title: Performance and clinical validation of the RealStar MERS-CoV Kit for detection of Middle East respiratory syndrome coronavirus RNA.
abstract: BACKGROUND
  A highly pathogenic human coronavirus causing respiratory disease emerged in the Middle East region in 2012. In-house molecular diagnostic methods for this virus termed Middle East respiratory syndrome coronavirus (MERS-CoV) allowed sensitive MERS-CoV RNA detection in patient samples. Fast diagnosis is important to manage human cases and trace possible contacts.
  
  
  OBJECTIVE
  The aim of this study was to improve the availability of existing nucleic acid amplification-based diagnostic methods for MERS-CoV infections by providing a real-time RT-PCR kit, including an internal control and two target regions recommended by the World Health Organization (WHO). And to validate this kit (RealStar MERS-CoV RT-PCR kit 1.0, Altona Diagnostics GmbH, Hamburg, Germany) using clinical samples of one MERS-CoV case from Munich and respiratory samples of patients with other respiratory diseases.
  
  
  METHODS
  An internal amplification control was included into the RT-PCR assays targeting the genomic region upstream of the Envelope gene (upE) and within open reading frame (ORF) 1A. Based on these assays, a ready-to-use real-time RT-PCR kit featuring both the upE and ORF1A assays was developed, validated and compared to the established in-house versions.
  
  
  RESULTS
  The performance of both RT-PCR assays included in the kit is comparable to the in-house assays. They show high analytical sensitivity (upE: 5.3 copies/reaction; ORF1A: 9.3 copies/reaction), no cross-reactivity with other respiratory pathogens and detected MERS-CoV RNA in patient samples in almost the same manner as the in-house versions.
  
  
  CONCLUSIONS
  The kit is a valuable tool for assisting in the rapid diagnosis, patient management and epidemiology of suspected MERS-CoV cases.
authors.affiliation: Institute of Virology, University of Bonn Medical Centre, 53127 Bonn, Germany. Electronic address: corman@virology-bonn.de.
authors.forename: Victor Max
authors.initials: VM
authors.lastname: Corman
authors.affiliation: Altona Diagnostics GmbH, Mörkenstrasse 12, 22767 Hamburg, Germany.
authors.forename: Stephan
authors.initials: S
authors.lastname: Ölschläger
authors.affiliation: Klinikum Schwabing, Munich, Germany.
authors.forename: Clemens-Martin
authors.initials: CM
authors.lastname: Wendtner
authors.affiliation: Institute of Virology, University of Bonn Medical Centre, 53127 Bonn, Germany.
authors.forename: Jan Felix
authors.initials: JF
authors.lastname: Drexler
authors.affiliation: Altona Diagnostics GmbH, Mörkenstrasse 12, 22767 Hamburg, Germany.
authors.forename: Markus
authors.initials: M
authors.lastname: Hess
authors.affiliation: Institute of Virology, University of Bonn Medical Centre, 53127 Bonn, Germany.
authors.forename: Christian
authors.initials: C
authors.lastname: Drosten
mesh_terms: D018352:Coronavirus Infections / Q000175:diagnosis*; D003429:Cross Reactions; D006801:Humans; D065207:Middle East Respiratory Syndrome Coronavirus / Q000235:genetics / Q000302:isolation & purification*; D025202:Molecular Diagnostic Techniques / Q000379:methods* / Q000592:standards; D012367:RNA, Viral / Q000235:genetics / Q000302:isolation & purification*; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods* / Q000592:standards; D012015:Reference Standards; D012680:Sensitivity and Specificity
chemical_list: D012367:RNA, Viral
keywords: Coronavirus; Diagnostic assay; MERS-CoV; Middle East respiratory syndrome; PCR
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