The establishment and clinnical application of a enrichment technique of fetal nucleated cells from maternal peripheral blood for prenatal diagnosis

本研究旨在建立一种利用孕妇外周血中胎儿有核细胞进行产前诊断的新方法。NRBC(有核红血球)。采用荧光激活细胞分选(FACS)技术分离和浓缩母血中的有核红细胞(NRBC)，并用FISH(荧光原位杂交)方法进行分析。研究结果表明：1.证实了结合特异性抗体的流式细胞仪具有从孕妇外周血中分离和浓缩NRBCs的潜力。2.孕妇外周血中胎儿NRBCs的频率随着妊娠的进展有下降的趋势。3.对有男性胎儿的孕妇的分选样本进行Y-特异性探针FISH分析。FISH检测分离的NRBCs中的胎儿细胞。然而，95%的NRBCs起源于母体。胎儿性别诊断的特异度为100%，灵敏度为88%，阴性预测值为86%，阳性预测值为100%，FACS具有从孕妇外周血中分离和浓缩胎儿NRBCs的有效潜力。这一新的诊断方法还需要进一步完善，以用于临床基因诊断。

The purpose of this study was to establish a new prenatal diagnostic method using fetal nucleated cells in maternal peripheral blood. The NRBCs (nucleated red blood. The NRBCs (nucleated red cells) in maternal blood were separated and enriched by FACS (fluorescence activated cell sorting) technique and were analyzed using FISH (fluorescence in situ hybridization) method. The results of the study showed several new findings as follows ;1.It was confirmed that FACS with a combination of specific antibodies had the potential to separate and enrich NRBCs from peripheral blood samples of pregnant women.2.The frequency of fetal NRBCs in maternal peripheral blood had a tendency to decrease as the pregnancies advanced.3.The sorted samples from pregnant women who had male fetuses were analyzed by FISH with Y-specific probe. The fetal cells in the sorted NRBCs were determined by FISH.95% of the sorted NRBCs were, however, maternal in origin.4. In this series, the specificity of the fetal sex diagnosis was 100%, the sensitivity was 88%, the negative predictive value was 86%, and the positive predictive value was 100%.FACS has the effective potential to separate and enrich fetal NRBCs from maternal peripheral blood. Further improvement of this new diagnostic method is needed to use in clinical genetic diagnosis.