Hemoglobin profile and molecular characteristics of the complex interaction of hemoglobin Doi-Saket [α9(A7) asn > lys, HBA2:c.30C > a], a novel α2α1 hybrid globin variant, with hemoglobin E [β26(B8) Glu > lys, HBB:c.79G > A] and deletional α+-thalassemia in a Thai family

Background An increasing number of alpha-hemoglobin (Hb) variants is causing various clinical symptoms; therefore, accurate identification of these Hb variants is important.Objective This study aimed to describe the molecular and hematological characteristics of novel Hb Doi-Saket that gives rise to a typical alpha+-thalassemia phenotype in carriers with and without other hemoglobinopathies.Materials and Methods Biological samples from a proband and his family members were analyzed. Hematological profiles were analyzed using a standard automated cell counter. Hb was analyzed by capillary electrophoresis and high-performance liquid chromatography. Mutations and globin haplotype were identified by DNA analysis. Novel diagnostic tools based on allele-specific polymerase chain reaction (PCR) and PCR-restriction fragment length polymorphism were developed.Results Hb analysis showed a major abnormal Hb fraction, moving slower than HbA, and a minor Hb fraction alongside HbA2 in the proband, his father, and son. DNA analysis of the alpha-globin gene identified the -alpha(3.7) deletion and in cis the C > A mutation on codon 9 of the alpha 2 alpha 1 gene, corresponding to Hb Doi-Saket [alpha 9(A7) Asn > Lys]. This mutation could be identified using newly developed allele-specific PCR-based assays. The Hb Doi-Saket al.lele was significantly associated with haplotype [- + M + + 0 -]. Interaction of alpha Doi-Saket with beta E globin chains led to a new Hb variant (HbE Doi-Saket). Phenotypic expression was clinically silent in heterozygotes and might present slight microcytosis.Conclusions Hb Doi-Saket emphasizes a great diversity present in alpha-globin gene. The mutation in this family from Thailand was linked to -alpha(3.7) and caused mild microcytosis in the carriers. The combination of this variant with deletions in alpha genes might cause a severe clinical phenotype. Different methods of separation can provide useful information in diagnosis, and a complete molecular approach is needed for confirmation before considering patient management.