Background The first documented case of Middle East Respiratory Syndrome coronavirus (MERS-CoV) occurred in 2012, and outbreaks have continued ever since, mainly in Saudi Arabia. MERS-CoV is primarily diagnosed using a real-time RT-PCR assay, with at least two different genomic targets required for a positive diagnosis according to the case definition of The World Health Organization (WHO) as of 3 July 2013. Therefore, it is urgently necessary to develop as many specific genetic diagnostic methods as possible to allow stable diagnosis of MERS-CoV infections. Methods Reverse transcription-loop-mediated isothermal amplification (RT-LAMP) is a genetic diagnostic method used widely for the detection of viral pathogens, which requires only a single temperature for amplification, and can be completed in less than 1 h. This study developed a novel RT-LAMP assay for detecting MERS-CoV using primer sets targeting a conserved nucleocapsid protein region. Results The RT-LAMP assay was capable of detecting as few as 3.4 copies of MERS-CoV RNA, and was highly specific, with no cross-reaction to other respiratory viruses. Pilot experiments to detect MERS-CoV from medium containing pharyngeal swabs inoculated with pre-titrated viruses were also performed. The RT-LAMP assay exhibited sensitivity similar to that of MERS-CoV real-time RT-PCR. Conclusions These results suggest that the RT-LAMP assay described here is a useful tool for the diagnosis and epidemiologic surveillance of human MERS-CoV infections.
【저자키워드】 RT-LAMP, MERS coronavirus (MERS-CoV), Meddle East respiratory syndrome (MERS), Genetic diagnostic method, 【초록키워드】 coronavirus, Genetic, Diagnosis, virus, MERS-CoV, Saudi Arabia, nucleocapsid protein, amplification, sensitivity, infections, outbreak, target, WHO, temperature, isothermal amplification, respiratory, real-time RT-PCR, genomic, Diagnostic method, Epidemiologic surveillance, Middle East, World Health Organization, Pharyngeal swab, medium, positive, viral pathogens, Result, described, performed, detect, develop, occurred, conserved, primer set, inoculated, diagnosed, required, exhibited, less, MERS-CoV RNA, Pilot experiment, 【제목키워드】 detection, reverse transcription, isothermal amplification, Middle East, respiratory syndrome coronavirus,