Background: Molecular detection of Middle East respiratory syndrome coronavirus (MERS-CoV) using real-time reverse transcription (rRT)-PCR assays is the method of choice for diagnosis of MERS. We evaluated the performance of the Power. Chek MERS (upE & ORF1a) real-time PCR Kit (Power. Chek MERS assay; Kogene Biotech, Korea) a one-step rRT-PCR assay for the qualitative detection of MERS-CoV. Methods: We evaluated Power. Chek MERS assay performance in comparison with nested RT-PCR and sequencing of the RNA-dependent RNA polymerase (Rd. Rp) and N genes. To evaluate diagnostic sensitivity and specificity, 100 clinical specimens (50 positive and 50 negative for MERS-CoV) were simultaneously tested by using the Power. Chek MERS and sequencing assays. Assay performance, including limit of detection and precision, was evaluated in vitro by using MERS-CoV RNA transcripts. Analytical specificity was evaluated with a diverse collection of 16 respiratory virus-positive clinical specimens and 14 respiratory bacterial isolates. Results: The 95% limits of detection of the Power. Chek MERS assay for the upE and the open rading frame (ORF)1a were 16.2 copies/ฮผL and 8.2 copies/ฮผL, respectively. No cross-reactivity was observed. The diagnostic sensitivity and specificity of the Power. Chek MERS assay were both 100% (95% confidence interval, 91.1-100%). Conclusions: The Power. Chek MERS assay is a straightforward and accurate assay for detecting MERS-CoV RNA. The assay will be a useful tool for the rapid diagnosis of MERS and could prove especially important for MERS outbreak control.