Molecular profiling of thalassemia in Southeastern Romania.

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 -α3·7 deletion and anti-α3·7 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.