title: Preimplantation genetic diagnosis for achondroplasia: genetics and gynaecological limits and difficulties.
abstract: BACKGROUND
  We report the first attempts at preimplantation genetic diagnosis (PGD) and IVF and their accompanying difficulties for achondroplasia (ACH) patients.
  
  
  METHODS
  A PGD test was developed using fluorescent single cell PCR on lymphoblasts from patients and controls and from blastomeres from surplus IVF embryos. A specific digestion control based on the use of two fluorochromes was elaborated. Ovarian stimulation and oocyte retrieval were carried out using conventional protocols.
  
  
  RESULTS
  We performed 88 single cell tests for which amplification was obtained in 86 (97.7%) single lymphoblasts. Allele drop out (ADO) was observed in two out of 53 (3.7%) heterozygous lymphoblasts. If we combine the results from the blastomere testing from surplus embryos with those from PGD cycles and re-analysis after PGD, we obtained a PCR signal in 84% of cases of which 91% were correctly diagnosed at the G380 locus. A total of six cycles were performed resulting in three embryo transfers. We observed difficulties in ovarian stimulation and oocyte retrieval with affected female patients. No pregnancy was obtained.
  
  
  CONCLUSIONS
  A PGD test for ACH is now available at our centre but our initial practice raises questions on the feasibility of such a test, specially with affected female patients.
authors.affiliation: Service de Biologie de la Reproduction, SIHCUS-CMCO, CHU de Strasbourg, 19 rue Louis Pasteur, BP120, 67303 Schiltigheim cedex, France.
authors.forename: Céline
authors.initials: C
authors.lastname: Moutou
authors.forename: Catherine
authors.initials: C
authors.lastname: Rongieres
authors.forename: Karima
authors.initials: K
authors.lastname: Bettahar-Lebugle
authors.forename: Nathalie
authors.initials: N
authors.lastname: Gardes
authors.forename: Christophe
authors.initials: C
authors.lastname: Philippe
authors.forename: Stéphane
authors.initials: S
authors.lastname: Viville
mesh_terms: D000130:Achondroplasia / Q000175:diagnosis*; D000328:Adult; D001757:Blastomeres; D002461:Cell Line, Transformed; D005240:Feasibility Studies; D005260:Female; D005307:Fertilization in Vitro; D005456:Fluorescent Dyes; D006801:Humans; D016133:Polymerase Chain Reaction; D011247:Pregnancy; D019836:Preimplantation Diagnosis; D016896:Treatment Outcome
chemical_list: D005456:Fluorescent Dyes