Comprehensive and efficient HBB mutation analysis for detection of β-hemoglobinopathies in a pan-ethnic population

Current methods that assay hemoglobin β-globin chain variants can have limited clinical sensitivity when applied techniques identify only a predefined panel of mutations. Even sequence-based assays may be limited depending on which gene regions are investigated. We sought to develop a clinically practical yet inclusive molecular assay to identify β-globin mutations in multicultural populations. We highlight the β-globin mutation detection assay ( β-GMDA), an extensive gene sequencing assay. The polymerase chain reaction (PCR) primers are located to encompass virtually all hemoglobin β locus (HBB) mutations. In addition, this assay is able to detect, by gap PCR, a common large deletion ( Δ619 base pair), which would be missed by sequencing alone. We describe our 5-year experience with the β-GMDA and indicate its capability for detecting homozygous, heterozygous, and compound heterozygous sequence changes, including previously unknown HBB variants. The β-GMDA offers superior sensitivity and ease of use with comprehensive detection of HBB mutations that result in β-globin chain variants.