Detection of a novel human coronavirus by real-time reverse-transcription polymerase chain reaction

V M Corman; I Eckerle; T Bleicker; A Zaki; O Landt; M Eschbach-Bludau; S van Boheemen; R Gopal; M Ballhause; T M Bestebroer; D Muth; M A Müller; J F Drexler; M Zambon; A D Osterhaus; R M Fouchier; C Drosten

doi:10.2807/ese.17.39.20285-en

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Detection of a novel human coronavirus by real-time reverse-transcription polymerase chain reaction
V M Corman^1,2 , I Eckerle¹ , T Bleicker¹ , A Zaki³ , O Landt⁴ , M Eschbach-Bludau¹ , S van Boheemen⁵ , R Gopal⁶ , M Ballhause⁴ , T M Bestebroer⁵ , D Muth¹ , M A Müller¹ , J F Drexler¹ , M Zambon⁶ , A D Osterhaus⁵ , R M Fouchier⁵ , C Drosten¹
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Affiliations: ¹ Institute of Virology, University of Bonn Medical Centre, Bonn, Germany ² German Centre for Infection Research (DZIF), Germany ³ Virology Laboratory, Dr Soliman Fakeeh Hospital, Jeddah ⁴ TibMolbiol, Berlin, Germany ⁵ Department of Virology and Virosciences, Erasmus Medical Centre, Rotterdam, The Netherlands ⁶ Health Protection Agency (HPA), London, United Kingdom

Correspondence:
C Drosten
drosten virology-bonn.de
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Citation style for this article: Corman V M, Eckerle I, Bleicker T, Zaki A, Landt O, Eschbach-Bludau M, van Boheemen S, Gopal R, Ballhause M, Bestebroer T M, Muth D, Müller M A, Drexler J F, Zambon M, Osterhaus A D, Fouchier R M, Drosten C. Detection of a novel human coronavirus by real-time reverse-transcription polymerase chain reaction. Euro Surveill. 2012;17(39):pii=20285. https://doi.org/10.2807/ese.17.39.20285-en Received: 27 Sept 2012

Abstract

We present two real-time reverse-transcription polymerase chain reaction assays for a novel human coronavirus (CoV), targeting regions upstream of the E gene (upE) or within open reading frame (ORF)1b, respectively. Sensitivity for upE is 3.4 copies per reaction (95% confidence interval (CI): 2.5-6.9 copies) or 291 copies/mL of sample. No cross-reactivity was observed with coronaviruses OC43, NL63, 229E, SARS-CoV, nor with 92 clinical specimens containing common human respiratory viruses. We recommend using upE for screening and ORF1b for confirmation. .