title: Assays for laboratory confirmation of novel human coronavirus (hCoV-EMC) infections.
abstract: We present a rigorously validated and highly sensitive confirmatory real-time RT-PCR assay (1A assay) that can be used in combination with the previously reported upE assay. Two additional RT-PCR assays for sequencing are described, targeting the RdRp gene (RdRpSeq assay) and N gene (NSeq assay), where an insertion/deletion polymorphism might exist among different hCoV-EMC strains. Finally, a simplified and biologically safe protocol for detection of antibody response by immunofluorescence microscopy was developed using convalescent patient serum.
authors.affiliation: Institute of Virology, University of Bonn Medical Centre, Bonn, Germany.
authors.forename: V M
authors.initials: VM
authors.lastname: Corman
authors.forename: M A
authors.initials: MA
authors.lastname: Müller
authors.forename: U
authors.initials: U
authors.lastname: Costabel
authors.forename: J
authors.initials: J
authors.lastname: Timm
authors.forename: T
authors.initials: T
authors.lastname: Binger
authors.forename: B
authors.initials: B
authors.lastname: Meyer
authors.forename: P
authors.initials: P
authors.lastname: Kreher
authors.forename: E
authors.initials: E
authors.lastname: Lattwein
authors.forename: M
authors.initials: M
authors.lastname: Eschbach-Bludau
authors.forename: A
authors.initials: A
authors.lastname: Nitsche
authors.forename: T
authors.initials: T
authors.lastname: Bleicker
authors.forename: O
authors.initials: O
authors.lastname: Landt
authors.forename: B
authors.initials: B
authors.lastname: Schweiger
authors.forename: J F
authors.initials: JF
authors.lastname: Drexler
authors.forename: A D
authors.initials: AD
authors.lastname: Osterhaus
authors.forename: B L
authors.initials: BL
authors.lastname: Haagmans
authors.forename: U
authors.initials: U
authors.lastname: Dittmer
authors.forename: F
authors.initials: F
authors.lastname: Bonin
authors.forename: T
authors.initials: T
authors.lastname: Wolff
authors.forename: C
authors.initials: C
authors.lastname: Drosten
mesh_terms: D017934:Coronavirus / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D018352:Coronavirus Infections / Q000175:diagnosis* / Q000821:virology; D005455:Fluorescent Antibody Technique; D005858:Germany; D006801:Humans; D007753:Laboratories / Q000592:standards; D012150:Polymorphism, Restriction Fragment Length; D012367:RNA, Viral / Q000097:blood / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA; D014773:Virology / Q000379:methods
chemical_list: D012367:RNA, Viral