Clinical and molecular diagnosis of betathalassemia in sample of iraqi patients

The purpose of this thesis is to investigate the molecular parameters in the beta thalassemia diagnosis and use the molecular techniques for fast and accurate diagnosis. Between January and April 2022, 150 patients were identified with beta thalassemia in Azadi Teaching Hospital and Al Jumhuri Hospital. Private laboratories in Kirkuk, Iraq, conducted experimental work. The volunteers in current study were divided to 50 healthy volunteers as control group. 100 beta thalassemia patients as a second group. ARMS-PCR device permits the detection of point mutations directly by the absence or presence of amplification using primers of specific allele. For the characterization of specific point mutation, a pair of allele-specific primers one of which has its 3′ terminal nucleotide complementary to the point mutation (Mt ARMS primer) and other to the normal DNA sequence (NARMS primer) was used. According to the distribution of common of β-thalassemia mutations in the surrounding countries, two mutations are selected for molecular characterization. The current results of RBC count was (P <0.05) in patients compared to control group. Hemoglobin concentration (P <0.05) in patients compared to control group PCV (P <0.05) patients compared to control group MCV and WBC demonstrated non-significant (P <0.05) changes in patients compared to control group. Hepcidin concentration (P <0.05) elevated in patients compared to control group. Ferritin concentration (P <0.05) elevated in patients compared to control group. Molecular analysis revealed that the 100 beta thalassemia cases studied for mutational pattern, IVS 1-5 (G→C) was present in 3(3%) among them. In all successful ARMS-PCR reactions, the internal control product of 861bp molecular weight was observed, which was considered as a mandatory sign of successful reaction upon gel electrophoresis. So out of the 100 cases studied for beta thalassemia mutational pattern, in 3 samples (3%), the mutational pattern IVS 1-5 (G→C) was detected. The rest 97 samples (97%) were negative for the IVS 1-5 (G→C) beta thalassemia mutation.