Reliability of polymerase chain reaction (PCR) analysis of single cells for preimplantation genetic diagnosis

J Assist Reprod Genet. 1994 Feb;11(2):55-62. doi: 10.1007/BF02215989.

Abstract

Purpose: 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.

MeSH terms

  • Alleles
  • Amniotic Fluid / cytology
  • Base Sequence
  • Blastocyst*
  • Cystic Fibrosis / diagnosis*
  • Cystic Fibrosis / embryology
  • Cystic Fibrosis / genetics
  • Cystic Fibrosis Transmembrane Conductance Regulator
  • DNA / genetics*
  • DNA Primers
  • Deoxyribonucleases, Type II Site-Specific
  • Diagnostic Errors
  • Factor VIII / genetics
  • Fetal Diseases / diagnosis*
  • Fibroblasts / chemistry*
  • Genetic Carrier Screening
  • Genotype
  • Humans
  • Membrane Proteins / genetics
  • Molecular Sequence Data
  • Polymerase Chain Reaction*
  • Prenatal Diagnosis*
  • Single-Blind Method
  • Skin / cytology

Substances

  • CFTR protein, human
  • DNA Primers
  • Membrane Proteins
  • Cystic Fibrosis Transmembrane Conductance Regulator
  • Factor VIII
  • DNA
  • endodeoxyribonuclease XBAI
  • Deoxyribonucleases, Type II Site-Specific