title: Improving clinical preimplantation genetic diagnosis for cystic fibrosis by duplex PCR using two polymorphic markers or one polymorphic marker in combination with the detection of the DeltaF508 mutation.
abstract: Cystic fibrosis (CF) is an autosomal recessive disease characterized by obstruction and chronic infection of the respiratory tract and pancreatic insufficiency. The first preimplantation genetic diagnosis (PGD) for CF was carried out in 1992. At our centre the first cycle was performed in 1993. However, the number of known CF mutations is >1000, so developing mutation-specific PCR protocols for PGD is unfeasible. This is why a number of marker-based duplex PCRs were developed at the single cell level. A duplex PCR of a mutation and one or two microsatellites is not only a diagnostic tool, but it can also be used as a control for allele drop-out and contamination. During PGD, embryos obtained in vitro are analysed for the presence or absence of a particular genetic disease, after which only embryos shown to be free of this disease are returned to the mother. In total, 22 PGD cycles with duplex PCR (IVS8CA/IVS17BTA, DeltaF508/IVS8CA, DeltaF508/IVS17BTA and D7S486/D7S490) were carried out in 16 couples, which resulted in four ongoing pregnancies and one miscarriage.
authors.affiliation: Centre for Medical Genetics, University Hospital and Medical School, Dutch-speaking Brussels Free University, Brussels, Belgium. veerle.goossens@az.vub.ac.be
authors.forename: V
authors.initials: V
authors.lastname: Goossens
authors.forename: K
authors.initials: K
authors.lastname: Sermon
authors.forename: W
authors.initials: W
authors.lastname: Lissens
authors.forename: M
authors.initials: M
authors.lastname: De Rycke
authors.forename: B
authors.initials: B
authors.lastname: Saerens
authors.forename: A
authors.initials: A
authors.lastname: De Vos
authors.forename: P
authors.initials: P
authors.lastname: Henderix
authors.forename: H
authors.initials: H
authors.lastname: Van de Velde
authors.forename: P
authors.initials: P
authors.lastname: Platteau
authors.forename: A
authors.initials: A
authors.lastname: Van Steirteghem
authors.forename: P
authors.initials: P
authors.lastname: Devroey
authors.forename: I
authors.initials: I
authors.lastname: Liebaers
mesh_terms: D001706:Biopsy; D001757:Blastomeres / Q000378:metabolism; D003550:Cystic Fibrosis / Q000175:diagnosis* / Q000196:embryology / Q000235:genetics*; D005260:Female; D005819:Genetic Markers / Q000235:genetics; D005820:Genetic Testing / Q000379:methods*; D006579:Heterozygote; D006801:Humans; D008214:Lymphocytes / Q000166:cytology / Q000378:metabolism; D008297:Male; D009154:Mutation / Q000235:genetics*; D016133:Polymerase Chain Reaction / Q000379:methods*; D011110:Polymorphism, Genetic / Q000235:genetics*; D019836:Preimplantation Diagnosis / Q000379:methods*; D020554:Sperm Injections, Intracytoplasmic
chemical_list: D005819:Genetic Markers