Kleihauer抗酸染色法标记母血中的胎儿有核红细胞

以18例孕7~25周的孕妇外周血为材料, 经Percoll不连续密度梯度离心初步富集胎儿有核红细胞。然后用Kleihauer抗酸染色法进行标记, 结果阳性胎儿有核红细胞的胞浆呈深红色, 而母亲的有核红细胞胞浆无色。显微操作法获取单个胎儿有核红细胞, 经全基因组扩增后, 产物进行性别鉴定及STR连锁分析检测, 验证有核红细胞的来源, 并完成9例杜氏肌营养不良(Duchenne muscular dystrophy,DMD)的无创性产前基因诊断。应用Kleihauer抗酸染色法标记胎儿有核红细胞, 它是一种快速、简单、直接的化学染色方法, 更易于推广到临床应用。

Detection of fetal nucleated red blood cells in the maternal circula-tion by Kleihauer test

Maternal blood was obtained from 18 pregnant women at 7 to 25 weeks of gestation.After Percoll discontinu-ous density gradient centifugation,the fetal nucleated red blood cells(NRBCs) were stained with Kleihauer test.Positive fetal cells appeared with an intense red cytoplasmic staining while maternal cells with adult haemoglobin were colourless.Individual positive NRBC was collected by micromanipulator and whole genome amplification was then performed to de-termine sex and STR status.This allowed the simultaneous verification of the fetal origin of NRBC and prenatal diagnosis of genetic diseases.The non-invasive prenatal genetic diagnosis of 9 fetuses at high risk of Duchenne muscular dystrophy(DMD) was completed successfully.The Kleihauer test is a rapid,simple and direct chemical staining method to select fetal cells and can be applied in prenatal diagnosis.