MULTIPLEX MUTAGENICALLY SEPARATED PCR - DIAGNOSIS OF BETA-THALASSEMIAAND HEMOGLOBIN-VARIANTS

A rapid and simple method, termed multiplex mutagenically separated PC R (MS-PCR), was developed to detect several molecular defects in the h emoglobin gene in one PCR. This technique, in which different-size all ele-specific primers were used, specifically amplified both normal and mutant alleles of the globin gene in the same reaction. Subsequent ge l electrophoresis showed at least one of the two allelic products at t he same locus or two of the several allelic products of different locc i and provided a within-assay quality control for the exclusion of fal se-negative results. In our study, the four most common beta-thalassem ia mutations, together with four other common hemoglobin variants in C hinese, were tested. Using multiplex MS-PCR 6 to 12 primers were added simultaneously into on reaction tube to identify one to four mutation s. Not only is this multiples MS-PCR method reliable and non-isoptic, the results can be obtained in less than one working day.