title: [Preimplantation diagnosis of delta F508 mutation of mucoviscidosis in transgenic mice].
abstract: OBJECTIVE
  The aim of our study was to master preimplantation diagnosis in transgenic mice.
  
  
  METHODS
  Stage 4 or 8 cells preembryos were collected from C57BL6xCBAF1 mice, three days after mating with transgenic C57BL6xSJLF1 mice homozygous for the human gene delta F508 mutation of cystic fibrosis. A single blastomere was sampled by microbiopsy and the preembryos transferred in Swiss mice foster mothers. Molecular biology for delta F508 mutation on a single blastomere was performed with a double PCR technique on a Perkin Elmer Cetus 9600.
  
  
  RESULTS
  The success rate of biopsy on the 122 preembryos with 4 cells was 90.2%, and 93.8% on the 128 prembryos with 8 cells. The rate of in vitro hatching was 84.6% after biopsy on 52 preembryos with 4 cells (92.2% for controls without biopsy, p > 0.05), and 90.0% for 50 preembryos with 8 cells (94.4% for controls without biopsy, p > 0.05). The rate of birth after biopsy and transfer of 42 preembryos with 4 cells was 66.7% (72.5% for controls transferred without biopsy, p > 0.05) and 70.3% for 37 preembryos with 8 cells (71.9% for controls, p > 0.05). No difference was evidenced for births weight or organ weight at 3 weeks between mice born from biopsied embryos and controls. Thirty-two double PCR were performed for the diagnosis of the cystic fibrosis delta F508 mutation, 20 on a single blastomere obtained by microbiopsy and 12 for various negative controls, with 100% specificity and 100% sensitivity.
  
  
  CONCLUSIONS
  Microbiopsy of preembryos with 4 cells or 8 cells does not alter their viability or further development. The double PCR technique on a single blastomere for the cystic fibrosis delta F508 mutation is reliable with our current methods of investigation.
authors.affiliation: Service de Gynécologie et Obstétrique, Hôpital Saint-Antoine, Paris.
authors.forename: Y Z
authors.initials: YZ
authors.lastname: Ji
authors.forename: J C
authors.initials: JC
authors.lastname: Nicolas
authors.forename: V
authors.initials: V
authors.lastname: Marechal
authors.forename: N
authors.initials: N
authors.lastname: Desire
authors.forename: A
authors.initials: A
authors.lastname: Pavirani
authors.forename: J
authors.initials: J
authors.lastname: Milliez
mesh_terms: D000818:Animals; D001707:Biopsy, Needle; D003550:Cystic Fibrosis / Q000175:diagnosis* / Q000235:genetics; D047108:Embryonic Development; D005260:Female; D008722:Methods; D051379:Mice; D008810:Mice, Inbred C57BL; D008808:Mice, Inbred CBA; D008822:Mice, Transgenic; D008967:Molecular Biology; D009154:Mutation; D016133:Polymerase Chain Reaction; D011247:Pregnancy; D011270:Pregnancy, Animal; D013997:Time Factors
vernacular_title: Diagnostic préimplantatoire de la mutation delta F508 de la mucoviscidose chez la souris transgénique.