Prenatal Diagnosis of Ingenited Metabolic Disorders Using Maternal Blood

母血产前诊断是一种对母胎无创的诊断方法。怀孕3-4个月的母亲血液中含有胎儿滋养细胞，1 - 2-3万个母细胞。这些有核红细胞表现出糖蛋白A、CD-71、CD36、Y特异性探针和血红蛋白f等表面抗原，利用这些标记，我们尝试用磁珠法、凝集素法和荧光细胞仪方法分离胎儿细胞。我们检测了胎儿细胞，并将其应用于唐氏综合征、非整倍体综合征和遗传病（戈谢病、法布里病）等染色体畸变的DNA诊断。(1)母血胎儿细胞分离方法：在征得知情同意的情况下，从20 ml孕母中分离出有核细胞。采用凝集素柱、磁珠柱、荧光细胞法分离胎儿有核细胞。此外，利用D18S474微卫星标记，我们可以从母体血液中找到男性胎儿细胞。在此基础上，利用cDNA微阵列技术，对mNA在胎儿单细胞中的表达进行了研究。结果表明我们需要更多的细胞来证明。(2)染色体疾病的DNA诊断胎儿细胞我们试图证明给DNA诊断唐氏综合征和非整倍体综合征的母亲血液。为了对这些疾病进行诊断，我们首先试图确定非整倍体综合征的发病机制。一些数据表明，这种疾病是由纺锤体组装检查点的基因缺陷引起的。(3)胎儿细胞DNA诊断遗传病：我们试图确定日本戈谢病和法布里病患者的DNA诊断。我们确定日本戈谢病的常见突变如下：L444P， 32%, f213i, 17%, D409H, 5.2%, ecNcl, 4.2%。另一方面，对日本法布里病患者的几个突变进行了跟踪；L 16 H、A37Vm、W209X、342Q、IVS1-1、ivs2 -2、ivs2 -2、IVS6+1等。这些结果可用于产前血检诊断。少

Prenatal diagnosis by maternal blood is non-invasive manner for mother and fetus. Blood from 3-4 months pregnant mother contain fetal trophoblasts, 1 to 2-30, 000 mother cells. These nucleated erythrocytes exhibit surface antigens such as glycophorin A, CD-71, CD36, Y specific probe and hemoglobin F. Using these markers, we tried to isolate fetal fells using magnetic beads method, lectin method and fluorocytometer methods. We have detected fetal cells and applied to the DNA diagnosis of chromosomal aberrations such as Down syndrome, Aneuploidy syndrome and genetic diseases (Gaucher, Fabry disease)(1) Isolation method of fetal cells from maternal blood :Fctal nucleated cells were isolated from 20 ml pregnant mothers which we obtained informed consent. Using lectin column, magnetic beads column, and fluorocytometry methods, we could isolated fetal nucleated cells. Furthermore, using D18S474 microsatelite marker, we could demonstrated male fetal cells from maternal blood. Furthermore, usi … More ng cDNA micro array method, we tried to study mNA expression in fetal single cells. The results suggest we need more cells to demonstrate.(2) Chromosomal disorders for DNA diagnosis by fetal cells We attempt to demonstrate to give a DNA diagnosis of Down syndrome and aneuploidy syndrome by maternal blood. In order to give a diagnosis these disorders, first we attempted to determine pathogenesis of Aneuploidy syndrome. Several data suggest that This disorder is caused by gene defect in spindle assembly checkpoints.(3) Genetic disorders by DNA diagnosis by fetal cells :We attempted to determine DNA diagnosis in Japanese patients with Gaucher and Fabry disease. We determined the common mutations in Japanese Gaucher disease as follows : L444P, 32%, F2 1 3 I, 17%, D409H, 5.2%, ecNcl, 4.2%. In other hand, Several mutations in Fabry disease from Japanese patients were follows ; L 1 6 H, A37Vm W209X, 342Q, IVS1-1, IVS5-2, IVS5-2, IVS6+1 and etc. These results could be applied for prenatal diagnosis by maternal blood. Less