title: Non-invasive prenatal diagnosis by isolation of both trophoblasts and fetal nucleated red blood cells from the peripheral blood of pregnant women.
abstract: OBJECTIVE
  To isolate fetal trophoblasts and nucleated red blood cells from the peripheral blood of pregnant women.
  
  
  METHODS
  Trophoblasts were isolated from whole blood of women in the first trimester of pregnancy by a specific monoclonal antibody, 340. Nucleated red blood cells were isolated by separating whole blood on a triple gradient, staining with ferromagnetic particles coated with an antitransferrin monoclonal antibody and separated on a mini magnetic activated cell sorting (MACS) column. Sorted cells were sexed using a nested polymerase chain reaction for a specific sequence on the Y chromosome and the sex was confirmed by karyotyping of chorionic villus samples.
  
  
  METHODS
  Patients between 10 and 14 weeks of pregnancy who were undergoing elective chorionic villus sampling for the detection of fetal aneuploidies.
  
  
  METHODS
  Fetal sex determined by polymerase chain reaction on fetal cells sorted from maternal blood.
  
  
  RESULTS
  When both trophoblasts and nucleated red blood cells were sorted, fetal sex was correctly predicted in 12/13 cases (92%), which included correct diagnosis of five of six male pregnancies. More importantly the two techniques were complementary, with only one male pregnancy being diagnosed on both trophoblasts and nucleated red blood cells, two being detected only with trophoblasts and two on nucleated red blood cells alone. No false positives (male signal from a female pregnancy) were diagnosed with either trophoblasts or nucleated red blood cells even with the highly sensitive nested polymerase chain reaction technique, which is very prone to contamination. This study also shows that it is possible to isolate both trophoblasts and nucleated red blood cells from the same sample of maternal blood.
  
  
  CONCLUSIONS
  Fetal cells can be isolated from maternal blood at around 10 weeks of pregnancy.
authors.affiliation: Department of Clinical Oncology, University of Nottingham, UK.
authors.forename: L
authors.initials: L
authors.lastname: Durrant
authors.forename: K
authors.initials: K
authors.lastname: McDowall
authors.forename: R
authors.initials: R
authors.lastname: Holmes
authors.forename: D
authors.initials: D
authors.lastname: Liu
mesh_terms: D000782:Aneuploidy; D002469:Cell Separation; D004912:Erythrocytes; D005189:False Positive Reactions; D005260:Female; D005312:Fetal Blood / Q000737:chemistry; D006801:Humans; D008297:Male; D016133:Polymerase Chain Reaction; D011247:Pregnancy; D011261:Pregnancy Trimester, First; D011287:Prejudice; D011296:Prenatal Diagnosis / Q000379:methods*; D012680:Sensitivity and Specificity; D014327:Trophoblasts / Q000473:pathology*