title: Application of pyrosequencing to the identification of sequence variations in the cystic fibrosis transmembrane conductance regulator gene.
abstract: BACKGROUND
  A high number of mutations associated with cystic fibrosis have been identified in the cystic fibrosis transmembrane conductance regulator (CFTR) gene, most of which are rare and, therefore, hamper extensive molecular diagnosis. In couples undergoing prenatal diagnosis where no mutation is found in one or both partners, additional analysis of intragenic polymorphisms may allow for the identification of fetal alleles associated with cystic fibrosis.
  
  
  METHODS
  We developed novel, rapid and accurate assays for CFTR genotype determination using pyrosequencing technology; a simple, automated and reliable technique with low cost.
  
  
  RESULTS
  Assays were optimized for the identification of the seven most frequent CFTR mutations (p.DeltaF508, p.N1303K, p.G542X, c.2183AA>G, c.1717-1G>A, p.W1282X, p.R1162X) in the Italian population and two common intragenic polymorphisms (rs213950 and rs1800136). Blind validation on 15 known control samples, typed for each sequence variation, allowed correct identification of all 135 genotypes.
  
  
  CONCLUSIONS
  We demonstrated that this procedure is highly specific for the identification of individual CFTR sequence variations associated with cystic fibrosis, allowing both population screening and prenatal diagnosis.
authors.affiliation: Dipartimento di Diagnostica, A.O.O.I.R.M.-S. Anna, Turin, Italy.
authors.forename: Caterina
authors.initials: C
authors.lastname: Mari
authors.forename: Francesca
authors.initials: F
authors.lastname: Bruno
authors.forename: Silvia
authors.initials: S
authors.lastname: Galbiati
authors.forename: Anna
authors.initials: A
authors.lastname: Torri
authors.forename: Federica
authors.initials: F
authors.lastname: Lombardo
authors.forename: Manuela
authors.initials: M
authors.lastname: Seia
authors.forename: Maurizio
authors.initials: M
authors.lastname: Ferrari
authors.forename: Gabriella
authors.initials: G
authors.lastname: Restagno
authors.forename: Laura
authors.initials: L
authors.lastname: Cremonesi
mesh_terms: D003550:Cystic Fibrosis / Q000175:diagnosis / Q000235:genetics; D019005:Cystic Fibrosis Transmembrane Conductance Regulator / Q000235:genetics*; D005260:Female; D005838:Genotype; D006801:Humans; D009154:Mutation; D011110:Polymorphism, Genetic; D011247:Pregnancy; D011296:Prenatal Diagnosis; D017422:Sequence Analysis, DNA / Q000379:methods*
chemical_list: D019005:Cystic Fibrosis Transmembrane Conductance Regulator