Genotype–Phenotype Correlation of β-Thalassemia in the Indian College Student Population: Insights and Challenges in Screening and Molecular Diagnosis

ABSTRACT: Thalassemia, a widespread global health issue stemming from abnormal haemoglobin levels, affects approximately 4.5% individuals worldwide. Despite advances in treatment, this study investigates β-thalassemia among 3,000 college students from West Bengal, India, examining genotype-phenotype correlations and silent carrier prevalence. Methodologically, blood analyses and DNA extraction were used to screen participants. PCR amplification of four primers covering the HBB gene, followed by amplicon purification and nucleotide sequencing, was employed. Bioinformatics tools, evolutionary conservation analysis, and machine learning-assisted variant categorization were utilized to provide insights for clinicians. Results revealed 385 thalassemia carriers among 2,984 individuals, exhibiting various traits including β-thalassemia, haemoglobin E-heterozygotes, haemoglobin D Punjab heterozygotes, hereditary persistent foetal haemoglobin (HPFH), and borderline βthalassemia traits. Significant phenotypic variations were observed. Genotype analysis identified six mutations, each associated with distinct ethnic prevalences and clinical presentations. The discussion addressed diagnostic challenges, emphasizing the need for accurate diagnosis through haemoglobin analysis and DNA testing. Pathogenicity assessments provided insights into mutation impact. Overall, this study contributes to β-thalassemia management by offering guidance for research and clinical practice.