Rapid Detection of Six Common Mediterranean and Three Non-Mediterranean α-Thalassemia Point Mutations by Reverse Dot Blot Analysis

Abstract

We describe the implementation of reverse dot blot (RDB) hybridization as a rapid nonradioactive method for the identification of six frequent globin gene point mutations in the Mediterranean population: α Hphα: α2 IVS I donor site GGTGAGG → GG-; αNcolα: α2 initiation codon ATG → ACG; αTSaudlα: α2Poly A signal AATAA → AATAAG; αIcarlaα: α2 termination codon TAA → AAA (Ter → LYS); αCSα: α2 termination codon TAA → CAA (Ter → gly); ααNcol: α1 initiation codon ATG → GTG; and three α2 globin gene point mutations found in immigrants in Italy: αT-Quongszeα: α2 codon 12 CTG → CCG (Leu → Pro); αSeal Rockα: α2 termination codon TAA → GAA (TER → GLU); and α Koyadoraα: α2 termination codon TAA → TCA (TER → SER). The method uses the principle of allele-specific oligonucleotide (ASO) hybridization, but it is a nonradioactive method and permits rapid and simultaneous typing of point mutations and small deletions. © 2003 Wiley-Liss, Inc.