<p>Alpha-thalassemia reported to be a genetic modifier in β-thalassemia characterized with hematological and clinical heterogeneity. This study was carried out to analyze the effect of α⁺-thalassemia (–α^3.7 and –α^4.2) on the hematological profile of β-thalassemia trait cases with IVS1-5 (G→C) mutation.&#xa0;In this cross-sectional study, cases advised to undergo screening hemoglobinopathies were investigated for complete blood count and hemoglobin fractions. Individuals with suspicion of β-thalassemia trait were subjected to analysis for β-thalassemia mutations and α+-thalassemia (–α^3.7 and –α^4.2). Cases with β-thalassemia trait carrying only the IVS1-5 (G→C) mutation were further analyzed for the possible effect of α⁺-thalassemia on the hematological profile using Kruskal-Wallis test.&#xa0;A total of 118 β-thalassemia trait cases with IVS 1–5 (G→C) mutation were included, comprising 20 (16.9%) pediatric and 64 (54.20%) female cases. Co-inherited α⁺-thalassemia (–α^3.7 and –α^4.2) was detected in 48 (40.68%) cases, including 33 cases in heterozygous and 15 cases in homozygous state. Hematological parameters such as MCV, MCH, Ret-He, macro-R, and hyper-He were highest in homozygous α⁺-thalassemia, followed by heterozygous and normal α-globin genotypes. Conversely, IRF, Micro-R, and Hypo-He showed an inverse pattern. Similar trends were observed across gender and adult age groups.&#xa0;Co-inheritance of α+-thalassemia in individuals with β-thalassemia trait is associated with a more favorable hematological profile. Beyond conventional red cell indices such as MCV and MCH, incorporation of advanced parameters including micro-R, macro-R, hypo-He, hyper-He, and IRF may provide a more comprehensive diagnostic approach to these genetic disorders.</p>