Non-Invasive Detection of Fetal Rhesus D Status: A Comparison between Polymerase Chain Reaction and Flow CytometryDi Simone N. · Lai M. · Rumi C. · Riccardi P. · D’Asta M. · Leone G. · Mancuso S. · Caruso A.
Departments of aObstetrics and Gynecology and bHematology, Catholic University of the Sacred Heart, Rome, Italy
Objective: A non-invasive prenatal determination of the fetal RhD status might be useful for the management of pregnancies in RhD-negative women whose partners are RhD positive. Methods: Maternal peripheral blood of 32 RhD-negative women (17–24 weeks of gestation) was collected, and circulating fetal cells were enriched by CD71 mini-magnetic activated cell sorting. The RhD status of the fetuses was assessed using multiparametric flow cytometry, and results were compared to those of reverse transcriptase (RT)-polymerase chain reaction (PCR), or PCR, which acted as control. Flow-cytometric study of fetal cells employed monoclonal antibodies directed against CD71, glycophorin A (GPA) and RhD antigens. Results: The median percentage of CD71- and RhD-positive cells was 0.83% (range 0.14–6.44%), and that of CD71 and GPA-positive cells was 10.07% (range 0.52–45.84%). Flow-cytometric analysis correlated with RT-PCR results of RNA obtained from whole maternal blood. In 1 case, an incorrect result was due to the failure of the amplification of the specific RhD band on RNA extracted from the CD71-positive fraction. In two instances, we observed false-positive results for RhD in PCR of DNA obtained from maternal plasma. Conclusion: Based on our results, flow-cytometric analysis might be proposed as a clinical tool for the non-invasive prenatal determination of the fetal RhD status independently of fetal gender.
Alessandro Caruso, MD
Department of Obstetrics and Gynecology, Catholic University of the Sacred Heart
Largo Gemelli 8
IT–00168 Rome (Italy)
Tel. +39 06 3015 6583, Fax +39 06 305 1160, E-Mail email@example.com
Supported by a grant from the Catholic University of the Sacred Heart (D3.2, year 2001).
Received: November 17, 2004
Accepted after revision: September 5, 2005
Number of Print Pages : 6
Number of Figures : 1, Number of Tables : 2, Number of References : 27
Fetal Diagnosis and Therapy (Clinical Advances and Basic Research)
Vol. 21, No. 5, Year 2006 (Cover Date: August 2006)
Journal Editor: Holzgreve, W. (Basel)
ISSN: 1015–3837 (print), 1421–9964 (Online)
For additional information: http://www.karger.com/FDT