Determination of mutations in iron regulating genes of beta thalassemia major patients of Khyber Pakhtunkhwa, Pakistan

Abstract

Background Hepcidin and hemochromatosis (HFE) are iron regulatory proteins that are encoded by HAMP and HFE genes. Mutation in either HAMP gene or HFE gene causes Hepcidin protein deficiency that can lead to iron overload in beta thalassemia patients. The aim of this research work was to study the presence of G71D mutation of HAMP gene and H63D mutation of HFE gene in beta thalassemia major and minor group to check the association of these mutations with serum ferritin level of beta thalassemia patients. Methods The study was conducted on 42 beta thalassemia major and 20 beta thalassemia minor samples along with 20 control samples. The genotyping of both mutations has done by ARM‐PCR technique with specific set of primers. Results Significant effect of G71D and H63D mutations was observed on serum ferritin level of thalassemia major group. The risk allele of HAMP G71D and HFE H63D was found with high frequency (48% and 49%, respectively) in beta thalassemia major than in control group. High genotypic frequency of HAMP and HFE gene mutation gene mutation was observed in beta thalassemia major than beta thalassemia minor and control group (7% and 9%, respectively). Conclusion It can be concluded that both HAMP and HFE gene mutations show high frequency in beta thalassemia major patients and mean significant association between mutations and high serum ferritin level of beta thalassemia major patients but the nonsignificant results of Odd ratios showed that both mutations do not act as major risk factor in beta thalassemia major.

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“…DNA was extracted from the whole blood using salting out (non-enzymatic) method and SNP genotyping was confirmed via T-ARMS PCR, previously adopted in our lab [ 25 , 26 ]. Specific primers were designed using primer blast software for each SNP ( Table 1 ).…”

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“…Two sets of outer and inner primers (forward and reverse) were used to detect homozygous and heterozygous mutations. Extraction of genomic DNA was carried out using non-enzymatic/salting out methods already adopted in our lab [12,13].…”

Association of candidate genes (ALR2, RAGE, and VEGF) polymorphisms with diabetic retinopathy in type 2 diabetic patients of Khyber Pakhtunkhwa, Pakistan

“…All the samples were subjected to DNA extraction by salting out (non-enzymatic) method, and SNP genotyping was done through T-ARMS PCR, previously adopted in our lab [11,12]. Speci c primers were designed through primer blast software for each SNP (Table 1).…”