Occult hepatitis B (OHB) is characterized by the presence of hepatitis B virus (HBV) DNA in the blood of individuals who test negative for the hepatitis B surface antigen (HBsAg). OHB in blood donors can lead to HBV transmission through transfusions, yet the prevalence of OHB in Basrah, Iraq, is unknown. This study aimed to determine the prevalence of OHB in blood donation centers in Basrah and investigate the immune response to HBV in OHB-positive donors. We recruited 450 blood donors and categorized them into four groups based on HBV markers: the HBsAg-negative/HBsAb-negative/HBcAb-positive group, the recovery group (HBsAg-negative/HBsAb-positive/HBcAb-positive), the patient group (HBsAg-positive/HBsAb-negative/HBcAb-positive), and the apparently healthy group (negative for all HBV markers). We measured levels of IgG, IgM, complement components (C3 and C4), ALT, AST, and serum ALP in OHB-positive donors. Of the 450 donors, 97 (21.6%) were OHB-positive. IgG levels were significantly higher than IgM levels in OHB-positive donors. Healthy and HBsAg-negative/HBsAb-positive donors had significantly lower C3 levels than patients. IgG levels were significantly higher than IgM in both the patient and recovery groups. C3 levels were higher than C4 levels in all groups. The serum ALP level was significantly higher in the patient group. OHB prevalence in Basrah blood donors is high, indicating the potential for HBV transmission. OHB-positive donors showed an immune response to HBV. Our study provides insights into OHB prevalence and immune response in Basrah, with implications for diagnostic and therapeutic approaches in blood donation centers.