title: A one-step real-time PCR assay for rapid prenatal diagnosis of sickle cell disease and detection of maternal contamination.
abstract: BACKGROUND
  Mutations at the codon 6 of the beta-globin gene (hemoglobin [Hb] S and HbC) can be routinely identified by various methods and prenatal diagnosis has been available to affected families for several years. However, the presence of maternal cells in fetal samples constitutes a serious potential source of prenatal misdiagnosis and most methods currently used to detect maternal contamination are based on the analysis of highly polymorphic loci. In addition, these methods are labor intensive and time consuming and risk carry-over contamination.
  
  
  METHODS
  We describe here a one-step method for mutation detection that uses fluorescent hybridization probes with melting curve analysis for both simultaneously prenatal diagnosis of sickle cell disease and potential maternal contamination.
  
  
  RESULTS
  Retrospective and prospective prenatal diagnosis studies (conducted in 20 and 50 cases, respectively), using both the regular procedure and real-time PCR assay show perfect concordant results. We show in addition, that as little as 5% maternal contamination can be detected and that genotype determinations are unambiguous.
authors.affiliation: Laboratoire de Génétique Moléculaire, CHU Henri Mondor AP-HP, Créteil, France. costa@im3.inserm.fr
authors.forename: Catherine
authors.initials: C
authors.lastname: Costa
authors.forename: Serge
authors.initials: S
authors.lastname: Pissard
authors.forename: Emmanuelle
authors.initials: E
authors.lastname: Girodon
authors.forename: Danièle
authors.initials: D
authors.lastname: Huot
authors.forename: Michel
authors.initials: M
authors.lastname: Goossens
mesh_terms: D000483:Alleles; D000653:Amniotic Fluid / Q000166:cytology*; D000755:Anemia, Sickle Cell / Q000175:diagnosis* / Q000235:genetics; D015193:Chorionic Villi Sampling; D003062:Codon; D005260:Female; D005838:Genotype; D005914:Globins / Q000235:genetics*; D006801:Humans; D008297:Male; D009154:Mutation; D016133:Polymerase Chain Reaction; D011247:Pregnancy; D011296:Prenatal Diagnosis / Q000379:methods*; D011446:Prospective Studies; D012189:Retrospective Studies
chemical_list: D003062:Codon; D005914:Globins