The invention discloses a primer, a probe and a detection method for detecting nodular skin disease virus by RAA fluorescence method, wherein the primer and probe are suitable for RAA fluorescence method detection, and can accurately detect nodular skin disease virus plasmid , has no cross-reaction with mycoplasma, bovine infectious rhinotracheitis virus, bovine viral diarrhea virus, bovine parainfluenza virus, bovine respiratory syncytial virus, goat pox virus, and sheep pox virus, with a specificity of 100%; the detection method is rapid, It is easy to realize high-throughput, while reducing detection time and detection cost. The method for rapid detection of nodular skin disease virus DNA based on RAA fluorescence method provided by the present invention has high sensitivity, and the detection sensitivity of each reaction reaches 10copies / reaction.