title: Clinical validation of 3 commercial real-time reverse transcriptase polymerase chain reaction assays for the detection of Middle East respiratory syndrome coronavirus from upper respiratory tract specimens.
abstract: Since discovery of Middle East respiratory syndrome coronavirus (MERS-CoV), a novel betacoronavirus first isolated and characterized in 2012, MERS-CoV real-time reverse transcriptase polymerase chain reaction (rRT-PCR) assays represent one of the most rapidly expanding commercial tests. However, in the absence of extensive evaluations of these assays on positive clinical material of different sources, evaluating their diagnostic effectiveness remains challenging. We describe the diagnostic performance evaluation of 3 common commercial MERS-CoV rRT-PCR assays on a large panel (n = 234) of upper respiratory tract specimens collected during an outbreak episode in Saudi Arabia. Assays were compared to the RealStar® MERS-CoV RT-PCR (Alton Diagnostics, Hamburg, Germany) assay as the gold standard. Results showed i) the TIB MolBiol® LightMix UpE and Orf1a assays (TIB MolBiol, Berlin, Germany) to be the most sensitive, followed by ii) the Anyplex™ Seegene MERS-CoV assay (Seegene, Seoul, Korea), and finally iii) the PrimerDesign™ Genesig® HCoV_2012 assay (PrimerDesign, England, United Kingdom). We also evaluate a modified protocol for the PrimerDesign™ Genesig® HCoV_2012 assay.
authors.affiliation: Pathology/Microbiology Division, College of Medicine, King Saud University, Riyadh, Saudi Arabia. Electronic address: dmohamed@ksu.edu.sa.
authors.forename: Deqa H
authors.initials: DH
authors.lastname: Mohamed
authors.affiliation: Pathology/Microbiology Division, College of Medicine, King Saud University, Riyadh, Saudi Arabia.
authors.forename: AbdulKarim F
authors.initials: AF
authors.lastname: AlHetheel
authors.affiliation: Transfusion Microbiology Department, College of Medicine, King Saud University, Riyadh, Saudi Arabia.
authors.forename: Hanat S
authors.initials: HS
authors.lastname: Mohamud
authors.affiliation: Central Blood Bank, Riyadh, Saudi Arabia.
authors.forename: Kamel
authors.initials: K
authors.lastname: Aldosari
authors.affiliation: Department of Pediatrics, College of Medicine, King Saud University, Riyadh, Saudi Arabia.
authors.forename: Fahad A
authors.initials: FA
authors.lastname: Alzamil
authors.affiliation: Pathology/Microbiology Division, College of Medicine, King Saud University, Riyadh, Saudi Arabia.
authors.forename: Ali M
authors.initials: AM
authors.lastname: Somily
mesh_terms: D018352:Coronavirus Infections / Q000382:microbiology*; D004196:Disease Outbreaks; D004739:England; D005858:Germany; D006801:Humans; D065207:Middle East Respiratory Syndrome Coronavirus / Q000235:genetics*; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D056910:Republic of Korea; D012137:Respiratory System / Q000382:microbiology*; D012141:Respiratory Tract Infections / Q000382:microbiology*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012529:Saudi Arabia
keywords: MERS-CoV; Orf1a; RT-PCR; Saudi Arabia; UpE