Nondestructive Identification of Rare Trophoblastic Cells by Endoplasmic Reticulum Staining for Noninvasive Prenatal Testing of Monogenic Diseases
| Autor: | Shihua Luo, Xinyi Ye, Lei Zheng, Mei Zhong, Aifen Liang, Xiujuan Jiang, Bo Situ, Yingsi Cao, Hong Sui, Ye Zhang, Maliang Tao, Liping Huang, Yifang Huang |
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| Rok vydání: | 2020 |
| Předmět: |
trophoblastic cells
Pathology medicine.medical_specialty General Chemical Engineering Cell General Physics and Astronomy Medicine (miscellaneous) noninvasive prenatal testing 02 engineering and technology Biology 010402 general chemistry 01 natural sciences Biochemistry Genetics and Molecular Biology (miscellaneous) Genome chemistry.chemical_compound nondestructive identification medicine General Materials Science lcsh:Science Genotyping Fetus Full Paper Endoplasmic reticulum Point mutation General Engineering Full Papers monogenic diseases 021001 nanoscience & nanotechnology 0104 chemical sciences Staining medicine.anatomical_structure chemistry lcsh:Q 0210 nano-technology DNA single‐cell isolation |
| Zdroj: | Advanced Science, Vol 7, Iss 7, Pp n/a-n/a (2020) Advanced Science |
| ISSN: | 2198-3844 |
| Popis: | 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 α‐ or β‐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. An endoplasmic reticulum staining and linear discriminant analysis single‐cell test for detecting rare trophoblastic cells in Pap samples from pregnancies whose fetuses are at risk for monogenic disorders is presented. The antibody‐free, simple, low‐cost assay enables genomic analysis of fetal trophoblastic cells with high performance, which can serve as a promising tool for prenatal detection of monogenic diseases. |
| Databáze: | OpenAIRE |
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