Abstract
Many polymerase chain reaction (PCR)-based techniques have been used for routine diagnosis of α- and β-thalassemias. However, most require a multi step of post-PCR processes that are time-consuming and labor-intensive procedures. This study reported the successful use of multiplex quantitative real-time PCR (qPCR), with high-resolution melting (HRM) analysis for diagnosis of two common deletional α0-thalassemia (α0-thal) and 15 common β-thalassemia (β-thal) mutations, in order to identify a couple at-risk of having a newborn with severe thalassemia in the northern region of Thailand. With this approach, 22 (7.2%) of 306 couples were diagnosed as being at-risk for having a child with severe thalassemia, including three homozygous α0-thal, five homozygous β-thal and 14 Hb E (HBB: c.79G>A)/β0-thal disease. Our findings indicated that multiplex qPCR with HRM is applicable for routine molecular diagnosis in order to identify a couple at-risk of having a newborn with severe thalassemia, especially in an endemic region.
Acknowledgements
The authors would like to thank medical technicians and nurses at the 15 district hospitals for their help and assistance.
Author contributions
C. Ruengdit and M. Punyamung performed all the experiments. C. Ruengdit, M. Punyamung, P. Khamphikham, P. Pongpunyayuen, N. Intasai and S. Pornprasert analyzed data and interpreted the results. C. Ruengdit, P. Khamphikham and S. Pornprasert wrote the manuscript. S. Pornprasert conceptualized the research idea. All authors have read and approved the final manuscript.
Disclosure statement
The authors report no conflicts of interest. The authors alone are responsible for the content and writing of this article.
Data availability
The data that support the findings of this study are available from the corresponding author upon reasonable request.