Detection of Middle East respiratory syndrome coronavirus using reverse transcription loop-mediated isothermal amplification (RT-LAMP)

Virology Journal

Shutoku Matsuyama²  Tsugunori Notomi¹  Takamichi Nishinaka³  Takashi Kobayashi³  Shigehiro Akachi³  Syouhei Senba¹  Takuya Yano³  Kazuya Shirato² 

[1] Eiken Chemical Co. Ltd., 4-19-9 Taito, Taito-ku, Tokyo 110-8408, Japan;Laboratory of Acute Respiratory Viral Diseases and Cytokines, Department of Virology III, National Institute of Infectious Disease, Laboratory of Acute Respiratory Viral Diseases and Cytokines, 4-7-1 Gakuen, Musashimurayama, Tokyo 208-0011, Japan;Mie Prefecture Health and Environment Research Institute, 3684-11 Sakura-cho, Yokkaichi, Mie 512-1211, Japan

关键词: Genetic diagnostic method;    RT-LAMP;    MERS coronavirus (MERS-CoV);    Meddle East respiratory syndrome (MERS);   

Others  :  1148762
DOI  :  10.1186/1743-422X-11-139

 received in 2014-04-25, accepted in 2014-08-04,  发布年份 2014

【 摘 要 】

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.

【 授权许可】

   
2014 Shirato et al.; licensee BioMed Central Ltd.

【 图 表 】

Figure 1.

Figure 2.

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