Diagnosis of α 0 -thalassemia Chiang Rai (-- CR ) deletion by melt curve analysis in Northern Thailand.

A large novel 44.6 kb deletion named α 0 -thalassemia Chiang Rai (-- CR ) was first described in the individuals with uncommon Hb Bart's hydrops fetalis and HbH disease. This study aimed to develop a real-time gap PCR and melt curve analysis for the detection of -- CR and investigate its frequency in northern Thailand. Among 4,952 blood samples, the assay was performed in 525 samples with a mean corpuscular volume (MCV) < 80 fL, HbA 2 < 3.5%, HbA 2 +E < 25%, and negative for common deletional α 0 -thalassemia -- SEA and -- THAI . The developed method showed T m values of 85.8 ± 0.0 °C and 91.5 ± 0.1 °C, which were specific for -- CR and wild-type alleles, respectively. Nine (0.18% of 4,952 or 1.71% of 525) were positive for -- CR , in which two were HbH disease and the rest were heterozygous for -- CR . This study demonstrated the success of real-time gap PCR with melt curve analysis for -- CR diagnosis. Additionally, the prevalence of -- CR in the northern Thai population was comparable to -- THAI . Thus, this study implies the importance of -- CR in northern Thailand. Moreover, the developed real-time gap PCR with melt curve analysis is simple and highly accurate, and may be considered as an additional tool for routine α 0 -thalassemia -- CR diagnosis in this region.