The present study aimed to evaluate the prevalence of alpha-thalassemia and glucose-6-phosphate dehydrogenase (G6PD) deficiency in sickle cell trait (SCT) patients to determine its effect on red blood cells (RBC) parameters. This cross-sectional study was conducted on 102 blood samples obtained from children and teenagers with SCT aged between 5 and 18 years old who were referred to Shahid Beqaei Hematology and Oncology Hospital and Abuzar Children's Hospital in Ahvaz city (Iran) from October 2021 to November 2022. About 5 mL of blood was collected via venipuncture from each patient and used to run G6PD, complete blood count, and hemoglobin (Hb) electrophoresis tests. The data were analyzed using SPSS version 22, and the significance level in all tests was considered less than 0.05. Results showed that the prevalence of heterozygous and homozygous alpha-thalassemia and iron deficiency anemia (IDA) in the examined sample was 18.63%, 18.63%, and 10.78%, respectively. Also, 13.72% of patients suffered from G6PD deficiency. The results imply that G6PD deficiency may increase the severity of anemia in SCT patients. Therefore, it is necessary to screen all SCT patients for G6PD deficiency to ensure that their condition is not exacerbated during unexpected events such as diseases or stress.