Aspergillus fumigatus (A. fumigatus) is a widespread opportunistic pathogen. The conidia produced by A. fumigatus are colonized in lungs after inhalation, leading to aspergilloma, allergic bronchopulmonary aspergillosis (ABPA) and invasive aspergillosis (IA). Currently, 1，3-β-D-glucan and galactomannan are used as serum immunological indexes for clinical diagnosis of A. fumigatus infection. Asp f3 and Asp f4 are two of the major antigens of A. fumigatus, and the corresponding circulating antibodies are present in the sera of infected persons. In this study, anti-Asp f3 and anti-Asp f4 rabbit sera were used to carry out antigen epitope scanning. Eight Asp f3 and six Asp f4 minimal motifs were identified. These identified minimal motifs can be used for constructing chimera antigens to improve the specificity and sensitivity of the diagnosis for A. fumigatus infection.