孕妇外周血中胎儿细胞及胎儿DNA的检测  

Detection of fetal cells and DNA from peripheral blood of pregnant women.

作　　者：陈汉平[1] 贺桂芳[1] 沈怡[1] 张汉凤[1] 陶德定[1] 马庭元[1] 

机构地区：[1]华中科技大学同济医学院附属同济医院妇产科,武汉430030

出　　处：《中国实用妇科与产科杂志》2002年第10期596-598,共3页Chinese Journal of Practical Gynecology and Obstetrics

基　　金：卫生部科学基金资助 (基金编号 :96-2 -112 );湖北省自然科学基金资助 (基金编号 96J0 68)

摘　　要：目的　建立从孕妇外周血中分离有核红细胞 (NRBC)及DNA的可靠方法 ,并鉴定其来源。方法　对88例孕妇外周血分别通过密度梯度离心法 ,流式细胞术 (FCM )和荧光激活细胞分离技术 (FACS)分离NRBCs。应用套式PCR技术对 6 5例孕妇血浆DNA进行正常男性SRY基因检测。结果　①NRBCs:2 7例样品经密度梯度离心 ,其中 14例分选到 1～ 10个NRBCs。FACS技术对 6 1例样品进行转铁蛋白受体阳性细胞 (CD71+ )分选 ,所有样品均有CD71+ 细胞 ,其浓度为 (0 35± 0 2 5 )× 10 -2 。②胎儿DNA :4 6例怀男胎孕妇血浆DNA中SRY基因检测出率为 6 5 2 2 % (30 / 4 6 ) ,怀女胎的 19例样品SRY基因未检出率为 94 74 % (18/ 19)。结论　从孕妇外周血中分选胎儿NRBCs和血浆中胎儿DNA的方法已取得较大进展。Objective To search for a more reliable method of sorting fetal nucleated red blood cells(NRBCs)and DNA from maternal peripheral blood and to indentify origin of NRBCs and DNA.Methods NRBCs were isolated from peripheral blood of 88 pregnant women by density gradient centrifugation,flow cytometry(FCM)and fluorescence activated cell sorter(FACS)respectively.Nested primer polymerase chain reaction was used to detect normal male SRY gene from blood plasma DNA of 65 pregnant women.Results ①Fetal NRBCs:NRBCs were found in 14 of 27 maternal samples by density gradient centrifugation.The number of cells was from one to ten.Using FACS,CD71\++ cells among 61 samples were totally expressed,the frequency was (0 35±0 25)×10 -2 .②Fetal DNA:The detecting rate of the SRY gene in blood plasma DNA from 46 women carrying male fetuses was 65\^22%(30/46).Non detecting rate for 19 women carrying female tetuses was 94\^74%(18/19).Conclusion The methods of sorting fetal NRBCs and DNA have already made great progresses.The method for sorting fetal NRBCs and plasma DNA from maternal peripheral blood to diagnose genetic diseases seems to be one of the best non invasive prenatal diagnostic methods.

关 键 词：胎儿有核红细胞 胎儿DNA 产前诊断 

分 类 号：R714.5[医药卫生—妇产科学]