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孕妇外周血胎儿有核红细胞的分离与检测 |
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李智 夏铁安
【摘要】 目的 从孕妇血循环中分选出有核红细胞(NRBCs)，并证实其是胎儿来源的。 方法 选用红细胞特异单克隆抗体Glycophorin A(GPA)结合流式细胞术成功地从3例妊娠16～24周的孕妇血中分离出NRBCs，并用Y-染色体特异重复序列DNA探针(PY3.4)荧光原位杂交及聚合酶链反应技术检测Y-染色体特异序列(SRY) 结果 FISH结果，3例分选出的GPA阳性细胞中PY3.4杂交阳性率分别是31.4%、0、38%；其PCR结果，分别为阳性、阴性、阳性，并在引产后得到性别证实。结论 孕妇外周血中确实存在胎儿有核红细胞，采用非创伤性方法可以分离并检测它们。
【关键词】 胎儿有核红细胞 流式细胞术 荧光原位杂交
THE DETECTION AND ISOLATION OF FETAL NUCLEATED ERYTHROCYTES FROM MATERNAL BLOOD USING FLOW CYTOMETRY Li Zhi,Xia Tiean*.* Department of Clinic Laboratory,First School of Clinical Medicine,Beijing Medical University,Beijing 100034 P.R. China
【Abstract】 Objective To isolate fetal nucleated red blood cells (NRBCs) from maternal blood and to confirm its fetal origin.Methods Monoclonal antibody against the glycophorin A (GPA) was used to identify nucleated erythrocytes in the peripheral blood of pregnant women.Candidates fetal cells from 3 pregnancies at 16 to 24 weeks of gestation were isolated successfully.Sorted cells were also confirmed using Y-special probe (PY3.4) repeated sequences (SRY) in maternal blood from women carrying male fetuses were detected by polymerase chain reaction (PCR).Results The PY3.4 position hybridization rates were 31.5%,0 and 38% respectively in sorted GPA positive cells (GPA+) of 3 samples by FISH,and the results of GPA+ cells of the 3 cases by PCR were positive,negative and positive respectively,which were manifested by the gender after aborticide. Conclusion This study demonstrates that fetal NRBCs are present in the peripheral blood of pregnant women and it is possible to isolate and detect them.
【Key words】 Fetal nucleated erythrocytes Flow cytometry Fluorescence in situ hybridization
许多资料显示妊娠期间母体血循环中存在极少量胎儿有核细胞，是非创伤性产前基因诊断潜在的胎儿物质来源［1］。不同种类的胎儿细胞可以通过胎盘循环进入母体血液，它们是：合胞体滋养层细胞、淋巴细胞、粒细胞和有核红细胞。分离哪一类型的胎儿细胞用于产前诊断更为理想，目前仍不明确。有人认为胎儿有核红细胞(nucleated red blood cells，NRBCs)较佳。其原因为：(1)NRBCs不存在于正常人外周血中；(2)它是胎儿血液中的主要成分，数量较白细胞丰富得多，同时它含有胎儿基因的全部成分。对分选出的NRBCs[1] [2] 下一页
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