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Open Access 01-12-2014 | Research

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

Authors: Kazuya Shirato, Takuya Yano, Syouhei Senba, Shigehiro Akachi, Takashi Kobayashi, Takamichi Nishinaka, Tsugunori Notomi, Shutoku Matsuyama

Published in: Virology Journal | Issue 1/2014

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.

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Title: Detection of Middle East respiratory syndrome coronavirus using reverse transcription loop-mediated isothermal amplification (RT-LAMP)
Authors: Kazuya Shirato
Takuya Yano
Syouhei Senba
Shigehiro Akachi
Takashi Kobayashi
Takamichi Nishinaka
Tsugunori Notomi
Shutoku Matsuyama
Publication date: 01-12-2014
Publisher: BioMed Central
Published in: Virology Journal / Issue 1/2014
Electronic ISSN: 1743-422X
DOI: https://doi.org/10.1186/1743-422X-11-139

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

Abstract

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Methods

Results

Conclusions

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