Detection of IVSII-1 Mutation of Beta Globin Gene in Carriers of Thalassemia Minor Using High-Resolution Melting Analysis

Beta-thalassemia is one of the most common autosomal recessive disorders in the world population caused by more than 200 different mutations in the beta-globin chain. It is clinically classified as minor, intermediate and major. Beta-thalassemia is the most common monogenic disease in the Mediterranean countries, Middle East, Indian Subcontinent, and Southeast Asia and one of the widespread hereditary disorders in Iran. Among different
-thalassemia mutations identified among Iranian populations, IVSII-1 (G/A) is the most frequent mutation in different regions of Iran. This study aimed to determine the specificity and sensitivity of high-resolution melting (HRM) method in the diagnosis of individuals carrying IVSII-1 (G/A) mutations from patients who do not have this mutation. In this study, blood samples collected from 30 individuals carrying minor thalassemia were assessed. The genotype of each sample was previously determined via the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) or amplification-refractory mutation system (ARMS) or sequencing method in the genetic laboratory of Al-Zahra hospital, Isfahan, Iran. DNA extraction from peripheral blood was performed and high-resolution melting method was used for genotype samples. The results were analyzed according to the normalized and difference plots. High-resolution melting analysis identified individuals carrying IVSII-1 (G/A) mutation with a sensitivity and specificity of 100%. In summary, high-resolution melting method showed high sensitivity and specificity. Therefore, it is an appealing technique for identification of common mutations in genetic diseases.