title: Rapid detection of beta-globin gene mutations and polymorphisms by temporal temperature gradient gel electrophoresis.
abstract: BACKGROUND
  Inherited hemoglobin disorders represent the most common Mendelian disease worldwide. Prevention programs based on molecular diagnosis of heterozygous carriers and/or patients require the use of reliable mutation scanning methods in at-risk populations.
  
  
  METHODS
  We developed a rapid and highly specific mutation-screening test based on temporal temperature gradient gel electrophoresis (TTGE). We analyzed 889 beta-thalassemia genes from homozygous beta-thalassemia patients and unrelated individuals with heterozygous beta-thalassemia. Previously reported common mutations were screened by reverse dot blots using allele-specific probes. The rare mutations were analyzed by TTGE.
  
  
  RESULTS
  We found common mutations in 753 beta-thalassemia genes. TTGE analysis in the rest of the genes showed the presence of mutations in different regions of the beta-globin gene in 134 of them, and these mutations were characterized by DNA sequencing. In the two genes in which mutations were not identified, large deletions spanning beta-globin gene were suspected.
  
  
  CONCLUSIONS
  Compared with other approaches for comprehensive mutation screening, the reported method is rapid, highly sensitive, cost-effective, and suitable for high-throughput screening of a large number of samples.
authors.affiliation: Department of Haematology, Christian Medical College, Vellore 632004, India. rvshaji@cmcvellore.ac.in
authors.forename: Ramachandran V
authors.initials: RV
authors.lastname: Shaji
authors.forename: Eunice Sindhuvi
authors.initials: ES
authors.lastname: Edison
authors.forename: Balasubramanian
authors.initials: B
authors.lastname: Poonkuzhali
authors.forename: Alok
authors.initials: A
authors.lastname: Srivastava
authors.forename: Mammen
authors.initials: M
authors.lastname: Chandy
mesh_terms: D004591:Electrophoresis, Polyacrylamide Gel; D005914:Globins / Q000235:genetics*; D006801:Humans; D009154:Mutation; D009691:Nucleic Acid Denaturation; D016133:Polymerase Chain Reaction; D011110:Polymorphism, Genetic; D013696:Temperature
chemical_list: D005914:Globins