Email updates

Keep up to date with the latest news and content from Virology Journal and BioMed Central.

Open Access Highly Accessed Research

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

Kazuya Shirato1*, Takuya Yano2, Syouhei Senba3, Shigehiro Akachi2, Takashi Kobayashi2, Takamichi Nishinaka2, Tsugunori Notomi3 and Shutoku Matsuyama1

Author Affiliations

1 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

2 Mie Prefecture Health and Environment Research Institute, 3684-11 Sakura-cho, Yokkaichi, Mie 512-1211, Japan

3 Eiken Chemical Co. Ltd., 4-19-9 Taito, Taito-ku, Tokyo 110-8408, Japan

For all author emails, please log on.

Virology Journal 2014, 11:139  doi:10.1186/1743-422X-11-139

Published: 8 August 2014

Abstract

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.

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