Detection of 2019 novel coronavirus (2019-nCoV) by real-time RT-PCR

Victor M Corman, Olfert Landt, Marco Kaiser, Richard Molenkamp, Adam Meijer, Daniel Kw Chu, Tobias Bleicker, Sebastian Brünink, Julia Schneider, Marie Luisa Schmidt, Daphne Gjc Mulders, Bart L Haagmans, Bas van der Veer, Sharon van den Brink, Lisa Wijsman, Gabriel Goderski, Jean-Louis Romette, Joanna Ellis, Maria Zambon, Malik Peiris, Herman Goossens, Chantal Reusken, Marion Pg Koopmans, Christian Drosten, Victor M Corman, Olfert Landt, Marco Kaiser, Richard Molenkamp, Adam Meijer, Daniel Kw Chu, Tobias Bleicker, Sebastian Brünink, Julia Schneider, Marie Luisa Schmidt, Daphne Gjc Mulders, Bart L Haagmans, Bas van der Veer, Sharon van den Brink, Lisa Wijsman, Gabriel Goderski, Jean-Louis Romette, Joanna Ellis, Maria Zambon, Malik Peiris, Herman Goossens, Chantal Reusken, Marion Pg Koopmans, Christian Drosten

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.

Conflict of interest statement

Conflict of interest: None declared.

Figures

Figure 1
Figure 1
Relative positions of amplicon targets on the SARS coronavirus and the 2019 novel coronavirus genome
Figure 2
Figure 2
Partial alignments of oligonucleotide binding regions, SARS-related coronaviruses (n = 9)
Figure 3
Figure 3
Determination of limits of detection based on SARS coronavirus genomic RNA and 2019 novel coronavirus-specific in vitro transcribed RNA

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Source: PubMed

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