Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Simple, Rapid and Sensitive Portable Molecular Diagnosis of SFTS Virus Using Reverse Transcriptional Loop-Mediated Isothermal Amplification (RT-LAMP)

  • Baek, Yun Hee (Department of Microbiology, Chungbuk National University College of Medicine and Medical Research Institute) ;
  • Cheon, Hyo-Soon (Department of Microbiology, Chungbuk National University College of Medicine and Medical Research Institute) ;
  • Park, Su-Jin (Department of Microbiology, Chungbuk National University College of Medicine and Medical Research Institute) ;
  • Lloren, Khristine Kaith S. (Department of Microbiology, Chungbuk National University College of Medicine and Medical Research Institute) ;
  • Ahn, Su Jeong (Department of Microbiology, Chungbuk National University College of Medicine and Medical Research Institute) ;
  • Jeong, Ju Hwan (Department of Microbiology, Chungbuk National University College of Medicine and Medical Research Institute) ;
  • Choi, Won-Suk (Department of Microbiology, Chungbuk National University College of Medicine and Medical Research Institute) ;
  • Yu, Min-Ah (Department of Microbiology, Chungbuk National University College of Medicine and Medical Research Institute) ;
  • Kwon, Hyeok-il (Department of Microbiology, Chungbuk National University College of Medicine and Medical Research Institute) ;
  • Kwon, Jin-Jung (Department of Microbiology, Chungbuk National University College of Medicine and Medical Research Institute) ;
  • Kim, Eun-Ha (Department of Microbiology, Chungbuk National University College of Medicine and Medical Research Institute) ;
  • Kim, Young-il (Department of Microbiology, Chungbuk National University College of Medicine and Medical Research Institute) ;
  • Antigua, Khristine Joy C. (Department of Microbiology, Chungbuk National University College of Medicine and Medical Research Institute) ;
  • Kim, Seok-Yong (Department of Microbiology, Chungbuk National University College of Medicine and Medical Research Institute) ;
  • Jeong, Hye Won (Department of Internal Medicine, Chungbuk National University College of Medicine and Medical Research Institute) ;
  • Choi, Young Ki (Department of Microbiology, Chungbuk National University College of Medicine and Medical Research Institute) ;
  • Song, Min-Suk (Department of Microbiology, Chungbuk National University College of Medicine and Medical Research Institute)
  • Received : 2018.06.12
  • Accepted : 2018.09.12
  • Published : 2018.11.28
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Abstract

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 $10^0$ 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.

Keywords

References

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