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目的 探讨获取母血中纯化胎儿细胞的有效方法。方法 对 6 0名孕龄 7～ 40周、年龄 2 1～ 30岁的孕妇外周血进行不连续密度梯度离心 ,将分离后的细胞进行制片 ,显微镜下行显微操作分离胎儿有核红细胞 ,进行 Y染色体特异性DYZ1基因的聚合酶链反应 (PCR)。结果 (1)早孕期母血中胎儿 NRBC检出率为 2 0 % ,中孕期母血中胎儿 NRBC检出率为 71.4% ,晚孕期母血中胎儿 NRBC检出率为 6 .6 5 % ,三者之间有显著性差异 (P <0 .0 5 ) ;(2 )用显微操作技术获得的胎儿细胞数量已满足 PCR扩增所需的模板量 ,对胎儿性别进行预测 ,男胎符合率为 11/ 13(84.6 % ) ,性别符合率为 2 6 / 2 8(92 .9% )。结论 不连续密度梯度离心合并显微操作技术对于无创性早期产前基因诊断意义重大。
Objective To explore an effective method of obtaining fetal blood cells in maternal blood. Methods Sixty pregnant women with gestational age of 7 to 40 weeks and age of 21 to 30 years of age were subjected to discontinuous density gradient centrifugation. The isolated cells were prepared and microscopically separated by microscopy to isolate fetal nucleated erythrocytes. Y chromosome-specific DYZ1 gene by polymerase chain reaction (PCR). Results (1) The detection rate of fetal NRBC in early pregnancy was 20%. The detection rate of fetal NRBC in middle pregnancy was 71.4%. The detection rate of fetal NRBC in late pregnancy was 6.65% (P <0.05); (2) The number of fetal cells obtained by micromanipulation technique has reached the required template quantity for PCR amplification, the fetal sex is predicted, and the male fetus is consistent The rate was 11/13 (84.6%) and the gender compliance rate was 26/28 (92.9%). Conclusion Discontinuous density gradient centrifugation combined with microsurgical techniques is of great significance for the diagnosis of noninvasive early prenatal genetics.