title: Evaluation of dHPLC in mutation screening of the APC gene in a Greek FAP cohort.
abstract: BACKGROUND
  Germline mutations in the APC gene predispose to colorectal adenomas leading to cancer in over 80% of patients. A multitude of mutations, dispersed throughout the gene, have been described. We wanted to evaluate the usefulness of denaturing high performance liquid chromatography (dHPLC) for mutation screening.
  
  
  METHODS
  Ten amplicons containing 14 mutations in the APC, previously identified by sequencing in 22 FAP patients, were analysed by dHPLC. dHPLC was also used to screen members of a family for a mutation identified in the proband.
  
  
  RESULTS
  We analysed 10 amplicons under a total of 59 temperatures. Successful results were obtained from 51 out of 59 tested temperatures (86.4%). In all cases a different heteroduplex-homoduplex pattern was obtained from mutant DNA in at least two of the temperatures. All 14 mutations identified by sequencing were also detected using dHPLC. Sequence analysis of a large family confirmed the dHPLC results.
  
  
  CONCLUSIONS
  Using dHPLC we detected all mutations previously identified by sequencing.
authors.affiliation: Molecular Biology Research Center HYGEIA, Antonis Papayiannis, Kifissias Ave. & Erythrou Stavrou 4 Str., 15123 Maroussi, Athens, Greece.
authors.forename: Markos
authors.initials: M
authors.lastname: Mihalatos
authors.forename: Angela
authors.initials: A
authors.lastname: Apessos
authors.forename: John K
authors.initials: JK
authors.lastname: Triantafillidis
authors.forename: Paris A
authors.initials: PA
authors.lastname: Kosmidis
authors.forename: George
authors.initials: G
authors.lastname: Fountzilas
authors.forename: Niki J
authors.initials: NJ
authors.lastname: Agnantis
authors.forename: Drakoulis
authors.initials: D
authors.lastname: Yannoukakos
authors.forename: George
authors.initials: G
authors.lastname: Nasioulas
mesh_terms: D011125:Adenomatous Polyposis Coli / Q000235:genetics*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D015331:Cohort Studies; D004273:DNA, Neoplasm / Q000032:analysis / Q000235:genetics; D017491:Genes, APC; D018095:Germ-Line Mutation; D006801:Humans; D010375:Pedigree; D016133:Polymerase Chain Reaction; D015203:Reproducibility of Results
chemical_list: D004273:DNA, Neoplasm