PMID- 10419613 OWN - NLM STAT- MEDLINE DCOM- 19990819 LR - 20190905 IS - 0197-3851 (Print) IS - 0197-3851 (Linking) VI - 19 IP - 7 DP - 1999 Jul TI - Enrichment, identification and analysis of fetal cells from maternal blood: evaluation of a prenatal diagnosis system. PG - 648-52 AB - In this study we evaluated the performance of a system for the enrichment, identification and analysis of fetal cells in maternal peripheral blood. Blood samples were collected from women after chorionic villus sampling and enriched for the presence of nucleated erythrocytes using a three-step procedure, namely: (a) centrifugation to separate nucleated red blood cells (NRBCs) from the majority of red blood cells (RBCs) and white blood cells (WBCs); (b) selective lysis of the remaining maternal RBCs; (c) separating the NRBCs from the remaining WBCs in a three-layer density gradient. Fetal cells were identified by using a monoclonal antibody against the gamma-chain of fetal haemoglobin (anti-HbF) and a nuclear stain (DAPI). Additionally, to further increase the specificity of the identification, and to eliminate some of the undesired staining by maternal leukocytes, a fluorescent antibody (CD45) was added. The sex chromosome complement of the cells was determined by fluorescence in situ hybridization (FISH) with X and Y-specific probes and the results were compared with the karyotypes obtained after analysis of chorionic villi. Using the described method, in all cases where the woman was carrying a male fetus (n=18) at least one XY cell was found, while no male cells were found in women carrying a female fetus. However, in the majority of cases with a male fetus (n=11) female HbF positive cells were found indicating the presence of maternal nucleated erythrocytes. The study demonstrates that the combination of anti-HbF and CD45 is a useful, but not fully specific, marker for fetal NRBCs and that additional markers are needed. CI - Copyright 1999 John Wiley & Sons, Ltd. FAU - de Graaf, I M AU - de Graaf IM AD - Department of Clinical Genetics, Academic Medical Centre, Amsterdam, The Netherlands. I.M.deGraaf@AMC.UvA.Nl FAU - Jakobs, M E AU - Jakobs ME FAU - Leschot, N J AU - Leschot NJ FAU - Ravkin, I AU - Ravkin I FAU - Goldbard, S AU - Goldbard S FAU - Hoovers, J M AU - Hoovers JM LA - eng PT - Journal Article PT - Research Support, Non-U.S. Gov't PL - England TA - Prenat Diagn JT - Prenatal diagnosis JID - 8106540 RN - 0 (Antibodies, Monoclonal) RN - 9034-63-3 (Fetal Hemoglobin) RN - EC 3.1.3.48 (Leukocyte Common Antigens) SB - IM MH - Antibodies, Monoclonal MH - Cell Nucleus MH - Cell Separation/*methods MH - Centrifugation MH - Centrifugation, Density Gradient MH - *Erythrocytes/ultrastructure MH - Female MH - Fetal Blood/*cytology MH - Fetal Hemoglobin/analysis MH - Hemolysis MH - Humans MH - In Situ Hybridization, Fluorescence MH - Leukocyte Common Antigens/analysis MH - Male MH - Pregnancy MH - Prenatal Diagnosis/*methods MH - Sensitivity and Specificity MH - Sex Chromosomes EDAT- 1999/07/27 00:00 MHDA- 1999/07/27 00:01 CRDT- 1999/07/27 00:00 PHST- 1999/07/27 00:00 [pubmed] PHST- 1999/07/27 00:01 [medline] PHST- 1999/07/27 00:00 [entrez] AID - 10.1002/(SICI)1097-0223(199907)19:7<648::AID-PD600>3.0.CO;2-X [pii] AID - 10.1002/(sici)1097-0223(199907)19:7<648::aid-pd600>3.0.co;2-x [doi] PST - ppublish SO - Prenat Diagn. 1999 Jul;19(7):648-52. doi: 10.1002/(sici)1097-0223(199907)19:7<648::aid-pd600>3.0.co;2-x.