title: Next generation diagnostics of cystic fibrosis and CFTR-related disorders by targeted multiplex high-coverage resequencing of CFTR.
abstract: BACKGROUND
  Here we have developed a novel and much more efficient strategy for the complete molecular characterisation of the cystic fibrosis (CF) transmembrane regulator (CFTR) gene, based on multiplexed targeted resequencing. We have tested this approach in a cohort of 92 samples with previously characterised CFTR mutations and polymorphisms.
  
  
  METHODS
  After enrichment of the pooled barcoded DNA libraries with a custom NimbleGen SeqCap EZ Choice array (Roche) and sequencing with a HiSeq2000 (Illumina) sequencer, we applied several bioinformatics tools to call mutations and polymorphisms in CFTR.
  
  
  RESULTS
  The combination of several bioinformatics tools allowed us to detect all known pathogenic variants (point mutations, short insertions/deletions, and large genomic rearrangements) and polymorphisms (including the poly-T and poly-thymidine-guanine polymorphic tracts) in the 92 samples. In addition, we report the precise characterisation of the breakpoints of seven genomic rearrangements in CFTR, including those of a novel deletion of exon 22 and a complex 85 kb inversion which includes two large deletions affecting exons 4-8 and 12-21, respectively.
  
  
  CONCLUSIONS
  This work is a proof-of-principle that targeted resequencing is an accurate and cost-effective approach for the genetic testing of CF and CFTR-related disorders (ie, male infertility) amenable to the routine clinical practice, and ready to substitute classical molecular methods in medical genetics.
authors.affiliation: Genetic Causes of Disease Group, Bioinformatics and Genomics Programme, Centre for Genomic Regulation (CRG), Doctor Aiguader 88, Barcelona, Catalonia 08003, Spain.
authors.forename: D
authors.initials: D
authors.lastname: Trujillano
authors.forename: M D
authors.initials: MD
authors.lastname: Ramos
authors.forename: J
authors.initials: J
authors.lastname: González
authors.forename: C
authors.initials: C
authors.lastname: Tornador
authors.forename: F
authors.initials: F
authors.lastname: Sotillo
authors.forename: G
authors.initials: G
authors.lastname: Escaramis
authors.forename: S
authors.initials: S
authors.lastname: Ossowski
authors.forename: L
authors.initials: L
authors.lastname: Armengol
authors.forename: T
authors.initials: T
authors.lastname: Casals
authors.forename: X
authors.initials: X
authors.lastname: Estivill
mesh_terms: D001483:Base Sequence; D001987:Bronchiectasis / Q000235:genetics; D015331:Cohort Studies; D003550:Cystic Fibrosis / Q000175:diagnosis* / Q000235:genetics*; D019005:Cystic Fibrosis Transmembrane Conductance Regulator / Q000235:genetics*; D005091:Exons; D005820:Genetic Testing; D006801:Humans; D007248:Infertility, Male / Q000235:genetics; D008297:Male; D008969:Molecular Sequence Data; D011110:Polymorphism, Genetic; D014649:Vas Deferens / Q000002:abnormalities
chemical_list: D019005:Cystic Fibrosis Transmembrane Conductance Regulator
keywords: Cystic fibrosis; Diagnostics; Genetic screening/counselling; Getting Research into Practice; Molecular genetics