Prenatal Diagnosis of Common Aneuploidies in Transcervical Samples Using Quantitative Fluorescent-PCR Analysis
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ORIGINAL RESEARCH ARTICLE
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Prenatal Diagnosis of Common Aneuploidies in Transcervical Samples Using Quantitative Fluorescent-PCR Analysis Cecilia Bussani, Riccardo Cioni, Alberto Mattei, Massimiliano Fambrini, Mauro Marchionni and Gianfranco Scarselli Department of Gynecology, Perinatology and Human Reproduction, University of Florence, Florence, Italy
Abstract
Aim: The aim of this study was to test the feasibility of diagnosing common fetal chromosomal aneuploidies using quantitative fluorescent (QF)-PCR on transcervical cell (TCC) samples collected in the first trimester of pregnancy by means of intrauterine lavage (IUL). Methods: A total of 181 TCC samples were retrieved from pregnant women between 5 and 12 weeks of gestation, immediately before elective termination of pregnancy, at which time corresponding placental tissue and maternal blood specimens were also obtained. Isolation of trophoblastic cells by micromanipulation was attempted in all TCC samples. Micromanipulated specimens were analyzed by multiplex QF-PCR, including short tandem repeats for the chromosomes X, Y, 21, 18, and 13. Results: The micromanipulation was successful in 152 of 181 cases (84.8%) where chorionic villous filaments and/or cell clumps of seeming trophoblastic origin could be isolated. All 152 samples were tested by QF-PCR analysis and peaks of paternal origin could be documented in all cases. Two cases of trisomy 21 and two cases of monosomy X0 were detected by means of QF-PCR assay, in accordance with the results obtained in corresponding placental samples. Conclusion: This study provides evidence that the use of multiplex QF-PCR amplification of selected microsatellites could be applied to micromanipulated TCC samples and in particular to IUL samples, which often contain trophoblastic cells, for the detection of chromosomal aneuploidies. The approach described in this study appears, therefore, a very promising tool toward non-invasive prenatal genetic diagnosis in the early stage of gestation.
Background Prenatal diagnosis of fetal genetic disorders started in the early 1970s and over the years has become increasingly common. The main methods for fetal-cell retrieval are represented by chorionic villus sampling (CVS) and amniocentesis, performed at 10–12 and 16–18 weeks gestation, respectively. Both these procedures are invasive and have been associated with a risk of pregnancy loss of 1%.[1] The possibility of performing prenatal genetic diagnosis by non- or minimally invasive techniques has been extensively studied. In particular, in recent years investigators have focused on the isolation and analysis of fetal cells and cell-free fetal DNA from maternal circulation and also on the retrieval of trophoblastic elements in the cervix and the uterine cavity.
Although important questions have been raised as to the biological significance of fetal cells and DNA in maternal blood, implications for prenatal genetic diagnosis are currently poor, owing to the overwhelming presenc
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