Purpose: We performed and analyzed the DFI test and semen test based on the WHO criteria and compared the two using semen factors. Methods: DFI tests and general semen tests were conducted in accordance with the WHO criteria, and correlations between the two tests were investigated. The WHO criteria were set as the cutoff value for each of the following factors: fluid volume, concentration, motility, and normal morphology, and compared with the DFI results.Results: The subjects had a mean DFI of 15.6±11.5% and the DFI increased with age. In contrast, the concentration, motility, and normal morphology decreased as the DFI increased. Patients who satisfied the WHO criteria in terms of sperm concentration, motility, and normal morphology had a significantly lower DFI than the group that did not satisfy the criteria. Therefore, evaluation with a general semen test based on the WHO criteria should be regarded as a qualitative evaluation of all factors other than fluid volume.Conclusions: Male infertility due to sperm DNA fragmentation should be suspected when in vitro fertilization (IVF) results are poor despite normal semen findings based on the WHO criteria. The results of this study suggest that the DFI test may be an important evaluation in follow-up semen analyses.