Detection of rare thalassemia mutations using long-read single-molecule real-time sequencing

•The conventional methods and Next-generation sequencing to diagnose thalassemia have limitations.•Long-read SMRT sequencing has been demonstrated to be more effective and accurate than conventional methods, especially for rare and complicated thalassemia variants.•A 762 bp deletion and a 342 bp insertion in α-globin gene cluster were identified by SMRT sequencing and reported for the first time.•Subjects with other rare mutations including α Fusion mutation, α-triplicates, α-quadruplicates and conversion of HBA2 to HBA1 was precisely identified by SMRT sequencing.