Real-Time Reverse Transcription-PCR Assay Panel for Middle East Respiratory Syndrome Coronavirus |
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| Authors: | Xiaoyan Lu Brett Whitaker Senthil Kumar K. Sakthivel Shifaq Kamili Laura E. Rose Luis Lowe Emad Mohareb Emad M. Elassal Tarek Al-sanouri Aktham Haddadin Dean D. Erdman |
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| Affiliation: | aDivision of Viral Diseases, Centers for Disease Control and Prevention, Atlanta, Georgia, USA;bDivision of Preparedness and Emerging Infections, Centers for Disease Control and Prevention, Atlanta, Georgia, USA;cU.S. Naval Medical Research Unit 3, Cairo, Egypt;dCentral Public Health Laboratory, Ministry of Health, Amman, Jordan |
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| 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. |
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