从孕妇外周血中捕获胎儿遗传信息的方法学探讨

Exploration to the approach of detecting fetal genetic information in maternal peripheral blood cell and plasma

目的:探讨从孕妇外周血细胞中检测胎儿基因组和血浆中捕获胎儿游离核酸方法的实用性。DNA方法:在基SRY因高保守区设计两对性别特异性引物,利用巢式聚合酶链反应分别对例孕妇外周血胎儿基因组和血浆中游(nest-PCR)50DNA离核酸进行特异性扩增,聚丙烯酰胺凝胶电泳分析,测序验证扩增产物的准确性。(PAGE)DNA结果:名孕妇中名分娩男5028婴,名分娩女婴。基因片段阳性检出率:基因组为();游离核酸组为()。名分娩女婴者除例22SRY75%21/2832%9/28221基因组检测结果阳性外,其余均为阴性结果。本研究检测系统灵敏度为52.8pg/μ。l结论:孕妇外周血中胎儿基因组和游DNA核酸检测可作为产前基因诊断的途径,两者联合应用有助于提高胎儿遗传信息的检出率。

Objective: This study was to explore the feasibility of detecting fetal genetic information in maternal peripheral blood cell and plasma. Methods: The specific fragment of SRY gene was amplified by nest-PCR in peripheral blood eukaryotic cells and plasma from 50 pregnancies. Then the PCR produces were identified by DNA sequence. Results: 28 boys and 22 girls were born from 50 pregnancies. SRY gene specific fragment was detected in maternal peripheral blood eukaryotic cell from 21 (75%) of the 28 pregnancies with male fetuses and in maternal plasma from 9 (35%) pregnancies with male fetuses. However, positive SRY signal in maternal blood eukaryotic cell was seen in only one of 22 pregnancies with female fetuses. Conclusion: Detecting fetal DNA from maternal peripheral blood could be a feasible and safe approach of non-invasive prenatal diagnosis. Our data demonstrate that it will be more effective to detect the fetal genetic information by combining analysis of eukaryotic cells and plasma from maternal peripheral blood.