Unraveling Hemoglobin D’s Influence: A Comprehensive Analysis of Clinicopathological Parameters in Hemoglobin D Patients

This prospective cross-sectional study was conducted at the Department of Hematology at Armed Forces Institute of Pathology Rawalpindi, Pakistan, from July 2023 to February 2025 after approval from Ethical Review Committee of the institute. Individuals being investigated for hemoglobinopathies in whom Hemoglobin D was detected, were included in the study. After detailed history and examination, investigations were performed including Complete Blood Counts (on Sysmex XN-3000), Capillary Zone Electrophoresis (on Sebia Capillarys 2 Flex-Piercing), High Performance Liquid Chromatography (on Bio-Rad D10) for differentiating Hb D-Punjab and D-Iran. Molecular studies (using PCR) were performed on samples in which a co-existing β thalassemia mutation was suspected on hemoglobin electrophoresis. Data collected was analyzed on Jamovi v2.4. Over 18 months, 2,171 individuals were tested for hemoglobinopathies, and Hb D, after excluding concomitant iron deficiency anemia, was detected in 106. Among these, 76 were found to have Hb D trait, 3 with homozygous Hb D disease, and 27 with compound heterozygous conditions. The compound heterozygous group included 21 patients of Hb D/β-thalassemia, 4 patients of Hb S/D, and 2 patients of Hb D/E. Hb D-Punjab accounted for 71% of the Hb D patients, and Hb D-Iran for the remaining 29%. Linear regression analysis revealed that MCH and RBC count showed significant positive correlations with Hb D levels. Molecular analysis identified specific β-thalassemia mutations in the Hb D/β-thalassemia cases, with IVS1-5 and FR 8-9 being the most common.