APPLICATION OF REAL-TIME PCR FOR RAPID DETECTION OF TWO ß-GLOBIN GENE MUTATIONS FROM THE PATIENTS WITH THALASSEMIA SYNDROMES

Abstract

Two mutations that have been found very frequently in Asian p-thalassemia patients were 4 base pair-TCTT deletion in codon 41 - 42 and A to T substitution in codon 17. In order to quickly detect these mutations in Vietnamese patients, we applied a Real-time PCR on the LightCycler 2.0 system with two sets of hybridization probe, which are specific for these mutations. Sixteen DNA samples extracted from blood of the children with j3-thalassemia syndromes were used as templates for mutation detection by using LightCycler Kit. The results obtained by Melting Curve Analysis showed that of these DNA samples, except one damaged during DNA extraction, four samples carried CD 17 heterozygous mutation, five samples carried CD41142 homozygous mutation, one sample carried both of the mutations and the last five samples did not carry any mutations. From these results, we concluded that frequency of two mutations at 17 and 41/42 codons is also high in the Vietnamese p-thalassemia patients