Zhang Ling, Wang Yan, Liao Ai-Hua
Family Planning Research Institute, Center of Reproductive Medicine, Tongji Medical College, Huazhong University of Science and Technology, China.
Prenat Diagn. 2008 Dec;28(12):1160-6. doi: 10.1002/pd.2135.
To determine if quantitative abnormalities of circulating fetal trophoblast cells (CFTCs) are associated with preeclampsia.
The trophoblast cell-specific antibody, MEM-G/9 (monoclonal antibody to HLA-G), was applied to distinguish the trophoblast cells from the maternal circulation. The trophoblast cells were isolated by density-gradient centrifugation from maternal blood samples of normal pregnant and preeclamptic women, respectively. After preliminary enrichment, the CFTCs were identified by immunocytochemical staining with the MEM-G/9. To prove fetal origin of the HLA-G-positive cells, primer-extension preamplification (PEP) and polymerase chain reaction (PCR) based on single HLA-G-positive cells were adopted to detect human sex-determining region of the Y-chromosome (SRY) gene.
There were 6.88 +/- 1.54 and 30.56 +/- 5.16 HLA-G-positive cells in 6 mL maternal blood from the normal pregnant (n = 16) and preeclamptic women (n = 18), respectively. The difference was statistically significant (p < 0.001). The SRY gene from the HLA-G-positive cells was detected in all pregnant women carrying male fetuses. The sensitivity and specificity of PEP and PCR for the SRY gene detection were 100%.
It is concluded that enhancement of CFTCs numbers is related to preeclampsia, which provides information useful for noninvasive prenatal diagnosis of preeclampsia.
确定循环胎儿滋养层细胞（CFTCs）的定量异常是否与先兆子痫相关。
应用滋养层细胞特异性抗体MEM-G/9（抗HLA-G单克隆抗体）从母体循环中区分滋养层细胞。分别通过密度梯度离心从正常孕妇和先兆子痫孕妇的母血样本中分离滋养层细胞。初步富集后，用MEM-G/9进行免疫细胞化学染色鉴定CFTCs。为证明HLA-G阳性细胞的胎儿来源，采用基于单个HLA-G阳性细胞的引物延伸预扩增（PEP）和聚合酶链反应（PCR）检测Y染色体性别决定区（SRY）基因。
正常孕妇（n = 16）6 mL母血中HLA-G阳性细胞数为6.88±1.54，先兆子痫孕妇（n = 18）为30.56±5.16。差异有统计学意义（p < 0.001）。在所有怀有男性胎儿的孕妇中均检测到来自HLA-G阳性细胞的SRY基因。PEP和PCR检测SRY基因的敏感性和特异性均为100%。
得出CFTCs数量增加与先兆子痫相关的结论，这为先兆子痫的无创产前诊断提供了有用信息。
