Durrant L G, Martin W L, McDowall K M, Liu D T
Department of Clinical Oncology, University of Nottingham, City Hospital, UK.
Early Hum Dev. 1996 Dec 30;47 Suppl:S79-83. doi: 10.1016/s0378-3782(96)01827-0.
We present the results of a study of fetal cell isolation from the peripheral blood of 46 women in the first trimester of pregnancy. The trophoblasts were sorted with paramagnetic beads labelled with a novel monoclonal antibody 340 (Mab340) (Durrant et al., Prenat. Diagn., 14 (1994) 131). This was followed by triple density gradient enrichment to remove maternal lymphocytes and red blood cells. Nucleated red blood cells (NRBC) were sorted by incubation with ferromagnetic particles coated with Mab CD71, an antitransferrin receptor monoclonal antibody, and separation on a mini-MACS column. Sorted cells were sexed using nested PCR for the Y chromosome and the results compared with the karyotypic analysis of the CVS. The sensitivity in determining a male pregnancy with NRBC alone was 38% and with trophoblasts alone was 39%. Sorting for both cell types correctly predicted a male pregnancy in 10/18 or 56%. Of the 10 males correctly identified, 3 were diagnosed on NRBC alone, 3 on trophoblast alone and 4 with both cell types. As there are very few fetal cells in maternal blood, sorting for both will increase the yield and improve diagnosis. However the technique requires further development to improve sensitivity.
我们展示了一项从46名孕早期女性外周血中分离胎儿细胞的研究结果。用标记有新型单克隆抗体340（Mab340）的顺磁性微珠对滋养层细胞进行分选（Durrant等人，《产前诊断》，14(1994)131）。随后进行三重密度梯度富集以去除母体淋巴细胞和红细胞。通过与包被有抗转铁蛋白受体单克隆抗体Mab CD71的铁磁性颗粒孵育并在微型MACS柱上分离，对有核红细胞（NRBC）进行分选。使用针对Y染色体的巢式PCR对分选后的细胞进行性别鉴定，并将结果与绒毛取样（CVS）的核型分析进行比较。仅用NRBC确定男性妊娠的敏感性为38%，仅用滋养层细胞的敏感性为39%。对两种细胞类型进行分选可在10/18例（即56%）中正确预测男性妊娠。在正确识别的10例男性中，3例仅通过NRBC诊断，3例仅通过滋养层细胞诊断，4例通过两种细胞类型诊断。由于母体血液中的胎儿细胞非常少，对两种细胞进行分选将提高产量并改善诊断。然而，该技术需要进一步发展以提高敏感性。
