Gene amplification and detection of the TBEV by TBEV-specific RT-LAMP assay. (A) Real-time kinetics of the RT-LAMP amplification was monitored by real-time turbidimeter. In vitro transcribed RNA of TBEV Oshima strain was serially diluted to make 10-1 to 104 copies. The reaction was repeated three times and a representative result is shown. (B) Agarose gel electrophoresis profile of the RT-LAMP products. M indicates 100-bp DNA marker (Sigma). N indicates a sample containing no viral RNA. (C and D) Visual detection of the RT-LAMP products amplified in a reaction tube. Positive amplification is indicated by change of color to cloudy yellow (C) or by fluorescent green under UV irradiation (D). (E) Real-time kinetics of TaqMan rRT-PCR with TBEV-specific primer/probe set. In vitro transcribed RNA of Oshima was serially diluted to make 10-1 to 106 copies. The reaction was repeated three times and a representative result is shown.