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KMID : 0545120180280111928
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Journal of Microbiology and Biotechnology
2018 Volume.28 No. 11 p.1928 ~ p.1936
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Simple, Rapid and Sensitive Portable Molecular Diagnosis of SFTS Virus Using Reverse Transcriptional Loop-Mediated Isothermal Amplification (RT-LAMP)
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Baek Yun-Hee
Cheon Hyo-Soon
Park Su-Jin
Lloren Khristine Kaith S.
Ahn Su-Jeong
Jeong Ju-Hwan
Choi Won-Suk
Yu Min-Ah
Kwon Hyeok-Il
Kwon Jin-Jung
Kim Eun-Ha
Kim Young-Il
Antigua Khristine Joy C.
Kim Seok-Yong
Jeong Hye-Won
Choi Young-Ki
Song Min-Suk
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Abstract
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Recently, human infections caused by severe fever with thrombocytopenia syndrome virus (SFTSV), which can lead to fatality, have dramatically increased in East Asia. With the unavailability of vaccines or antiviral drugs to prevent and/or treat SFTSV infection, early rapid diagnosis is critical for prevention and control of the disease. Here, we report the development of a simple, rapid and sensitive portable detection method for SFTSV infection applying reverse transcription?loop mediated isothermal amplification (RT-LAMP) combined with one-pot colorimetric visualization and electro-free reaction platform. This method utilizes a pocket warmer to facilitate diagnosis in a resource-limited setting. Specific primers were designed to target the highly-conserved region of L gene of SFTSV. The detection limit of the RT-LAMP assay was approximately 100 viral genome copies from three different SFTSV strains. This assay exhibited comparable sensitivity to qRT-PCR and 10-fold more sensitivity than conventional RT-PCR, with a rapid detection time of 30 to 60 minutes. The RT-LAMP assay using SFTSV clinical specimens has demonstrated a similar detection rate to qRT-PCR and a higher detection rate compared to conventional RT-PCR. Moreover, there was no observed cross-reactive amplification of other human infectious viruses including Japanese Encephalitis Virus (JEV), Dengue, Enterovirus, Zika, Influenza and Middle East Respiratory Syndrome Coronavirus (MERS-CoV). This highly sensitive, electro- and equipment-free rapid colorimetric visualization method is feasible for resource-limited SFTSV field diagnosis.
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KEYWORD
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SFTSV, RT-LAMP, colorimetric visualization, pocket warmer
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