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
Autor(es): Xu Haihong, Zhang Lei, Shen Guangqiang, Feng Cen, Wang Xinying, Yan Jie, Zhang Yanjun
Resumo: 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 - epidemiological investigations. In this study, a modified, low-cost - 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 - 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, - no cross-reaction was found with the detection of the Chikungunya fever virus, Hemorrhagic Fever with Renal Syndrome virus (HFRSV), - Dengue fever virus. The assay was then applied for the detection of SFTSV RNA in 32 clinical serum samples - 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 - quick visualized RT-LAMP method is feasible for SFTSV field diagnosis in resource-limited field settings.
Palavras-Chave: Detection; One-step; RT-LAMP; SFTSV
Imprenta: Journal of Virological Methods, v. 194, n. 1-2, p. 21-25, 2013
Identificador do objeto digital: 10.1016/j.jviromet.2013.07.037
Descritores: Chikungunya virus - DNA ; Chikungunya virus - Pathogenesis ; Chikungunya virus - RNA ; Chikungunya virus - Viral infections ; Chikungunya virus - Molecular methods ; Chikungunya virus - PCR detection ; Chikungunya virus - Real Time PCR ; Chikungunya virus - RT-PCR ; Chikungunya Virus - Virus ; Chikungunya virus - Molecular screening ; Chikungunya virus - Chikungunya fever ; Chikungunya virus - Dengue ; Chikungunya virus - Public health
Data de publicação: 2013
