Comparison of commercial realtime reverse transcription PCR assays for the detection of SARS-CoV-2

Zsófia Iglói, Margareta Leven, Zain Abdel-Karem Abou-Nouar, Babette Weller, Veerle Matheeussen, Jasmine Coppens, Marion Koopmans, Richard Molenkamp

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Abstract

The emergence of a new coronavirus in Wuhan China has triggered a global need for accurate diagnostic assays. Initially, mostly laboratory developed molecular tests were available but shortly thereafter different commercial assays started to appear and are still increasing in number. Although independent performance evaluations are ongoing, available data is still scarce. Here we provide a direct comparison of key performance characteristics of 13 commercial RT-PCR assays. Thirteen RT-PCR assays were selected based on the criteria that they can be used following generic RNA extraction protocols, on common PCR platforms and availability. Using a 10-fold and 2-fold dilution series of a quantified SARS-CoV-2 cell-cultured virus stock, performance was assessed compared to our in house validated assay. Specificity was tested by using RNA extracted from cultured common human coronaviruses. All RT-PCR kits included in this study exhibited PCR efficiencies > 90%, except for the Sentinel Diagnostics B E-gene RUO assay (80%). Analytical sensitivity varied between 3.3 RNA copies to 330 RNA copies. Only one assay cross reacted with another human coronavirus (MERS). This study provides a technical baseline of 13 different commercial PCR assays for SARS-CoV-2 detection that can be used by laboratories interested in purchasing any of these for further full clinical validation.

Original languageEnglish
Article number104510
JournalJournal of Clinical Virology
Volume129
ISSN1386-6532
DOIs
Publication statusPublished - 2020

Keywords

  • Betacoronavirus/genetics
  • COVID-19
  • COVID-19 Testing
  • Clinical Laboratory Techniques/methods
  • Coronavirus Infections/diagnosis
  • Cross Reactions
  • Humans
  • Molecular Diagnostic Techniques/methods
  • Pandemics
  • Pneumonia, Viral/diagnosis
  • RNA, Viral/analysis
  • Reverse Transcriptase Polymerase Chain Reaction/methods
  • SARS-CoV-2
  • Sensitivity and Specificity

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