Objective：This study aims to establish an immunological method for detecting specific antibody to Aspergillus fumigatus thioredoxin reductase（TR）in human sera，and evaluate its value in clinical prospect. Methods：The double-antigen sandwich ELISA for detecting TR antibody was applied with recombinant TR as coating and enzyme-labelled antigen. Reaction conditions were optimized by chessboard titration，and the sensitivity，specificity and repeatability for the ELISA were determined. Sera from 273 inpatients（50 cases with invasive aspergillosis，46 cases with invasive candidiasis，82 cases with Candida colonization and 95 cases with bacterial infection）and 200 healthy volunteers were collected and tested by the sandwich ELISA. The experiments results were compared with the previous results of indirect ELISA. Results：The TR was labeled with horseradish peroxidase（HRP）by the reformative sodium periodate method and the optimized condition were as follows：the coating concentration of TR was 1.0 μg/mL，the dilution of enzyme-labelled antigen was 1∶500，the blocking buffer was PBS-T containing 50 g/L dried skimmed milk，test serum was diluted to 1∶100. The intra- and inter-assay coefficients of variation for the developed ELISA were 11.2%，11.8%，12.3% and 10.9%，12.1%，13.5%，respectively. The blocking rate of sera with recombinant antigen was 94.4%. The absorbance value of 0.126 was chosen as cut-off value according to the ROC curve. The sensitivity of the sandwich ELISA for diagnosis of invasive aspergillosis was 80.0%（40/50），higher than that of indirect ELISA（83.3% vs. 72.2%） and the specificity was 95.5%（404/423） in this condition. Conclusion：A double-antigen sandwich ELISA for detecting the specific antibody to Aspergillus fumigatus TR was established successfully，which can be potentially applied in clinical diagnosis setting of invasive aspergillosis.