: We aimed to evaluate the seminal biochemical markers and serum fertility hormones in men with or without infertility in province of Basrah-Iraq. From 113 men volunteers, 57 were with primary and 56 had secondary infertility, while 50 fertile men were taken as controls. Their fasting BMI, serum, LH, FSH, PRL, Ts, seminal fluid resistin, NAG, and total protein were determined by ELISA methods. Seminal fluid citric Acid, fructose and ACP were measured by standard colorimetric methods. Compared with normal controls, the results indicated that patient’s men with 1o IM and 2o IM had significantly higher levels of FSH (p<0.01), LH (p<0.05), PRL (p<0.01), resistin (p<0.05), ACP (p<0.01), and fructose (p<0.01), and significantly decreases in Ts (p<0.01), NAG (p<0.01) and CA (p<0.01) and total protein (p<0.05), while BMI levels were not significantly different (p>0.05). In conclusion, the findings further demonstrated that determination of serum reproductive hormones and seminal fluid biochemical parameters may give a useful information for clinical studies in the status of men infertility as well as can be used as a biomarker for detection of early-stage of infertility and treatment success of men fertility disorders.