تشخيص جهش هاي حذفي Hb Lepore و Sicilian Hb در ژن بتاگلوبين با روش تكثير هم دما

Introduction: Thalassemia is one of the most common single-gene genetic disorders, caused by defects in the synthesis of hemoglobin molecule chains, and includes two types: alpha and beta. Beta thalassemia is a type of thalassemia caused by mutations in the beta-globin gene, which plays a role in the production of the beta chain of the hemoglobin molecule. This gene is located on chromosome 11 and is inherited in an autosomal recessive manner. Thalassemia can be caused by large deletion mutations located on the beta-globin gene cluster. Various deletion mutations have been identified in different populations. Two deletion mutations, Hemoglobin Lepore and Sicilian, are among the deletion mutations of beta thalassemia. Hemoglobin Lepore is a small group of structurally abnormal hemoglobins resulting from the fusion between the 5’ end of the delta-globin gene and the 3’ end of the beta-globin gene due to crossover or gene conversion during meiosis, leading to a 4.7 kb deletion between the delta and beta-globin genes. The Sicilian deletion mutation is a 13.4 kb deletion from the IVS II region of the delta-globin gene from the L1 element to the 3’ end of the beta-globin gene. Various molecular methods have been used to detect point mutations in beta thalassemia. Currently, many PCR-based methods can be used to identify deletion mutations in the globin gene, including Gap-PCR, Real-time PCR, and MLPA Methods: In this research project, primer sets were separately designed for deletion mutations in the Hemoglobin Lepore and Sicilian genes. The sensitivity and specificity of the designed specific primers were measured by performing isothermal amplification tests using a real-time thermal cycler, and the results were confirmed by gel electrophoresis. The feasibility of rapid testing using SYTO dye was eva‎luated. The effect of amorphous silica nanoparticles and zeolite nanoparticles on increasing the stability of the reaction mixture was investigated. Results and Discussion: The use of loop-mediated isothermal amplification (LAMP) for the detection of deletion mutations in the Hemoglobin Lepore and Sicilian genes was successful. The positive effect of silica nanoparticles in increasing the stability of the reaction mixture was observed.

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سيد حسين جلالي