title: Rapid genotyping of known mutations and polymorphisms in beta-globin gene based on the DHPLC profile patterns of homoduplexes and heteroduplexes.
abstract: BACKGROUND
  Beta-thalassemia represents a great heterogeneity as over 200 mutations have been identified for the beta-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.
  
  
  METHODS
  We have performed the validation of a DHPLC assay for direct genotyping of known causative mutations in beta-globin gene using the chromatographic pattern-based strategy under partially-denaturing conditions.
  
  
  RESULTS
  DHPLC 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 beta-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.
  
  
  CONCLUSIONS
  This simple method can meet the requirements of direct genotyping of known beta-thalassemia mutations and/or polymorphisms in the clinical setting for Chinese and in general as a model for other populations.
authors.affiliation: Department of Medical Genetics, School of Basic Medical Sciences, Southern Medical University, Guangzhou 510515, Guangdong, PR China.
authors.forename: Qiang
authors.initials: Q
authors.lastname: Li
authors.forename: Li-Yan
authors.initials: LY
authors.lastname: Li
authors.forename: Sheng-Wen
authors.initials: SW
authors.lastname: Huang
authors.forename: Liang
authors.initials: L
authors.lastname: Li
authors.forename: Xiao-Wei
authors.initials: XW
authors.lastname: Chen
authors.forename: Wan-Jun
authors.initials: WJ
authors.lastname: Zhou
authors.forename: Xiang-Min
authors.initials: XM
authors.lastname: Xu
mesh_terms: D002851:Chromatography, High Pressure Liquid / Q000379:methods; D004252:DNA Mutational Analysis / Q000379:methods; D005838:Genotype; D005914:Globins / Q000235:genetics*; D020180:Heteroduplex Analysis / Q000379:methods*; D006801:Humans; D009154:Mutation; D011110:Polymorphism, Genetic; D013997:Time Factors; D017086:beta-Thalassemia / Q000175:diagnosis / Q000235:genetics
chemical_list: D005914:Globins