免疫纳米磁珠分离孕妇外周血中胎儿有核红细胞的有效性研究OA
Efficacy of immunomagnetic nanobead-based isolation of fetal nucleated red blood cells from maternal peripher-al blood
目的 探索特异抗体标记纳米磁珠分离孕妇血中胎儿有核红细胞的有效性.方法 密度梯度离心法结合免疫纳米磁珠分离脐血中胎儿有核红细胞,流式细胞术分析目标细胞纯度并用免疫荧光法鉴定.对孕妇和健康未孕女性外周血分离后细胞进行免疫荧光法检测.对男胎脐血混合未孕女性外周血模拟样本、染色体正常胎儿和非整倍体胎儿母体外周血标本分离后细胞行荧光原位杂交(fluorescence in situ hybrid-ization,FISH)实验.结果 流式细胞术发现阳性和阴性细胞中特异性抗体荧光双阳性的比例分别为(90.16±5.77)％、(7.12±4.45)％.在阳性细胞中3种抗体荧光共表达,阴性细胞中未发现.来自孕妇外周血的阳性细胞三种抗体荧光共同表达,对照组中未发现.FISH在所有模拟样本分离后细胞中检出Y染色体,在5例正常胎儿母体外周血分离后细胞中有2例检出Y染色体,在4例明确非整倍体胎儿母体外周血分离后细胞中检出2例21-三体、1例13-三体和1例18-三体,与羊水核型分析结果一致.结论 免疫纳米磁珠可有效分离孕妇血中的胎儿有核红细胞,可用于胎儿非整倍体的检测.
Objective To evaluate the efficacy of isolating fetal nucleated red blood cells(FNRBCs)from mater-nal peripheral blood using antibody-labeled immunomagnetic nanobeads.Methods Fetal nucleated red blood cells were isolated from umbilical cord blood using density gradient centrifugation combined with immunomagnetic nanobead sep-aration.The purity of target cells was assessed by flow cytometry and further verified by immunofluorescence staining.Pe-ripheral blood samples from pregnant women and healthy non-pregnant controls were processed and analyzed using immu-nofluorescence.Simulated samples were prepared by mixing umbilical cord blood from male fetuses with peripheral blood from non-pregnant women.In addition,maternal peripheral blood samples from pregnancies with chromosomally normal fetuses and fetuses with aneuploidy were analyzed.Fluorescence in situ hybridization(FISH)was performed on isolated cells to evaluate chromosomal characteristics.Results Flow cytometry revealed that the proportions of double-positive cells for specific antibody fluorescence were(90.16±5.77)％in the positive fraction and(7.12±4.45)％in the nega-tive fraction.Triple co-expression of antibody signals was observed in positive cells but not in negative cells.Cells isola-ted from maternal peripheral blood of pregnant women showed co-expression of all three antibody markers,whereas no such expression was observed in the control group.FISH analysis detected Y chromosomes in all simulated samples.A-mong five maternal peripheral blood samples from pregnancies with chromosomally normal fetuses,Y chromosomes were detected in two cases.Among four samples from pregnancies with confirmed fetal aneuploidy,two cases of trisomy 21,one case of trisomy 13,and one case of trisomy 18 were identified,consistent with amniotic fluid karyotype results.Conclu-sion Immunomagnetic nanobeads can effectively isolate fetal nucleated red blood cells from maternal peripheral blood and may serve as a promising approach for noninvasive detection of fetal aneuploidy.
杨兴坤;周雅思;李炜辉;杜佳月;陈玥;夏承来
广东医科大学附属佛山妇女儿童医院产前诊断中心(广东佛山 528000)佛山普利盛生物科技有限公司(广东佛山 528000)广东医科大学附属佛山妇女儿童医院产前诊断中心(广东佛山 528000)广东医科大学附属佛山妇女儿童医院产前诊断中心(广东佛山 528000)||龙岗区人民医院产科(广东 深圳 518100)南方医科大学第二临床学院(广东 广州 510075)广东医科大学附属佛山妇女儿童医院胎儿医学研究所(广东佛山 528000)
医药卫生
免疫标记纳米磁珠胎儿有核红细胞无创性产前诊断出生缺陷防控
immunomagnetic nanobeadsfetal nucleated red blood cellsnoninvasive prenatal diagnosisbirth defect prevention
《广东医学》 2026 (6)
845-851,7
国家自然科学基金面上项目(82171713)
评论