title: Detection of large rearrangements in the cystic fibrosis transmembrane conductance regulator gene by multiplex ligation-dependent probe amplification assay when sequencing fails to detect two disease-causing mutations.
abstract: OBJECTIVE
  Most of the over 1600 mutations and sequence variants identified to date in the cystic fibrosis transmembrane conductance regulator (CFTR) gene are point mutations or small deletions/insertions detectable by conventional sequencing. However, large rearrangements (deletions, duplications, or insertion/deletion mutations) have recently been reported to constitute 1-2% of CFTR mutations. The CFTR sequencing protocol at ARUP Laboratories interrogates the coding regions of all 27 exons and all intron/exon boundaries of the gene. This study was undertaken to determine whether testing for large gene rearrangements could improve the mutation detection rate.
  
  
  RESULTS
  Nine cases with abnormal quantitative pilocarpine iontophoresis sweat chloride (SC) values (>60 mEq/L) and 20 cases with borderline SC levels (40-60 mEq/L) with only one or no mutations detected by the ARUP 32 mutation panel, including the 23 mutations recommended by American College of Medical Genetics (ACMG) for carrier screening, followed by sequencing, were tested using a multiplex ligation-dependent probe amplification (MLPA) assay. MLPA analysis identified one deletion among nine patients with SC >60 who had previously been tested with sequencing. None of the cases with borderline SC levels showed rearrangements.
  
  
  CONCLUSIONS
  The MLPA assay for detection of large rearrangements may be valuable in individuals with positive SC levels where one or no mutations have been identified by sequencing.
authors.affiliation: Department of Pathology, University of Utah, Salt Lake City, Utah, USA. asvensson@hsc.wvu.edu
authors.forename: Annika M
authors.initials: AM
authors.lastname: Svensson
authors.forename: Lan-Szu
authors.initials: LS
authors.lastname: Chou
authors.forename: Christine E
authors.initials: CE
authors.lastname: Miller
authors.forename: Jorge A
authors.initials: JA
authors.lastname: Robles
authors.forename: Jeffrey J
authors.initials: JJ
authors.lastname: Swensen
authors.forename: Karl V
authors.initials: KV
authors.lastname: Voelkerding
authors.forename: Rong
authors.initials: R
authors.lastname: Mao
authors.forename: Elaine
authors.initials: E
authors.lastname: Lyon
mesh_terms: D002712:Chlorides / Q000032:analysis; D003550:Cystic Fibrosis / Q000175:diagnosis* / Q000235:genetics*; D019005:Cystic Fibrosis Transmembrane Conductance Regulator / Q000235:genetics*; D004252:DNA Mutational Analysis / Q000379:methods; D005091:Exons; D015321:Gene Rearrangement; D006801:Humans; D007438:Introns; D009154:Mutation; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D013542:Sweat / Q000737:chemistry
chemical_list: C505032:CFTR protein, human; D002712:Chlorides; D019005:Cystic Fibrosis Transmembrane Conductance Regulator