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目的评价使用孕妇外周血中的胎儿细胞和游离DNA进行无创伤性产前诊断的可能性。方法研究对象为150名孕5～9周的孕妇。使用淋巴细胞分离液和3%的明胶分离胎儿细胞。使用荧光定量PCR(FISH)检测母血中胎儿细胞Y染色体的末端。同时使用巢氏PCR和荧光定量PCR(FQ- PCR)检测150名孕妇血浆中的胎儿SRY基因。结果FISH、巢氏PCR以及FQ-PCR 3种方法的敏感性分别为73．2%、78%和85．4%,特异性分别为96．2%、100%和100%。使用FISH,最早在孕49d检测到胎儿细胞,使用巢氏PCR和FQ-PCR最早在孕49d和42d检测到孕妇血浆中的胎儿游离DNA。结论3种方法对无创伤性产前诊断都是适合的,其中FQ-PCR因其检测时间较早而成为最优选择。
Objective To evaluate the possibility of noninvasive prenatal diagnosis using fetal cells and free DNA in pregnant women’s peripheral blood. Methods Study of 150 pregnant women 5-9 weeks pregnant women. Fetal cells were isolated using lymphocyte fraction and 3% gelatin. Fluorescent quantitative PCR (FISH) was used to detect the end of the fetal cell Y chromosome in maternal blood. The fetal SRY gene in plasma of 150 pregnant women was detected by nested PCR and real-time quantitative PCR (FQ-PCR). Results The sensitivity of FISH, nest PCR and FQ-PCR were 73.2%, 78% and 85.4% respectively, and the specificity were 96.2%, 100% and 100% respectively. FISH was used to detect fetal cells as early as 49 days of gestation. Fetal DNA was detected in maternal plasma as early as 49 days and 42 days by using nested PCR and FQ-PCR. Conclusion All three methods are suitable for noninvasive prenatal diagnosis. Among them, FQ-PCR is the best choice because of its earlier detection time.