Clinico-Hematological Profile of Double Heterozygous Hemoglobinopathies Diagnosed by High-Performance Liquid...

Background Hemoglobinopathies are inherited disorders of hemoglobin synthesis and structure that continue to impose a significant health burden, particularly in the Indian subcontinent, where genetic diversity, endogamy, and consanguinity contribute to their high prevalence. Double heterozygous hemoglobinopathies show considerable clinical and hematological variability, and limited data are available from tribal-predominant populations of eastern India. Our study was undertaken to assess the clinico-hematological profile of double heterozygous hemoglobinopathies and to evaluate the utility of high-performance liquid chromatography (HPLC) in their accurate detection and characterization. Methods This study was conducted in the Department of Pathology at a tertiary care teaching hospital in eastern India over 18 months. A total of 70 patients with confirmed double heterozygous hemoglobinopathies on HPLC were included. Hematological parameters were analyzed using an automated Sysmex hematology analyzer (Sysmex Corporation, Kobe, Hyogo, Japan), and definitive diagnosis was established using cation-exchange HPLC on the Bio-Rad VARIANT II Hemoglobin Testing System (Bio-Rad Laboratories, Inc., Hercules, California, United States). Statistical analysis was performed using IBM SPSS Statistics for Windows, version 27 (IBM Corp., Armonk, New York, United States), and comparisons between groups were made using the unpaired Student's t-test, Chi-square test, or Fisher's exact test as appropriate. Results Of the 70 patients included in the study, 60 (85.7%) had Sβ-thalassemia and 10 (14.3%) had HbE-β-thalassemia. Most patients belonged to the age group of 11-20 years, with a mean age of 15.1 ± 9.9 years. Pallor and splenomegaly were the most frequent clinical manifestations. Hematological evaluation revealed moderate to severe microcytic hypochromic anemia with elevated red blood cell distribution width (RDW). Comparative analysis demonstrated significantly lower hemoglobin, packed cell volume (PCV), mean corpuscular hemoglobin (MCH), and mean corpuscular hemoglobin concentration (MCHC) values in HbE-β-thalassemia patients, whereas RDW was significantly higher (p < 0.05). HPLC analysis showed significantly elevated HbF and HbA₂ levels in HbE-β-thalassemia. No significant correlation was observed between HbF level and hemoglobin concentration (r = -0.087, p = 0.474). Conclusion Double heterozygous hemoglobinopathies are an important cause of chronic hemolytic anemia in eastern India, particularly among tribal populations with prevalent consanguinity. HbE-β-thalassemia showed relatively more severe anemia and anisocytosis compared with Sβ-thalassemia. HPLC proved to be a reliable tool for the accurate characterization of hemoglobin variants and compound heterozygous states. Early diagnosis, targeted screening, and genetic counseling may help reduce disease burden in high-risk populations.