Nagy Gyula Richárd, Bán Zoltán, Sipos Ferenc, Beke Artúr, Papp Csaba, Papp Zoltán
Department of Obstetrics and Gynecology, Semmelweis University, Faculty of Medicine, Budapest, Hungary.
Prenat Diagn. 2005 May;25(5):398-402. doi: 10.1002/pd.1167.
The isolation and analysis of fetal cells in maternal blood during pregnancy is under investigation as a means of noninvasive prenatal diagnosis. The aim of our study was to detect fetal gender from maternal peripherial blood samples during pregnancy with the detection and analysis of epsilon-haemoglobin-chain positive fetal nucleated red blood cells (NRBCs) collected by a micromanipulator. Here we report our first results.
We obtained maternal blood from 14 singleton pregnancies. After a double density gradient separation, magnetic activated cell sorting was performed by positive selection for nucleated red blood cells with anti-CD71. With the help of this enrichment step, followed by immunophenotyping with an anti-haemoglobin-epsilon monoclonal antibody, the isolation of the epsilon haemoglobin chain positive cells with micromanipulation could be done. We performed single cell fluorescent PCR analysis of these cells; we used primers for the amelogenin gene to detect fetal gender. We compared our findings with the results of amniocentesis.
Fetal gender was successfully determined in 11 out of 14 cases; among them, in 2 cases with Klinefelter syndrome (47,XXY).
The results of our study suggest that micromanipulation and QF-PCR analysis of anti-haemoglobin-epsilon fluorescent antibody stained fetal cells from maternal blood can be useful in prenatal diagnosis to detect fetal gender and promising to be improved to detect chromosomal abnormalities.
孕期母血中胎儿细胞的分离与分析作为一种非侵入性产前诊断方法正在研究中。我们研究的目的是通过用显微操作器收集并分析ε-血红蛋白链阳性的胎儿有核红细胞（NRBCs），从孕期母体外周血样本中检测胎儿性别。在此我们报告我们的初步结果。
我们从14例单胎妊娠孕妇获取母血。经过双重密度梯度分离后，通过用抗CD71对有核红细胞进行阳性选择来进行磁性激活细胞分选。借助这一富集步骤，随后用抗ε-血红蛋白单克隆抗体进行免疫表型分析，就可以通过显微操作分离出ε-血红蛋白链阳性细胞。我们对这些细胞进行单细胞荧光PCR分析；我们使用牙釉蛋白基因引物来检测胎儿性别。我们将我们的发现与羊膜穿刺术结果进行比较。
14例中有11例成功确定了胎儿性别；其中，2例为克兰费尔特综合征（47,XXY）。
我们的研究结果表明，对母血中抗ε-血红蛋白荧光抗体染色的胎儿细胞进行显微操作和QF-PCR分析可用于产前诊断以检测胎儿性别，并且有望改进用于检测染色体异常。
