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

Di Simone, N., Lai, M., Rumi, C., Riccardi, P., D'asta, M., Leone, G., Mancuso, S., Caruso, A., Non-invasive detection of fetal Rhesus D Status: A comparison between Polymerase Chain Reaction and flow Cytometry, <<FETAL DIAGNOSIS AND THERAPY>>, 2006; (21(5)): 404-409. [doi:10.1159/000093880] [http://hdl.handle.net/10807/21252]

Non-invasive detection of fetal Rhesus D Status: A comparison between Polymerase Chain Reaction and flow Cytometry

Di Simone, Nicoletta;Lai, Marco;Rumi, Carlo;Riccardi, Patrizia;Leone, Giuseppe;Mancuso, Salvatore;Caruso, Alessandro
2006

Abstract

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
Inglese
Di Simone, N., Lai, M., Rumi, C., Riccardi, P., D'asta, M., Leone, G., Mancuso, S., Caruso, A., Non-invasive detection of fetal Rhesus D Status: A comparison between Polymerase Chain Reaction and flow Cytometry, <<FETAL DIAGNOSIS AND THERAPY>>, 2006; (21(5)): 404-409. [doi:10.1159/000093880] [http://hdl.handle.net/10807/21252]
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