SIMULTANEOUS SCREENING FOR BETA-THALASSEMIA MUTATIONS BY CHEMICAL CLEAVAGE OF MISMATCH

We used the chemical cleavage of mismatch (CCM) method to screen the β-globin gene simultaneously for Mediterranean β-thalassemia mutations. The β-globin gene was amplified in two segments encompassing the whole gene and hybridized to a corresponding labeled PCR product from a normal subject. All the known mutations tested were identified and discriminated. Three β-thalassemic subjects with previously undiagnosed mutations were identified as carriers of two rare DNA changes. The inheritance of the mutations could be traced in family studies, showing the reliability of the method even for prenatal diagnosis. The β-globin gene polymorphisms were also detected and the framework was determined for most alleles. Our results suggest further applicability of the CCM method as a means to screen a gene simultaneously for multiple mutations. © 1991.