基于纳米技术从孕妇外周血中分离胎儿有核红细胞并进行无创性产前诊断的可行性

发布时间：2018-07-09 22:00

本文选题：纳米基底片 + CD　；参考：《广东医学》2017年02期

【摘要】：目的建立一种基于纳米技术从孕妇外周血中分离胎儿有核红细胞并进行无创性产前诊断的方法。方法 Percoll 1.077单密度梯度离心法之后,选用抗体CD147修饰到以玻璃为载体的纳米羟基磷灰石基底上,对新生儿脐血中的胎儿有核红细胞进行捕获,利用胎儿特异性抗体HbF、红细胞表面特异性抗体CD71以及细胞核染色剂DAPI对捕获到的新生儿脐血中有核红细胞进行染色标记,进而实现分选。对正常不同孕周孕妇外周血、未孕健康妇女外周血及新生儿脐血中抗体CD147捕获到的有核红细胞进行染色标记分选。抽取10例行常规产检时发现高危因素的孕妇外周血2 m L,用抗体CD147捕获胎儿有核红细胞并用X、Y、18、21染色体探针进行荧光原位杂交。结果用HbF+CD71双标记染色法的胎儿有核红细胞阳性率与HbF单染和CD71单染标记的细胞阳性率相比差异有统计学意义(P0.05);HbF单染和CD71单染标记的阳性细胞率之间相比差异无统计学意义(P0.05);在孕16~20周时孕妇外周血中阳性细胞数目达到顶峰,之后随孕周增加而呈下降趋势。10例高危孕妇外周血标本均成功分离出胎儿有核红细胞,并经过FISH鉴定,本方法对男性胎儿诊断的灵敏度为85.71%,特异度为100.00%,对女性胎儿诊断的灵敏度为100.00%,特异度为85.71%,同时诊断出1例非整倍染色体胎儿,所有结果均经后期随访证实。结论用纳米材料为基底,抗体CD147捕获,HbF+CD71双染色标记法对孕妇外周血中的胎儿有核红细胞进行识别和富集,可提高胎儿有核红细胞的检出率,可为进一步产前诊断奠定基础,最佳时机应该在孕16~20周。
[Abstract]:Objective to establish a non-invasive prenatal diagnosis method for the separation of fetal nucleated red blood cells from maternal peripheral blood based on nanotechnology. Methods after percoll 1.077 single density gradient centrifugation, antibody CD147 was modified onto the nano-hydroxyapatite substrate with glass carrier to capture fetal nucleated red blood cells from umbilical cord blood of newborn. Fetal specific antibody HbF, erythrocyte surface specific antibody CD71 and nuclear staining agent DAPI were used to stain and label nuclear red blood cells in captured umbilical cord blood. The nucleated red blood cells captured by CD147 in the peripheral blood of normal pregnant women with different gestational weeks and those of healthy women without pregnancy and umbilical cord blood of newborns were stained and sorted. Ten maternal peripheral blood samples with high risk factors were collected from 10 routine birth tests. Fetal nucleated red blood cells were captured by antibody CD147 and fluorescence in situ hybridization was performed with the XMY 1821 chromosome probe. Results the positive rate of fetal nucleated red blood cells using HbF CD71 double labeling method was significantly different from that of HBF single staining and CD71 labeled cells (P0.05) there was no difference between the positive rates of HBF single staining and CD71 single staining positive cells. The number of positive cells in peripheral blood of pregnant women reached the peak at 1620 weeks of gestation (P0.05). The fetal nucleated erythrocytes were isolated from peripheral blood samples of 10 high-risk pregnant women and identified by fish. The sensitivity and specificity of this method for the diagnosis of male fetus were 85.71 and 100.000.The sensitivity and specificity for female fetus were 100.00and 85.71respectively, and a case of aneuploid chromosome fetus was diagnosed. All the results were confirmed by late follow-up. Conclusion using nanomaterials as the substrate, the detection rate of fetal nucleated red blood cells in maternal peripheral blood can be improved by double staining method of antibody CD147 capture HbF CD71, which can lay a foundation for further prenatal diagnosis. The best time to get pregnant is 16 to 20 weeks.
【作者单位】：承德市中心医院产科;
【分类号】：R714.5

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