Molecular characterization of similar Hb Lepore Boston-Washington in four Chinese families using third generation sequencing

• Published in: Scientific reports Vol. 14; no. 1; pp. 9966 - 8
• Main Authors: Zhuang, Jianlong, Zhang, Na, Zheng, Yu, Jiang, Yuying, Chen, Yu’e, Mao, Aiping, Chen, Chunnuan
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 30-04-2024; Nature Publishing Group; Nature Portfolio
• Subjects: 631/208; 692/700; Adult; Asian People - genetics; beta-Globins - genetics; beta-Thalassemia - blood; beta-Thalassemia - diagnosis; beta-Thalassemia - genetics; Blood diseases; Capillary electrophoresis; China; Diagnosis; East Asian People; Female; Fusion protein; Gap-PCR; Gene fusion; Genes; Genetic screening; Genetic testing; Hb Lepore; Hemoglobin; Hemoglobins, Abnormal - genetics; Heterozygotes; High-Throughput Nucleotide Sequencing - methods; Humanities and Social Sciences; Humans; Hybridization; Male; multidisciplinary; Pedigree; Polymerase chain reaction; Sanger sequencing; Science; Science (multidisciplinary); Thalassemia; Third-generation sequencing; China; Gap-PCR; Thalassemia; Third-generation sequencing; Sanger sequencing; Hb Lepore
• ISSN: 2045-2322; 2045-2322
• DOI: 10.1038/s41598-024-60604-7

Hemoglobin (Hb) Lepore is a rare deletional δβ-thalassemia caused by the fusion between delta-beta genes, and cannot be identified by traditional thaltassemia gene testing technology. The aim of this study was to conduct molecular diagnosis and clinical analysis of Hb Lepore in four unrelated Chinese families using third generation sequencing. Decreased levels of mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH) and an abnormal Hb band were observed in the probands of the four families. However, no common α and β-thalassemia variants were detected in the enrolled families using polymerase chain reaction-reverse dot blot hybridization based traditional thalassemia gene testing. Further third-generation sequencing revealed similar Hb Lepore-Boston-Washington variants in all the patients, which were resulted from partial coverage of the
HBB
and
HBD
globin genes, leading to the formation of a delta-beta fusion gene. Specific gap-PCR and Sanger sequencing confirmed that all the patients carried a similar Hb Lepore-Boston-Washington heterozygote. In addition, decreased levels of MCH and Hb A2 were observed in the proband’s wife of family 2, an extremely rare variant of Hb Nanchang (GGT > AGT) (HBA2:c.46G > A) was identified by third-generation sequencing and further confirmed by Sanger sequencing. This present study was the first to report the similar Hb Lepore-Boston-Washington in Chinese population. By combining the utilization of Hb capillary electrophoresis and third-generation sequencing, the screening and diagnosis of Hb Lepore can be effectively enhanced.