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.