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Since its first emergence in 2012, cases of infection with Middle East respiratory syndrome coronavirus (MERS-CoV) have continued to occur. At the end of January 2020, 2519 laboratory confirmed cases with a case-fatality rate of 34.3% have been reported. Approximately 84% of human cases have been reported in the tropical region of Saudi Arabia. The emergence of MERS-CoV has highlighted need for a rapid and accurate assay to triage patients with a suspected infection in a timely manner because of the lack of an approved vaccine or an effective treatment for MERS-CoV to prevent and control potential outbreaks. In this study, we present two rapid and visual nucleic acid assays that target the MERS-CoV UpE and N genes as a panel that combines reverse transcription recombinase polymerase amplification with a closed vertical flow visualization strip (RT-RPA-VF). This test panel was designed to improve the diagnostic accuracy through dual-target screening after referencing laboratory testing guidance for MERS-CoV. The limit of detection was 1.2×101 copies/μl viral RNA for the UpE assay and 1.2 copies/μl viral RNA for the N assay, with almost consistent with the sensitivity of the RT-qPCR assays. The two assays exhibited no cross-reactivity with multiple CoVs, including the bat severe acute respiratory syndrome related coronavirus (SARSr-CoV), the bat coronavirus HKU4, and the human coronaviruses 229E, OC43, HKU1 and severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Furthermore, the panel does not require sophisticated equipment and provides rapid detection within 30 min. This panel displays good sensitivity and specificity and may be useful to rapidly detect MERS-CoV early during an outbreak and for disease surveillance. Author summary: Symptoms of MERS are atypical, including fever, cough, shortness of breath and occasionally pneumonia and gastrointestinal symptoms. As a result, it is difficult to distinguish from other respiratory pathogens based on the clinical symptoms. MERS-CoV is routinely diagnosed using real-time RT-PCR. Real-time RT-PCR has great advantages in sensitivity and specificity compared with other molecular techniques; nevertheless, it is only suitable for well-equipped or central laboratories due to the demand for sophisticated alternating temperature and fluorescent capture instruments. Here, we developed and evaluated the MERS-CoV RT-RPA-VF assay. Compared to the gold standard real-time RT-PCR, the former was less time consuming and had lower equipment requirements. Thus, this panel may be preferred to rapidly detect MERS-CoV in low-resource settings, even as a point-of-care approach for the timely prevention and control of a pandemic. [ABSTRACT FROM AUTHOR]