Tetra - Primer ARMS PCR Optimization For Genotyping CD41/42 (-TTCT) Mutation On B-Globin Gene Associated With B-Thalassemia Disorder

Abstract

Thalassemia is an inherited blood disorder with two main types, α-thalassemia and β thalassemia. It is prevalent in various regions around the world including Mediterranean, Middle East, Asia-Pacific regions, and Vietnam. In which, β-thalassemia is caused by over 380 mutations on β-globin gene. Therefore, developing a method to screen for high-risk couples of β-thalassemia is extremely important for genetic counseling and controlling the new cases of β-thalassemia. In this study, the optimization of tetra-primer ARMS PCR to genotype one of the most common mutations of β-thalassemia in Vietname, CD41/42 mutation, was performed with saliva samples. Three criteria including annealing temperature, primers’ concentration and sample concentration were optimized to find the optimal condition for tetra-primer ARMS PCR assay. It has detected CD41/42 mutation and distinguished homozygous and heterozygous forms of the mutation, making it a potential option as a diagnostic test for β-thalassemia because it is not only non-invasive but also time-and-cost efficient compared with other conventional methods.