Molecular profiling of thalassemia in Southeastern Romania

Abstract

Abstract Background Thalassemias represent a major group of hereditary hemoglobin disorders with significant global health impact, yet molecular data from Eastern European populations, particularly from Romania, remain limited. Purpose This study was aimed at comprehensively characterizing the molecular spectrum of α-, β-, and δβ-thalassemia in the southeastern part of Romania using an expanded panel of molecular diagnostic techniques. Methods A total of 324 patients with suspected thalassemia were evaluated between 2017 and 2025. Molecular testing included multiplex PCR with reverse dot-blot hybridization, Sanger sequencing, MLPA for α-globin deletions/duplications, and multiplex gap-PCR for δβ fusion gene detection. Results Pathogenic variants were identified in 137 individuals (42.28%). Of these, 21.89% had α-thalassemia, 66.42% had β-thalassemia, 10.21% had combined α and β defects, and 1.45% had Lepore Hb variants. The −α 3 · 7 deletion and anti-α 3 · 7 triplication were the predominant α-globin abnormalities, while IVS I-6 [T > C], IVS I-110 [G > A], codon 39 [C > T], and IVS II-745 [C > G] were the most frequent β-globin mutations. Two cases of the Hb Lepore Boston-Washington subtype were confirmed through gap-PCR and sequencing. Conclusions The mutation distribution observed in southeastern Romania mirrors Mediterranean thalassemia patterns rather than those of Central or Northern Europe.