title: Establishment of a novel one-step reverse transcription loop-mediated isothermal amplification assay for rapid identification of RNA from the severe fever with thrombocytopenia syndrome virus.
abstract: As an emerging infectious disease, severe fever with thrombocytopenia syndrome virus (SFTSV) infection has been found in many areas of China. Suitable laboratory diagnostic method is urgently needed in clinical detections and epidemiological investigations. In this study, a modified, low-cost and rapid visualized one-step reverse transcription loop-mediated isothermal amplification (RT-LAMP) method for the detection of RNA from the SFTSV has been established. In order to avoid the risk of aerosol contamination and facilitate the naked eye to observe, a microcrystalline wax-dye capsule wrapping the highly sensitive DNA fluorescence dye SYBR Green I was added to the RT-LAMP reaction tube before the initiation of the assay. The detection limit of the established RT-LAMP assay was 10 fg template RNA per reaction mixture. The RT-LAMP assay was confirmed to be high specific to SFTSV, and no cross-reaction was found with the detection of the Chikungunya fever virus, Hemorrhagic Fever with Renal Syndrome virus (HFRSV), and Dengue fever virus. The assay was then applied for the detection of SFTSV RNA in 32 clinical serum samples and showed 94.4% consistence with the detection results of the real-time RT-PCR. The whole process, from sample preparation to result reporting, can be completed within 2h. This adapted, cost efficient and quick visualized RT-LAMP method is feasible for SFTSV field diagnosis in resource-limited field settings.
authors.affiliation: Sir Run Run Shaw Hospital, Zhejiang University School of Medicine, Hangzhou, Zhejiang 310016, China.
authors.forename: Haihong
authors.initials: H
authors.lastname: Xu
authors.forename: Lei
authors.initials: L
authors.lastname: Zhang
authors.forename: Guangqiang
authors.initials: G
authors.lastname: Shen
authors.forename: Cen
authors.initials: C
authors.lastname: Feng
authors.forename: Xinying
authors.initials: X
authors.lastname: Wang
authors.forename: Jie
authors.initials: J
authors.lastname: Yan
authors.forename: Yanjun
authors.initials: Y
authors.lastname: Zhang
mesh_terms: D002681:China; D003365:Costs and Cost Analysis; D025202:Molecular Diagnostic Techniques / Q000191:economics / Q000379:methods*; D021141:Nucleic Acid Amplification Techniques / Q000191:economics / Q000379:methods*; D010217:Phlebotomus Fever / Q000175:diagnosis* / Q000821:virology; D016856:Phlebovirus / Q000235:genetics / Q000302:isolation & purification*; D012367:RNA, Viral / Q000235:genetics / Q000302:isolation & purification*; D048348:Reverse Transcription; D012680:Sensitivity and Specificity; D013997:Time Factors
chemical_list: D012367:RNA, Viral
keywords: Detection; One-step; RT-LAMP; SFTSV