母血中胎儿有核红细胞用于无创产前诊断的研究  

Screening for chromosomal aneuploidy using fetal nucleated red blood cells from maternal blood by fluorescence in situ hybridization

作　　者：刘霞[1] 徐营[2] 宋勤浩[1] 沈华祥[1] 金玉霞[1] 李素萍[1] 苗正友[1] 

机构地区：[1]嘉兴学院附属妇幼保健院产前诊断中心,314050 [2]嘉兴学院医学院组胚教研室

出　　处：《浙江医学》2012年第8期590-592,602,共4页Zhejiang Medical Journal

基　　金：浙江省科技厅面上项目（2009C33038）;浙江省卫生厅A类基金（2008A154）

摘　　要：目的 用分离到的胎儿有核红细胞,评价应用荧光原位杂交技术筛查染色体非整倍性的临床应用价值.方法 43例（孕周17～21＋4周）拟行产前诊断的孕妇外周血20ml,用双密度梯度离心和双抗体磁性活化细胞分选（magnetic activated cell sorting,MACS）方法富集孕妇外周血中胎儿有核红细胞,其中10例患者富集的细胞进行吉姆萨-瑞氏染色,10例进行抗人CD235a和CD71免疫荧光抗体染色,行细胞鉴定和计数.23例富集后的细胞用荧光染色体原位杂交（fluorescence chromosomal in situ hybridization,FISH）探针进行非整倍体分析,并与羊水培养后核型分析结果进行比较.结果 所有标本均分离出有核红细胞,并经组织学染色、免疫荧光特异抗体和FISH鉴定.23例FISH诊断中,未发现非整倍体异常;14例男性胎儿有13例发现Y荧光信号,1例未发现,女性胎儿中均发现2个X荧光信号.结论 荧光原位杂交技术可对经富集的孕妇外周血中胎儿细胞进行非整倍体诊断.Objective To evaluate screening for chromosomal aneuploidy using fetal nucleated red blood cells from ma- ternal peripheral blood by fluorescence in situ hybridization. Methods 20 ml maternal peripheral blood samples were collected from 43 cases with 17 ~ 21 weeks of gestation who required prenatal diagnosis. The fetal nucleated red blood cells were enriched by double-density gradient centrifugation and double-antibody magnetic activated cell sorting. Giemsa-Wright＇s staining was undertaken in 10 samples of enriched fetal nucleated red blood cells and anti-CD236a and CD71 immunofluorescence antibody staining was undertaken in another 10 samples for cell identification and counting. The chromosome aneuploidy analysis was performed in remaining 23 samples of enriched fetal nucleated red cells by fluorescence in situ hybridization and the results were compared to karyotype analysis of those from cultured amniotic fluid. Results The nucleated red cells from maternal peripheral blood were separated and identified by cytological and immunofluorescence staining and FISH technique. No abnormality of a- neuploidy was detected in 23 samples diagnosed by FISH. For gender identification, Y fluorescent signal was detected in 13 out of 14 male fetuses; and two X fluorescence signal was detected in all female fetuses. Conclusion The chromosomal aneuploidy can be diagnosed by FISH technology in fetal nucleated red blood cells from maternal peripheral blood samples.

关 键 词：荧光原位杂交 有核红细胞 无创 产前诊断 

分 类 号：R714.5[医药卫生—妇产科学]