title: Extensive sequencing of the cystic fibrosis transmembrane regulator gene: assay validation and unexpected benefits of developing a comprehensive test.
abstract: OBJECTIVE
  To develop a sequencing assay for the gene to identify mutations in patients with cystic fibrosis (CF).
  
  
  METHODS
  An automated assay format was developed to sequence all exons and splice junctional sequences, the promotor region, and parts of introns 11 and 19.
  
  
  RESULTS
  After validating the assay using 20 known samples, DNA of seven patients, four of whom were heterozygous for a known CF mutation, was sequenced. Known CF mutations were detected in seven of the eight chromosomes, and a novel missense mutation was detected in the eighth. In addition, this assay allowed 14 ambiguous results obtained using the Roche CF gold strips to be resolved. Three false-positive diagnoses were prevented; a different mutation at the same codon was identified in two patients and confirmation was provided in the remaining nine cases.
  
  
  CONCLUSIONS
  Sequencing of the gene provides important information for CF patients and is a valuable adjunct to a carrier screening program to resolve ambiguities in panel testing.
authors.affiliation: Molecular Genetics Laboratory, Quest Diagnostics Nichols Institute, San Juan Capistrano, California 92690, USA.
authors.forename: Charles M
authors.initials: CM
authors.lastname: Strom
authors.forename: Donghui
authors.initials: D
authors.lastname: Huang
authors.forename: Christina
authors.initials: C
authors.lastname: Chen
authors.forename: Arlene
authors.initials: A
authors.lastname: Buller
authors.forename: Mei
authors.initials: M
authors.lastname: Peng
authors.forename: Franklin
authors.initials: F
authors.lastname: Quan
authors.forename: Joy
authors.initials: J
authors.lastname: Redman
authors.forename: Weimin
authors.initials: W
authors.lastname: Sun
mesh_terms: D003062:Codon / Q000235:genetics; D003550:Cystic Fibrosis / Q000175:diagnosis / Q000453:epidemiology / Q000235:genetics*; D019005:Cystic Fibrosis Transmembrane Conductance Regulator / Q000235:genetics*; D004247:DNA / Q000235:genetics; D004252:DNA Mutational Analysis / Q000379:methods*; D005091:Exons; D005820:Genetic Testing; D006801:Humans; D007438:Introns; D008969:Molecular Sequence Data; D009154:Mutation
chemical_list: C505032:CFTR protein, human; D003062:Codon; D019005:Cystic Fibrosis Transmembrane Conductance Regulator; D004247:DNA