Detection of mutation in Codon 41/42 of Beta Thalassemia Patients Using a Nonradioactive Technique

Abstract

Probes labelled with the fluorescein dUTP3' - tail were used to detect the deletion at codon 41/42 of beta globin gene in beta thalassaemia patients. Most of the mutations at codon 41/42 were previously described in Chinese or Asian Indians. The method includes Polymerase Chain Reaction (PCR) using two specific primers to amplify the 1.4kb region containing exon 1 and exon 2 of the human beta globin gene. This was followed by dot - blot hybridization and detected with a nonradioactive method based on the Enhanced Chemiluminescein (ECL) labelled oligonucleotides probes. Three concentrations of probes (5, 10, 15 mg/ml) were studied in this project. With the probes, we studied 44 beta thalasseamia chromosomes from 44 unrelated Malaysian patients. From our results, the probe for 4 bp deletion at codon 41/42 detected 16 patients, with 6 patients being homozygous and 10 were heterozygous. The most suitable concentration range of probe was 5-10 mg/ml.