
  • Nasopharynx / virology
  • Pandemics
  • Pneumonia, Viral / diagnosis*
  • Pneumonia, Viral / virology
  • RNA, Viral / genetics
  • Reverse Transcriptase Polymerase Chain Reaction / methods*
  • SARS-CoV-2
  • Specimen Handling*

Substances

  • RNA, Viral