Usage of Nondradioactive Probes Directly Labelled with Horseradish Peroxidase in the Diagnosis of §-Thalassemia

Authors: Hüseyin BAĞCI

Abstract: We describe a new, simple, rapid and sensitive nonradioactive technique for the analysis of genetic variations. Genomic DNA was amplified using polymerase chain reaction (PCR) and amplified DNA was hybridized with horseradish peroxidase (HRP-EC 1.11.1.7)-labelled allele specific oligonucleotide (ASO) probes. High specificity and sensitivity was achieved when labelling the ASO probes with colorimetric detection of HRP according to the 5'thiol oligolabelling system. This method was applied to the analysis of §-thalassemia common mutations in the Mediterranean region in PCR amplified genomic DNA and resulted in highly specific, sensitivesignals discriminating even in the cases of a one pair mismatch. This technique is particullarly convenient for §-thalassemia screening programme in Turkey because of allowing the use of nonradioctive chemicals for the simplification of hybridization and washing conditions.

Keywords: Polymerase Chain Reaction Horseradish peroxidase, §-thalassemia, Nonradioactive detection, 0ligonucleotide probes