Mycotoxin contamination in edible oil has been a serious problem in recent years. The feasibility of ATR- FTIR spectroscopy with Principal Component Analysis (PCA) was investigated for detecting 15-AcDON in crude and refined corn oils. Corn oils contaminated artificially with 15-acetyldeoxynivalenol (15-AcDON) and Fusarium asiaticum (defined as “Fusarium-contaminated corn oil”) and only with 15-AcDON (defined as “Fusarium-free corn oil”) were tested to simulate 15-AcDON contamination in crude oil and in refined oil, respectively. The contaminations with 15-AcDON at different levels were discriminated by a PCA score plot for both contaminated corn oils. The specific IR bands of 15-AcDON were revealed for each oil, which could be assigned to the symmetric stretching vibration of the ester (O)CeO. By comparison between the spectra of both oils, no interference of the F. asiaticum in the detection of 15-AcDON was found, however, the difference of the specific band region for 15-AcDON between both oils was observed. The results suggested that ATR-FTIR spectroscopy could be a promising method for rapid screening of 15- AcDON in crude and refined corn oils.