Detection of 2019 Novel Coronavirus (2019-nCoV) by Real-Time RT-PCR

Euro Surveill. 2020 Jan;25(3):2000045. doi: 10.2807/1560-7917.ES.2020.25.3.2000045.

Abstract

Background: The ongoing outbreak of the recently emerged novel coronavirus (2019-nCoV) poses a challenge for public health laboratories as virus isolates are unavailable while there is growing evidence that the outbreak is more widespread than initially thought, and international spread through travellers does already occur.

Aim: We aimed to develop and deploy robust diagnostic methodology for use in public health laboratory settings without having virus material available.

Methods: Here we present a validated diagnostic workflow for 2019-nCoV, its design relying on close genetic relatedness of 2019-nCoV with SARS coronavirus, making use of synthetic nucleic acid technology.

Results: The workflow reliably detects 2019-nCoV, and further discriminates 2019-nCoV from SARS-CoV. Through coordination between academic and public laboratories, we confirmed assay exclusivity based on 297 original clinical specimens containing a full spectrum of human respiratory viruses. Control material is made available through European Virus Archive - Global (EVAg), a European Union infrastructure project.

Conclusion: The present study demonstrates the enormous response capacity achieved through coordination of academic and public laboratories in national and European research networks.

Keywords: 2019-nCoV; RT-PCR; Wuhan; diagnostics; laboratory; novel coronavirus; outbreak; testing.

MeSH terms

  • Clinical Laboratory Techniques* / methods
  • Coronavirus / classification*
  • Coronavirus / genetics*
  • Coronavirus / isolation & purification
  • Coronavirus Infections / diagnosis*
  • Coronavirus Infections / virology*
  • Disease Outbreaks
  • Humans
  • RNA, Viral / analysis
  • Real-Time Polymerase Chain Reaction / methods
  • Sensitivity and Specificity

Substances

  • RNA, Viral

Supplementary concepts

  • COVID-19 diagnostic testing