The invention discloses a fluorescent quantitative PCR (Polymerase Chain Reaction) detection reagent, a kit and a detection method for an African swine fever virus (ASFV). The detection reagent is targeted to a conserved segment of an ASFV VP72 gene, and comprises a pair of specific primers, a specific probe and an internal labeling probe. The kit comprises a PCR mixed solution, Taq DNA polymerase, DEPC (diethylpyrocarbonate)-H2O, an ASFV positive quality product, an ASFV negative quality product, a quantification standard substance and a pseudovirus internal labeling solution, wherein the PCR mixed solution comprises a pair of specific primers, a specific probe and an internal labeling probe. The detection method comprises the steps of extraction of total DNAs, preparation of reaction components, making of a standard curve by diluting the standard substance, amplifying and result judgment. The reagent and the kit have the advantages of high specificity, high sensitivity, no pollution and high flux, and can be used for accurately detecting infection, inapparent infection or continuous carrying of a toxic host of low-content ASFV. The detection method is quick, specific and sensitive, and can be used for quantitatively detecting a large quantity of samples at the same time.