用于确定新型非侵入性产前诊断方法胎儿细胞富集效率的体内模型。

In vivo model to determine fetal-cell enrichment efficiency of novel noninvasive prenatal diagnosis methods.

作者信息

Ponnusamy S, Mohammed N, Ho S S Y, Zhang H M, Chan Y H, Ng Y W, Su L L, Mahyuddin A P, Venkat A, Chan J, Rauff M, Biswas A, Choolani M

机构信息

Diagnostic Biomarker Discovery Laboratory, Division of Maternal and Fetal Medicine, Department of Obstetrics and Gynecology, Yong Loo Lin School of Medicine, National University of Singapore, Singapore.

出版信息

Prenat Diagn. 2008 Jun;28(6):494-502. doi: 10.1002/pd.2009.

DOI:10.1002/pd.2009

PMID:18509867

Abstract

OBJECTIVE

To develop an in vivo model to determine fetal-cell enrichment efficiency of novel noninvasive prenatal diagnosis methods.

METHODS

Efficiency of our three-step enrichment protocol was determined in vitro before fetal nucleated red blood cells (FNRBCs) were enriched from first-trimester maternal blood samples collected from the same patients pre- and postsurgical termination of pregnancy (TOP) (n = 10). FNRBCs enriched were identified using embryonic epsilon-globin immunocytochemistry and chromosomal fluorescence in situ hybridization.

RESULTS

We recovered 37% of spiked FNRBCs (95% confidence interval (CI) 28.5-45.6; n = 8) in in vitro experiments. We show a consistent threefold increase in the number of epsilon + FNRBCs in maternal blood obtained immediately post-TOP (p = 0.005). A mathematical relationship was derived: observed number of pretermination primitive FNRBCs = 0.6 + 0.31 (coefficient between pretermination/post-termination primitive FNRBCs, 95% CI 0.12-0.49; p = 0.005) x observed number of post-termination primitive FNRBCs (R2 = 0.65).

CONCLUSION

Our data demonstrate that maternal blood obtained immediately post-TOP would be a good in vivo model to determine the enrichment efficiency of novel protocols and methods for noninvasive prenatal diagnosis.

摘要

目的

建立一种体内模型，以确定新型非侵入性产前诊断方法的胎儿细胞富集效率。

方法

在从同一患者妊娠终止手术（TOP）前后收集的孕早期母血样本中富集胎儿有核红细胞（FNRBCs）之前，先在体外确定我们三步富集方案的效率（n = 10）。使用胚胎ε-珠蛋白免疫细胞化学和染色体荧光原位杂交鉴定富集的FNRBCs。

结果

在体外实验中，我们回收了37%的加标的FNRBCs（95%置信区间（CI）28.5 - 45.6；n = 8）。我们发现TOP后立即获得的母血中ε + FNRBCs数量一致增加了三倍（p = 0.005）。得出了一种数学关系：终止前原始FNRBCs的观察数量 = 0.6 + 0.31（终止前/终止后原始FNRBCs之间的系数，95% CI 0.12 - 0.49；p = 0.005）×终止后原始FNRBCs的观察数量（R2 = 0.65）。