title: Rapid detection of the deltaF508 mutation in single cells using DHPLC: implications for preimplantation genetic diagnosis.
abstract: OBJECTIVE
  Practice of preimplantation genetic diagnosis (PGD) requires efficient amplification and analysis techniques. We have tested Denaturing High Performance Liquid Chromatography (DHPLC) to screen the deltaF508 mutation in heterozygous single cells in order to assess its usefulness for PGD of cystic fibrosis.
  
  
  METHODS
  One hundred and two single lymphocytes--including N/N and N/deltaF508--were studied. F508 locus was amplified by nested PCR followed by the analysis of PCR products by DHPLC in non-denaturing conditions.
  
  
  RESULTS
  On the basis of PCR-amplified product analysis, total efficiency of amplification was 98.78% (101/102), and allele dropout (ADO) rate was 3.7% (3/81). For each sample, results were obtained in less than 4 min with high resolution.
  
  
  CONCLUSIONS
  DHPLC is a rapid and efficient technique to detect the deltaF508 mutation in single cells and is therefore appropriate for clinical application of preimplantation genetic diagnosis of cystic fibrosis.
authors.affiliation: Laboratoire de Génétique Moléculaire, Centre Hospitalo-Universitaire, Montpellier, France. girardet@igh.cnrs.fr
authors.forename: Anne
authors.initials: A
authors.lastname: Girardet
authors.forename: Philippe
authors.initials: P
authors.lastname: Cathala
authors.forename: Mireille
authors.initials: M
authors.lastname: Claustres
mesh_terms: D002851:Chromatography, High Pressure Liquid; D003550:Cystic Fibrosis / Q000175:diagnosis*; D019005:Cystic Fibrosis Transmembrane Conductance Regulator / Q000235:genetics*; D005260:Female; D006579:Heterozygote; D006720:Homozygote; D006801:Humans; D008214:Lymphocytes; D009154:Mutation; D016133:Polymerase Chain Reaction; D011247:Pregnancy; D019836:Preimplantation Diagnosis
chemical_list: C505032:CFTR protein, human; D019005:Cystic Fibrosis Transmembrane Conductance Regulator