Diagnostic Utility of a Multiplex PCR Assay in Detecting Common Mutations of the α-Globin Gene in α-Thalassemia

Alpha-thalassemia is a hereditary hemoglobin disorder characterized by reduced or absent α-globin gene, and its severity is associated with the number of affected alleles. Several methods are available for detecting α-thalassemia, such as multiplex ligation-dependent probe amplification (MLPA) and PCR-based hybridization strip assay. Multiplex PCR offers a faster, more convenient, and cost-effective alternative. In this study, we aimed to optimize a current PCR-based method for α-thalassemia screening and evaluate its utility using clinical samples. We also investigated the prevalence and spectrum of common mutations responsible for α-thalassemia in Thailand and Korea. A total of 1261 samples from Thailand, 560 samples from different ethnic groups residing in Korea, and 300 samples from native Koreans were collected and tested. The concordance rate between the data collected in Thailand and in this study was 99.92%. Further, approximately 5.9% of the non-Korean individuals living in Korea were identified as healthy carriers, whereas no mutations were observed in Koreans. Comparing the data with MLPA or Sanger sequencing data showed 100% agreement rate in both cases. We successfully developed a PCR method for the diagnosis of α-thalassemia that is fast, less labor-intensive, and cost-effective. Given the performance results of this method, it has great potential for application in α-thalassemia diagnosis.