Detection of Rare Thalassemia Variants Using Accurate Circular Consensus Long‐Read Sequencing

Objective The aim of this study is to evaluate the efficacy of accurate circular consensus long‐read sequencing in the detection of rare thalassemia. Methods Conventional molecular analysis on globin genes has limitations because of the broad spectrum of genetic variants, complex genetics, and genotype–phenotype correlation. Accurate circular consensus long‐read sequencing is a novel tool that detects complex variants in the thalassemia gene based on third‐generation sequencing. In this study, we screen out suspected rare thalassemia carriers by hemoglobin analysis and conventional molecular analysis, and evaluate the efficacy of accurate circular consensus long‐read sequencing in the detection of rare thalassemia. Results Based on the traditional screening of thalassemia gene, an additional 16 (17.67%) cases of clinically significant variants of rare thalassemia were identified by accurate circular consensus long‐read sequencing in this study, including 12‐point variants and 4 deletion variants: HBB: (SEA)‐HPFH, HBB: c.268_281delAGTGAGCTGCACTG, HBB: (Chinese) G γ + (A γδβ )0, and HBA2 :c.91‐93delGAG. Conclusion Accurate circular consensus long‐read sequencing has a promising prospect in detecting rare thalassemia gene variants and may improve the detection rate of carriers.