DETECTION OF BETA-THALASSEMIA MUTATIONS USING DNA HETERODUPLEX GENERATOR MOLECULES

In this report we describe a rapid polymerase chain reaction (PCR) bas ed method for the detection of beta-thalassaemia (beta-thal) mutations . This method is based on the visualization of unique DNA heteroduplex banding patterns, following non-denaturing polyacrylamide gel electro phoresis, resulting from hybridization between mutant PCR products and synthetic DNA heteroduplex generator molecules. Using the Singaporean population, which consists of Chinese, Malay and Asian Indian ethnic groups, as a model, we have constructed and evaluated three DNA hetero duplex generator molecules for the detection of the common beta-thalas saemia mutations found in this population, The results show that these three molecules are capable of detecting approximately 95% of the mut ations found in the Singaporean population. We propose that this techn ology may be applied as an alternative screening strategy for beta-tha lassaemia mutations because it is technically simple, flexible, cost-e ffective, and requires only minimal laboratory resources.