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

产前诊断用母体外周血胎儿有核细胞富集技术的建立及临床应用

基本信息

批准号：
08457434
负责人：
KUBO Takeshi
金额：
$ 0.96万
依托单位：
University of Tsukuba
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
1996
资助国家：
日本
起止时间：
1996 至 1997
项目状态：
已结题

项目摘要

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.

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（有核红细胞）采用荧光激活细胞分选技术分离富集母血中的有核红细胞，并采用荧光原位杂交方法进行分析。研究结果显示了以下几个新的发现：结果表明：（1）流式细胞术结合特异性抗体具有分离富集孕妇外周血NRBC的潜力;（2）孕妇外周血中胎儿NRBC的频率随妊娠时间的延长呈下降趋势;（3）对已分选的男性胎儿孕妇外周血标本进行Y-荧光原位杂交（FISH）分析，结果显示：胎儿NRBC的频率随妊娠时间的延长呈下降趋势;（4）胎儿NRBC的频率随妊娠时间的延长呈下降趋势;（5）胎儿NRBC的频率随妊娠时间的延长呈下降趋势;（6）胎儿NRBC的频率随妊娠时间的延长呈下降趋势;（7）胎儿NRBC的频率随妊娠时间的延长呈下降趋势;（8）胎儿NRBC的频率随妊娠时间的延长呈下降趋势;（9）胎儿NRBC的频率随妊娠时间的延长呈下降趋势;（10）胎儿NRBC的频率随妊娠时间的延长呈下降趋势;（10）胎儿NRBC的频率随妊娠时间的延长呈下降趋势;（10）胎儿NRBC的频率随妊娠时间的延长呈下降趋势。特异性探针通过FISH检测分选的NRBC中的胎儿细胞。然而，95%的分选的NRBC是母体来源的。结果表明，流式细胞术对胎儿性别诊断的特异性为100%，敏感性为88%，阴性预测值为86%，阳性预测值为100%，可有效分离富集孕妇外周血中胎儿NRBC。这一新的诊断方法还需进一步完善，才能应用于临床遗传学诊断。