Jg. Chang et al., RAPID DIAGNOSIS OF BETA-THALASSEMIA BY MUTAGENICALLY SEPARATED POLYMERASE CHAIN-REACTION (MS-PCR) AND ITS APPLICATION TO PRENATAL-DIAGNOSIS, British Journal of Haematology, 91(3), 1995, pp. 602-607
We have developed a rapid and simple PCR-based method which is modifie
d fi om the mutagenically separated polymerase chain reaction (MS-PCR)
to detect the molecular defects of beta-thalassaemia. We can use this
technique to amplify normal and mutant alleles of the beta-globin gen
e in the same reaction tube, using different-sized allele-specific pri
mers, This mutagenesis separates the amplification reactions of allele
s performed in the same tube. Subsequent gel electrophoresis shows at
least one of the two allelic products at the same locus or at least tw
o of the several allelic products at different loci. Therefore, in add
ition to simple handling, MS-PCR provides a within-assay quality contr
ol for the exclusion of false negative results. The five most common m
utations of beta-thalassaemia and haemoglobin E which occur in the Tai
wanese population were tested, and 14 prenatal samples were checked wi
th accurate results. This method is simple, rapid and accurate, and ca
n be used routinely in prenatal diagnosis. The principle used here can
also be applied to other genetic diseases.