[Analysis of beta thalassemia mutations using the single strand conformation polymorphism [SSCP] technique]

beta-thalassemia [beta-thal] is one of the most prevalent hereditary diseases in Iran. There are more than two million carriers of beta-thal in Iran. Detection of the beta globin gene mutations is necessary for a definitive diagnostic and management plan such as prenatal diagnosis of beta-thalassemia. In our country, the PCR-Amplification Refractory Mutation System [PCR-ARMS] has been frequently used for detection of beta globin gene mutations. Here, we used the PCR-single strand conformation polymorphism [PCR-SSCP] assay for detection of mutations of beta globin gene. In the patients with confirmed mutations, we amplified 281base pairs containing exon of one of a beta globin gene by PCR. Based on SSCP technique 2.5 micro l of the reaction products appeared in polyacryamide gel electrophoresis and the bands were visualized by silver staining. Seven mutations and one polymorphism were evaluated by PCR SSCP assay. The results of this study demonstrated that the patterns of mobility of single strands were different from each other and those of control sample. Our study showed the PCR-SSCP technique can meet the need for direct genomic sequencing of DNA and could be applied in the developing countries where financial resources are limited but genetic hemoglobin disorders are highly prevalent

[Detection of beta-thalassemia mutations in Kurd patients of Kurdistan and West Azerbaijan provinces]

Beta-thalassemia is the most common autosomal recessive disorder. More than 200 different mutations in the beta-globin gene have been detected which can lead to decreased or absent beta-globin chain synthesis. Since the Iranian Population is a mixture of different ethnic groups, it is necessary to determine the frequency and distribution of these mutations in the different ethnic groups of our county. Therefore, in this study we determined the Spectrum and the frequency of beta-thalassemia mutations in the patients with beta-thalassemia major in the Kurd population of Kurdistan and West Azerbaijan provinces of Iran. To detect mutations, extracted DNA of 110 chromosomes from 55 unrelated patients, were studied by PCR-ARMS [Polymerase Chain Reaction-Amplification Refractory Mutation System] SSCP [Single Strand Conformation Polymorphism] and direct sequencing methods. The results of this study showed that IVS-II-1 [G-A] was the most common mutation with a frequency of 31%; FSC 8/9[+G] with a frequency of 19% was the second most prevalent mutation among all chromosomes. Other mutations were IVS-I-1[G-A] FSC8 [-AA] IVS-I-110[G-A] FSC36/37[-T] IVS-I-5[G-C], IVS-I-128[T-G] FSC44 [-C], FSC 5[-CT] and +22UTR [G-A] These mutations comprised 79% of beta-thalassemia mutations in this region and 21% of the mutations still remains to be explored. The results of this study showed that, there are similarities and differences between this region and other parts of Iran and also neighboring countries. Therefore, determination of beta-thalassemia mutations in this region seems to be necessary and beneficial for designing prenatal diagnosis programs