title: Optimal polymerase chain reaction amplification for preimplantation diagnosis in cystic fibrosis (delta F508)
abstract: OBJECTIVE
  To evaluate direct polymerase chain reaction amplification of mutation on single embryo cells for the routine preimplantation diagnosis of cystic fibrosis.
  
  
  METHODS
  Direct polymerase chain reaction amplification of mutation was performed to identify the cystic fibrosis delta F508 mutation in human blood DNA, single lymphocytes, embryos, and embryo cells obtained by biopsy. Preimplantation diagnosis was performed for a couple who were heterozygous carriers of the delta F508 mutation.
  
  
  METHODS
  Laboratory for preimplantation diagnosis in a reproductive medicine unit.
  
  
  METHODS
  Correct diagnosis of homozygous normal, heterozygous, and homozygous abnormal DNA of the cystic fibrosis delta F508 mutation.
  
  
  RESULTS
  45 blood samples (18 homozygous normal, 17 heterozygous, and 10 homozygous abnormal) and 204 single lymphocytes from known sources showed 100% amplification and were diagnosed correctly. 17 human embryos and 52 normal nucleated embryo cells obtained by single cell embryo biopsy also showed 100% amplification. After a miscarriage of the initial pregnancy (diagnosed at preimplantation to be homozygous normal) in the heterozygous carrier couple, fetal tissue was confirmed to be homozygous normal.
  
  
  CONCLUSIONS
  Direct polymerase chain reaction amplification of mutation is a simple, fast, reliable test for the common cystic fibrosis mutation (delta F508) in blood DNA and single cells and should be applicable to routine programmes of general screening, maternal blood examination, and preimplantation diagnosis.
authors.affiliation: Department of Obstetrics and Gynaecology, University of Adelaide, Queen Elizabeth Hospital, Woodville, Australia.
authors.forename: K H
authors.initials: KH
authors.lastname: Cui
authors.forename: E A
authors.initials: EA
authors.lastname: Haan
authors.forename: L J
authors.initials: LJ
authors.lastname: Wang
authors.forename: C D
authors.initials: CD
authors.lastname: Matthews
mesh_terms: D001483:Base Sequence; D003550:Cystic Fibrosis / Q000175:diagnosis* / Q000235:genetics; D019005:Cystic Fibrosis Transmembrane Conductance Regulator; D004247:DNA / Q000235:genetics; D017931:DNA Primers; D003951:Diagnostic Errors; D047108:Embryonic Development; D005260:Female; D006579:Heterozygote; D006720:Homozygote; D006801:Humans; D008565:Membrane Proteins / Q000235:genetics*; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*; D011247:Pregnancy; D011296:Prenatal Diagnosis / Q000379:methods*; D017384:Sequence Deletion
chemical_list: C505032:CFTR protein, human; D017931:DNA Primers; D008565:Membrane Proteins; D019005:Cystic Fibrosis Transmembrane Conductance Regulator; D004247:DNA