Rapid detection of deletions causing δβ thalassemia and hereditary persistence of fetal hemoglobin by enzymatic amplification

Abstract

A considerable number of deletions of variable size and position that involve the β-globin gene complex on chromosome 11 are associated with the clinical entities of hereditary persistence of fetal hemoglobin (HPFH) and δβ thalassemia. Specific deletions appear to be associated with consistent phenotypes and some are known to be recurrent. To facilitate the molecular diagnosis of uncharacterized patients with HPFH and δβ thalassemia, oligonucleotide primers have been designed to enzymatically amplify deletion- specific products for nine known deletions, which include those responsible for HPFH-1, HPFH-2, HPFH-3, Spanish (δβ)(O) thalassemia, hemoglobin (Hb) Lepore, Sicilian (δβ)(O) thalassemia, Chinese (G)γ((A)γδβ)(O) thalassemia, Asian-Indian inversion-deletion (G)γ((A)γδβ)(O) thalassemia, and Turkish inversion-deletion (δβ)(O) thalassemia. Using this approach, we have successfully characterized the molecular basis for δβ thalassemia in 23 individuals from 16 families of diverse ethnic origins. Thirteen individuals from this group were shown to be heterozygous for the 13.4-kb Sicilian deletion, two were heterozygous for the 100-kb Chinese (G)γ((A)γδβ)(O) deletion, four were heterozygous for the Turkish form of inversion-deletion δβ thalassemia, and three were heterozygous for the Asian-Indian form of inversion-deletion (G)γ((A)γδβ)(O) thalassemia. One Vietnamese subject was heterozygous for a 12.6-kb deletion, which we have fully characterized at the molecular level. Sequence analysis of the breakpoint regions of the Chinese deletion and the Turkish rearrangement indicates that, in each case, the mutation is likely to have arisen from a single origin. This hypothesis is supported by the evident geographical clustering of the various deletions described here.