[Objective] Establish a simple and rapid molecular detection of Yellow fever virus. [Methods] The reserve transcription loop-mediated isothermal amplification (RT-LAMP) that amplifies RNA with high specificity and rapidity under an isothermal condition was applied for rapid detection of Yellow fever virus. A set of primers were designed specifically to identify six special areas of Yellow fever virus’s E gene. [Results] The results indicated that the method had high specificity, and no cross amplification occurred with other arbovirus RNAs. The sensitivity of the RT-LAMP assay was 0.066 pg, which was 100 times higher than that of RT-PCR. The detection results of simulation samples were consistent with expected. [Conclusion] The established rapid detection method for Yellow fever virus that can be carried out in a simple experimental environment has been proved to be specific and sensitive.