OBJECTIVES: To establish the genotype of cultured cells from a cohort of amniotic fluid and chorionic villus samples, and compare this genotype with that obtained from uncultured material from the same sample, in order to assess the frequency and significance of maternal cell contamination of prenatal samples. METHODS: Quantitative fluorescence-polymerase chain reaction (QF-PCR) was carried out by amplification of microsatellite markers using fluorescence-labelled primers, followed by quantitative analysis of the allele peaks on a genetic analyser. A multiplex of 12 primer pairs for four loci on each of chromosomes 13, 18 and 21 was used. RESULTS: A total of 307 prenatal samples were tested. Of the 254 amniotic fluid samples, 39.8% had some degree of bloodstaining, ranging from 5% bloodstaining in the cell pellet to heavily bloodstained fluid. Uncultured samples were tested by QF-PCR analysis and the cultured cells were tested by both QF-PCR and karyotype analysis. Of the samples, 90.2% had the same single genotype on direct and cultured material. Two samples (0.65%) were mosaic for an aneuploidy cell line. A second genotype, interpreted as maternal cell contamination, was identified in direct and/or cultured preparations in 9.1% of samples, 17.8% of which were not bloodstained. Seven amniotic fluid samples (2.8%) showed maternal cell contamination in cultured material. CONCLUSIONS: For heavily bloodstained amniotic fluid samples, a maternal blood specimen may help interpret the results of rapid trisomy testing, followed by confirmation of the fetal origin of cultured cells. QF-PCR analysis has established a higher incidence of maternal cell contamination of cultured amniocytes than previous reports; the presence of MCC (maternal cell contamination) in cultured cells from samples with no bloodstaining underlines the need for karyotype analysis of more than one XX culture. Copyright (c) 2005 John Wiley & Sons, Ltd.
OBJECTIVES: To establish the genotype of cultured cells from a cohort of amniotic fluid and chorionic villus samples, and compare this genotype with that obtained from uncultured material from the same sample, in order to assess the frequency and significance of maternal cell contamination of prenatal samples. METHODS: Quantitative fluorescence-polymerase chain reaction (QF-PCR) was carried out by amplification of microsatellite markers using fluorescence-labelled primers, followed by quantitative analysis of the allele peaks on a genetic analyser. A multiplex of 12 primer pairs for four loci on each of chromosomes 13, 18 and 21 was used. RESULTS: A total of 307 prenatal samples were tested. Of the 254 amniotic fluid samples, 39.8% had some degree of bloodstaining, ranging from 5% bloodstaining in the cell pellet to heavily bloodstained fluid. Uncultured samples were tested by QF-PCR analysis and the cultured cells were tested by both QF-PCR and karyotype analysis. Of the samples, 90.2% had the same single genotype on direct and cultured material. Two samples (0.65%) were mosaic for an aneuploidy cell line. A second genotype, interpreted as maternal cell contamination, was identified in direct and/or cultured preparations in 9.1% of samples, 17.8% of which were not bloodstained. Seven amniotic fluid samples (2.8%) showed maternal cell contamination in cultured material. CONCLUSIONS: For heavily bloodstained amniotic fluid samples, a maternal blood specimen may help interpret the results of rapid trisomy testing, followed by confirmation of the fetal origin of cultured cells. QF-PCR analysis has established a higher incidence of maternal cell contamination of cultured amniocytes than previous reports; the presence of MCC (maternal cell contamination) in cultured cells from samples with no bloodstaining underlines the need for karyotype analysis of more than one XX culture. Copyright (c) 2005 John Wiley & Sons, Ltd.
Authors: Diane Van Opstal; Marjan Boter; Danielle de Jong; Cardi van den Berg; Hennie T Brüggenwirth; Hajo I J Wildschut; Annelies de Klein; Robert-Jan H Galjaard Journal: Eur J Hum Genet Date: 2008-09-10 Impact factor: 4.246
Authors: Kelly J Duffy; Jack Littrell; Adam Locke; Stephanie L Sherman; Michael Olivier Journal: Nucleic Acids Res Date: 2008-10-21 Impact factor: 16.971