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目的 :研究磁激活细胞分离富集孕妇外周血胎儿有核红细胞进行产前诊断的可行性。方法 :2 1名孕 13 2 9周孕妇及 5名未妊娠妇女外周血 4 6ml,经单密度梯度离心和激活细胞分离 ,收集阳性组分抽提DNA ,经PCR扩增预测胎儿性别 ,并以出生后婴儿性别验证预测性别的准确性。结果 :以Y染色体特异引物介导的PCR扩增Y染色体特异片段检出的最低限量为 3 4个男性细胞或 3 4个男性细胞 / 3 5× 10 4 女性细胞。 2 1名孕妇分娩 13名男婴、8名女婴。男性DNA在 12名孕男胎孕妇外周血有核红细胞中被检出 ,在 1名孕男胎孕妇中未被检出 ;而在 8名孕女胎孕妇外周血有核红细胞及 5名未孕妇女外周血中均未检出男性DNA。预测胎儿性别的准确度为 95 2 3% ,假阴性率为 4 76 %。结论 :磁激活细胞分离富集孕妇外周血胎儿有核红细胞是一简单、快速、价廉的有效方法 ,被分离细胞DNA经PCR扩增后可用于胎儿性别的非侵入性产前诊断。
OBJECTIVE: To investigate the feasibility of prenatal diagnosis of fetal nucleated red blood cells (PBMCs) enriched by magnetic activated cells in pregnant women. Methods: Twenty-four pregnant women, 1329 pregnant women and 5 nonpregnant pregnant women were divided into two groups. The positive cells were extracted by single density gradient centrifugation and activated cells. The DNA was extracted and the fetal sex was predicted by PCR. Postnatal baby sex verification predicts gender accuracy. Results: A minimum of 34 male cells or 34 male cells / 35 × 10 4 female cells were detected by Y chromosome specific primers. 21 pregnant women gave birth to 13 boys and 8 girls. Male DNA was detected in nucleated red blood cells of peripheral blood of 12 pregnant women and pregnant women, but not detected in 1 pregnant pregnant women; while in 8 pregnant women pregnant women, peripheral blood red blood cells and 5 non-pregnant women No female DNA was detected in female peripheral blood. The accuracy of predicting fetal sex was 95 2 3% and the false negative rate was 4 76%. Conclusion: It is a simple, rapid and inexpensive method to separate and enrich fetal nucleated red blood cells in peripheral blood of pregnant women by magnetically activated cells. The isolated DNA can be used for noninvasive prenatal diagnosis of fetal sex by PCR amplification.