Development of Molecular Techniques for Identification of β-Thalassemia Mutations

• Main Authors: Hung-Chang Shih, 施浤彰
• Other Authors: 張天傑
• Format: Others
• Language: zh-TW
• Published: 2009
• Online Access: http://ndltd.ncl.edu.tw/handle/82159874511741557697

博士 === 國立中興大學 === 獸醫學系暨研究所 === 97 === Thalassemia is clinically heterogeneous because various genetic lesions variably impair globin-chain synthesis. β-thalassemia is one of the most common genetic diseases in the world. It is an autosomal recessive inherited disease resulting from point mutations, small insertions, or deletions in the beta-globin (HBB) gene and the most common mutations of β-globin are point mutations. Over 700 mutations have been described in the beta-globin of which more than 200 are associated with β-thalassemia phenotype. In the Chinese population, more than 20 types of mutations have been found, therefore, the development of rapid, effective and economical techniques for screening and analysis of HBB mutations is essential. High resolution melting (HRM) analysis is a new gene scan tool that quickly performs the PCR and identifies sequences alterations without requiring post-PCR treatment. Herein we evaluated HRM analysis in the identification of HBB mutations. We have successfully established a diagnostic strategy for identifying HBB gene mutations including c.-78A>G, c.-79A>G, c.2T>G, c.79_80insT, c.84_85insC, c.84_85insC, c.123_124insT, c.125_128delTCTT, c.130 G>T, c.170G>A, c.216_217ins A and c.316-197 C>T from wild-type DNA using HRM analysis. The results of HRM analysis were confirmed by direct DNA sequencing. In summary, we report that HRM analysis is an appealing technique for the identification of HBB mutations. We also believe that HRM can be used as a method for prenatal diagnosis of β-thalassemia.