African swine fever（ASF）is a transboundary swine disease，causing huge economic losses for swine industry in the world. At present，the prevalence of the disease has been spreading widely. In order to prevent the disease from being introduced into China，it is urgent to establish a fast，simple and reliable method. In this study，the specific primers and exo probes were designed and synthesized by analyzing the conserved region of B646L gene of African swine fever virus（ASFV）. A real-time fluorescence RPA method for detection of ASFV was established. The minimal detection limit of the novel real-time RPA assay was 16 copies per reaction，and the detection could be finished in 20 minutes. Cross reactivity with other swine associated viruses was not observed. The sensitivity and specificity of the assay was comparable to that of the real-time PCR. 100 domestic samples were detected by this method，and the results were negative. The real-time RPA assay developed in this study was of great significance of simple，fast，high sensitivity and strong specificity，with wide application value in clinical differential diagnosis，laboratory quarantine and pathogen surveillance.