Context: Seminal fluid analysis (SFA) is the most important investigative tool for evaluation of male infertility. However, it is limited in detecting causes of sperm abnormalities while some individuals with normal SFA are still considered infertile. Evaluating sperm deoxyribonucleic acid (DNA) integrity as an ancillary investigative tool to SFA will enhance the investigation of infertility. Aims: The aim is to assess the prevalence and severity of sperm DNA damage based on semen level of 8-hydroxydeoxyguanosine (8-OHdG) in males with and without abnormal SFA. Settings and Design: This was a descriptive cross-sectional study of 120 males with at least one SFA parameter abnormalities as test controls and 120 normal controls. Subjects and Methods: Seminal 8-OHDG was assayed as a marker of sperm DNA damage using ELISA method. Statistical Analysis Used: Normally distributed data were expressed as mean ± standard deviation otherwise expressed as median and interquartile range. Results: The mean ages of subjects and controls were 35.84 ± 6.0 vs. 36.22 ± 7.56 years. The mean seminal 8-OHDG was significantly higher among subjects than among controls (15.21 ± 3.80 ng/ml vs. 12.45 ± 4.0 ng/ml, P = 0.015). The reference value of seminal 8-OHDG obtained in this study was 4.45–20.45 ng/ml and the prevalence of sperm DNA damage among subjects compared to controls was 10.8% versus 3.3%, P = 0.024. Severe DNA damage corresponding to sperm DNA fragmentation index of >30% was 3.3% in subjects and was not present in any male partner in the control group. Significant sperm DNA damage was also associated with reduced sperm count (P = 0.043), while its association with reduced sperm motility was not statistically significant in this study. Conclusions: The prevalence and severity of sperm DNA damage were more significant among males with, than those without, abnormal SFA parameters.