Rapid detection of a-thalassaemia variants using droplet digital PCR

Introduction: Alpha thalassaemia is a highly prevalent disease globally and is a well-known public health problem in Malaysia. The deletional forms of the mutation are the most common forms found in alpha thalassacmia. The three most common deletional alpha thalassacmia found in this region include --(SEA) deletion, -alpha(3 7) rightward and -74 2 leftward deletions. The prevalence rate of triplication alpha cases such as alpha alpha alpha(anti3.7) and alpha alpha alpha(anti4.2) is not known in Malaysia although it plays a role in exacerbating the clinical phenotypes in beta thalassaemia carriers. Recently, there have been more reported cases of rare alpha thalassacmia mutations due to the advancement of molecular techniques involved in thalassacmia detections. Therefore, it is essential to develop a new method which allows the detection of different alpha thalassaemia minas including the rare ones simultaneously arid accurately. Methods: The purpose of this study was to design an assay for the detection of triplications, common and rare deletional alpha thalassacmia using droplet digital PCR (ddPCR). Results: This is a quantitative detection method to measure the changes of copy number which can detect deletions, duplications arid triplications of the alpha globin gene simultaneously. Conclusion: In conclusion, ddPCR is an alternative method for rapid detection of alpha thalassaemia variants in Malaysia.