β-thalassemia (BT) is one of the most frequent hereditary hematologic diseases. The major complication of BT patients is iron overload that could lead to many organs damage and elevated mortality. The purpose of this study was to evaluate the role of serum amyloid A (SAA) in differentiate BT patients from other types of anemia and healthy controls. It was also to evaluate its association with iron overload and ineffective erythropoiesis in BT. Elevated SAA was related (P<0.0001) to BT (33.3 (31.2-48.5) ng/mL) compared to patients with other types of anaemia and healthy controls (14.7 (10.9-20.2) and 3.0 (1.0-6.5) ng/mL, respectively). It had a great power (AUC=1.00, P<0.0001) to discriminate BT patients. Its levels was increased (P=0.0399) in patients with ferritin>1000 ng/mL compared to those with <1000 ng/mL. It had a good ability to differentiate cases with iron overload (AUC=0.85, P=0.0129). Elevated SAA levels were significantly (P<0.05) correlated to elevated levels of ferritin (r=0.768), bilirubin (r=0.438) and ALT (r=0.675) and AST (r=0.678) activities, reduced hemoglobin (r=-0.691) and ineffective erythropoiesis [RBCs count (r=-0.256), MCV (r= 0.397) and PCV (r=-0.571)]. In conclusion, high level of SAA as new biomarker in patients with major BT is related to iron overload, mild/severe anemia and ineffective erythropoiesis.