title: Multiple mutation analysis of the cystic fibrosis gene in single cells.
abstract: PGD is becoming an alternative to prenatal diagnosis. The combination of IVF techniques with the PCR technology allows for the detection of genetic abnormalities in first polar bodies from oocytes and blastomeres from cleavage-stage embryos. Dealing with a genetic disease with a heterogeneous spectrum of mutations like cystic fibrosis, one of the objectives of centres offering PGD is the application of simple and efficient protocols that allow for the detection of a wide range of mutations with a single procedure. In the present work, 29 normal loci and the 31 most frequent cystic fibrosis transmembrane conductance regulator (CFTR) mutations in Southern Europe could be detected at the same time in single cells applying a modified and improved primer extension preamplification-PCR. Two different Taq polymerases were tested in isolated buccal cells heterozygous for several mutations. The protocol that gave statistically significant better results was also successful in oocytes and their first polar bodies.
authors.affiliation: Unitat de Biologia Cel.lular i Genètica Mèdica, Facultat de Medicina, Universitat Autònoma de Barcelona, 08193 Bellaterra and Hospital Sant Joan de Deu, Barcelona, Spain. jsanchezg@servet.uab.es
authors.forename: Jorge Fernando
authors.initials: JF
authors.lastname: Sánchez-García
authors.forename: Jordi
authors.initials: J
authors.lastname: Benet
authors.forename: Cristina
authors.initials: C
authors.lastname: Gutiérrez-Mateo
authors.forename: José
authors.initials: J
authors.lastname: Luís Séculi
authors.forename: Eugènia
authors.initials: E
authors.lastname: Monrós
authors.forename: Joaquima
authors.initials: J
authors.lastname: Navarro
mesh_terms: D000483:Alleles; D003550:Cystic Fibrosis / Q000175:diagnosis*; D019005:Cystic Fibrosis Transmembrane Conductance Regulator / Q000235:genetics*; D004252:DNA Mutational Analysis / Q000379:methods*; D005260:Female; D006801:Humans; D009061:Mouth Mucosa / Q000166:cytology; D009154:Mutation; D011247:Pregnancy; D011296:Prenatal Diagnosis / Q000379:methods*
chemical_list: C505032:CFTR protein, human; D019005:Cystic Fibrosis Transmembrane Conductance Regulator