Rapid genotyping of known mutations and polymorphisms in β-globin gene based on the DHPLC profile patterns of homoduplexes and heteroduplexes

Backgroundβ-thalassemia represents a great heterogeneity as over 200 mutations have been identified for the β-globin gene responsible for this disease. A rapid genotyping test with high accuracy, selectivity, and reproducibility suitable for the determination of known mutations is needed for prenatal screening and post-natal diagnosis of this disease in clinical setting.Design and methodsWe have performed the validation of a DHPLC assay for direct genotyping of known causative mutations in β-globin gene using the chromatographic pattern-based strategy under partially-denaturing conditions.ResultsDHPLC assay was established based on the analysis of 795 DNA samples from a group of various genotypes for the 20 mutations and 8 polymorphisms in β-globin gene then validated on 319 tests in a blind study. The results obtained with this assay were in concordance with the results obtained by DNA sequence analysis.ConclusionThis simple method can meet the requirements of direct genotyping of known β-thalassemia mutations and/or polymorphisms in the clinical setting for Chinese and in general as a model for other populations.