Bianchi D W, Shuber A P, DeMaria M A, Fougner A C, Klinger K W
Department of Pediatrics, New England Medical Center, Boston, MA.
Am J Obstet Gynecol. 1994 Oct;171(4):922-6. doi: 10.1016/s0002-9378(94)70059-1.
The detection of fetal aneuploidy and gene mutations by analysis of fetal cells in maternal blood has demonstrated the feasibility of noninvasive prenatal diagnosis. Fetal cells are rare in the maternal circulation; all current methods used for their isolation also yield maternal cells. We developed a method that permits a quantitative assessment of the relative numbers of fetal and maternal cells.
Samples from 40 pregnant women were flow sorted with different monoclonal antibodies. Deoxyribonucleic acid was subsequently purified from candidate fetal cells; polymerase chain reaction was performed with synthetic primers specific for sequences on chromosomes Y and 7.
The maximum number of fetal cells detected was 52 in 1080 maternal cells. Fetal cell purity ranged from 0.001% to 4.8%. Fetal cells were detected with antibodies to CD71, CD36, and glycophorin A.
Quantitative polymerase chain reaction enables the determination of the purity and yield of fetal cells remaining after isolation from maternal blood, facilitating rapid comparisons between different cell separation techniques.
通过分析母血中的胎儿细胞来检测胎儿非整倍体和基因突变，已证明了无创产前诊断的可行性。胎儿细胞在母血循环中很罕见；目前用于分离胎儿细胞的所有方法也会分离出母体细胞。我们开发了一种方法，可对胎儿和母体细胞的相对数量进行定量评估。
用不同的单克隆抗体对40名孕妇的样本进行流式分选。随后从候选胎儿细胞中纯化脱氧核糖核酸；使用针对Y染色体和7号染色体上序列的合成引物进行聚合酶链反应。
在1080个母体细胞中检测到的胎儿细胞最多为52个。胎儿细胞纯度范围为0.001%至4.8%。用抗CD71、CD36和血型糖蛋白A的抗体检测到了胎儿细胞。
定量聚合酶链反应能够确定从母血中分离后剩余的胎儿细胞的纯度和产量，便于快速比较不同的细胞分离技术。
