objective:sperm dna fragmentation has been assessed as a positive predictor of fertility potential in recent years. the aim of this study is using the acridine orange test(aot) to analyze the relationship between sperm dna fragmentation indexes(dfi) and semen conventional parameters of infertility men. methods:semen samples were collected from 376 infertility male patients in our andrology outpatient department.the samples were analyzed by improved neubauer haemocytometer chamber and computer-aided sperm analysis(casa) system to obtain concentration and motility parameters. all the samples were stained by shorr staining method and the smears were analyzed according to tygerberg strict criteria recommended by world health organization(who) to obtain morphology parameters. the dfi as marker of sperm dna damage determined using the aot. results:significant negative correlations were noted between dfi and sperm concentration(r=-0.150,p=0.003),motility(r=-0.114,p=0.028)and morphology(r=-0.155,p=0.003). no significant correlations were found between dfi and sperm progressive motility (r=-0.089,p=0.085). conclusion:sperm assessment for dna status using ao is reliable and shows good correlation with sperm count,motility and morphology. assessment of sperm dna fragmentation with aot measures a new parameter of male fertility.