title: Real-time reverse transcription-PCR assay panel for Middle East respiratory syndrome coronavirus.
abstract: A new human coronavirus (CoV), subsequently named Middle East respiratory syndrome (MERS)-CoV, was first reported in Saudi Arabia in September 2012. In response, we developed two real-time reverse transcription-PCR (rRT-PCR) assays targeting the MERS-CoV nucleocapsid (N) gene and evaluated these assays as a panel with a previously published assay targeting the region upstream of the MERS-CoV envelope gene (upE) for the detection and confirmation of MERS-CoV infection. All assays detected ≤10 copies/reaction of quantified RNA transcripts, with a linear dynamic range of 8 log units and 1.3 × 10(-3) 50% tissue culture infective doses (TCID50)/ml of cultured MERS-CoV per reaction. All assays performed comparably with respiratory, serum, and stool specimens spiked with cultured virus. No false-positive amplifications were obtained with other human coronaviruses or common respiratory viral pathogens or with 336 diverse clinical specimens from non-MERS-CoV cases; specimens from two confirmed MERS-CoV cases were positive with all assay signatures. In June 2012, the U.S. Food and Drug Administration authorized emergency use of the rRT-PCR assay panel as an in vitro diagnostic test for MERS-CoV. A kit consisting of the three assay signatures and a positive control was assembled and distributed to public health laboratories in the United States and internationally to support MERS-CoV surveillance and public health responses.
authors.affiliation: Division of Viral Diseases, Centers for Disease Control and Prevention, Atlanta, Georgia, USA.
authors.forename: Xiaoyan
authors.initials: X
authors.lastname: Lu
authors.forename: Brett
authors.initials: B
authors.lastname: Whitaker
authors.forename: Senthil Kumar K
authors.initials: SK
authors.lastname: Sakthivel
authors.forename: Shifaq
authors.initials: S
authors.lastname: Kamili
authors.forename: Laura E
authors.initials: LE
authors.lastname: Rose
authors.forename: Luis
authors.initials: L
authors.lastname: Lowe
authors.forename: Emad
authors.initials: E
authors.lastname: Mohareb
authors.forename: Emad M
authors.initials: EM
authors.lastname: Elassal
authors.forename: Tarek
authors.initials: T
authors.lastname: Al-sanouri
authors.forename: Aktham
authors.initials: A
authors.lastname: Haddadin
authors.forename: Dean D
authors.initials: DD
authors.lastname: Erdman
mesh_terms: D000328:Adult; D017934:Coronavirus / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification*; D018352:Coronavirus Infections / Q000175:diagnosis* / Q000821:virology; D006801:Humans; D007223:Infant; D007231:Infant, Newborn; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D011933:Reagent Kits, Diagnostic; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D012141:Respiratory Tract Infections / Q000175:diagnosis* / Q000821:virology; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D014481:United States; D014486:United States Food and Drug Administration
chemical_list: D011933:Reagent Kits, Diagnostic