Nondestructive Identification of Rare Trophoblastic Cells by Endoplasmic Reticulum Staining for Noninvasive Prenatal Testing of Monogenic Diseases
Nondestructive Identification of Rare Trophoblastic Cells by Endoplasmic Reticulum Staining for Noninvasive Prenatal Testing of Monogenic Diseases
复制标题
内质网染色无损鉴定稀有滋养层细胞，用于单基因疾病的无创产前检测
DOI：
10.1002/advs.201903354
复制
发表时间：
2020-02-13
期刊：
影响因子：
15.1
通讯作者：
Zheng, Lei
中科院分区：
文献类型：
--
作者：
Huang, Yifang;Situ, Bo;Zheng, Lei
基于无细胞DNA的单基因疾病的无创产前检测由于难以获得足够比例和足够质量的胎儿DNA而受到阻碍。分析携带整个胎儿基因组的Papanicolaou涂片中罕见的滋养细胞为单基因疾病的无创检测提供了另一种方法。然而，细胞内标记对靶细胞的鉴定会不同程度地影响DNA的质量。本文建立了一种基于内质网染色和线性判别分析（ER-LDA）的从丰富的母细胞中无损鉴定稀有胎儿细胞的新方法。与传统方法相比，ER-LDA对细胞质量影响很小，可以在单细胞水平上分析滋养细胞。使用ER-LDA，可以在单细胞分辨率下从60例妊娠4至38周的孕妇中鉴定和分离高纯度的滋养细胞。在11个有α -或β -地中海贫血风险的胎儿中，包括-(SEA)/缺失突变和点突变在内的致病变异可以通过该测试准确检测出来。该检测平台还可以扩展到分析其他单基因疾病的突变谱。这种简单、低成本、无创的检测方法可以为胎儿基因分型提供有价值的胎儿细胞，并有望用于单基因疾病的产前检测。
Noninvasive prenatal detection of monogenic diseases based on cell-free DNA is hampered by challenges in obtaining a sufficient fraction and adequate quality of fetal DNA. Analyzing rare trophoblastic cells from Papanicolaou smears carrying the entire fetal genome provides an alternative method for noninvasive detection of monogenic diseases. However, intracellular labeling for identification of target cells can affect the quality of DNA in varying degrees. Here, a new approach is developed for nondestructive identification of rare fetal cells from abundant maternal cells based on endoplasmic reticulum staining and linear discriminant analysis (ER-LDA). Compared with traditional methods, ER-LDA has little effect on cell quality, allowing trophoblastic cells to be analyzed on the single-cell level. Using ER-LDA, high-purity of trophoblastic cells can be identified and isolated at single cell resolution from 60 pregnancies between 4 and 38 weeks of gestation. Pathogenic variants, including -(SEA)/ deletion mutation and point mutations, in 11 fetuses at risk for alpha- or beta-thalassemia can be accurately detected by this test. The detection platform can also be extended to analyze the mutational profiles of other monogenic diseases. This simple, low-cost, and noninvasive test can provide valuable fetal cells for fetal genotyping and holds promise for prenatal detection of monogenic diseases.