Detection of IVSII-1 mutation of Beta globin gene in patients with thalassemia minor using high-resolution melting analysis

Abstract

Background: 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 β-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 patients carrying IVSII-1 (G/A) mutations from patients who do not have this mutation. Methods: In this study, blood samples collected from 30 patients with 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. Findings: High-resolution melting analysis identified patients carrying IVSII-1 (G/A) mutation with a sensitivity and specificity of 100. Conclusion: 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. © 2016, Isfahan University of Medical Sciences(IUMS). All rights reserved.