title: Reliability of polymerase chain reaction (PCR) analysis of single cells for preimplantation genetic diagnosis.
abstract: OBJECTIVE
  We investigated the reliability of polymerase chain reaction (PCR) genotype analyses performed on single cells for the purposes of preimplantation genetic analysis.
  
  
  METHODS
  We performed blind analysis of 130 single skin fibroblasts heterozygous for the delta-F508 mutation in the cystic fibrosis transmembrane regulator (CFTR) gene and 73 single skin fibroblasts from an individual heterozygous for the XbaI polymorphic site of the Factor VIII gene.
  
  
  RESULTS
  Amplification was successful for 116 cells and 52 cells respectively and in all but one case (a CFTR analysis) both alleles were amplified. The incidence of diagnostic error was 1 out of 203 analyses or 0.0043. We conclude that PCR is a reliable method for determining the genotype of single cells for the purposes of preimplantation genetic analysis.
authors.affiliation: Reproductive Genetics Institute, Illinois Masonic Medical Center, Chicago 60657.
authors.forename: C M
authors.initials: CM
authors.lastname: Strom
authors.forename: S
authors.initials: S
authors.lastname: Rechitsky
authors.forename: G
authors.initials: G
authors.lastname: Wolf
authors.forename: Y
authors.initials: Y
authors.lastname: Verlinsky
mesh_terms: D000483:Alleles; D000653:Amniotic Fluid / Q000166:cytology; D001483:Base Sequence; D001755:Blastocyst; D003550:Cystic Fibrosis / Q000175:diagnosis* / Q000196:embryology / Q000235:genetics; D019005:Cystic Fibrosis Transmembrane Conductance Regulator; D004247:DNA / Q000235:genetics*; D017931:DNA Primers; D015252:Deoxyribonucleases, Type II Site-Specific; D003951:Diagnostic Errors; D005169:Factor VIII / Q000235:genetics; D005315:Fetal Diseases / Q000175:diagnosis*; D005347:Fibroblasts / Q000737:chemistry*; D006580:Genetic Carrier Screening; D005838:Genotype; D006801:Humans; D008565:Membrane Proteins / Q000235:genetics; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D011296:Prenatal Diagnosis; D016037:Single-Blind Method; D012867:Skin / Q000166:cytology
chemical_list: C505032:CFTR protein, human; D017931:DNA Primers; D008565:Membrane Proteins; D019005:Cystic Fibrosis Transmembrane Conductance Regulator; D005169:Factor VIII; D004247:DNA; C025580:endodeoxyribonuclease XBAI; D015252:Deoxyribonucleases, Type II Site-Specific