Objective: To establish a method for noninvasive fetal cell isolation from maternal blood and prenatal testing of monogenic diseases by a combination of direct sequencing and targeted NGS-based SNP haplotyping from single fetal cells. Method: Peripheral blood of pregnant women in two families (congenital deafness and ichthyosis) was collected. After density-based separation and immunostaining with multiple biomarkers, candidate fetal cells were identified by high-throughput imagine analysis and picked up by automation. Individual fetal cells were subjected to STR-genotyping to identify their origin. Pathogenic mutations were identified by direct Sanger sequencing, and a combination of targeted NGS and SNP haplotyping using a custom panel. All the results were compared with amniotic fluid DNA. Results: Fetal trophoblasts were successfully harvested from maternal blood. STRgenotyping confirmed the fetal origin. Direct sequencing of pathogenic genetic mutations in fetal cells showed consistent results with amniotic fluid samples. For congenital deafness family, NGS-based SNP haplotyping also correctly identified the fetal haplotype. This single cell haplotyping method can be used to diagnose various genetic diseases. Conclusion: We have established a method for noninvasive prenatal testing of monogenic diseases from circulating trophoblast cells. This cell-based NIPT can be further applied to the prenatal diagnosis of various monogenic diseases.

Abstract.We study the problem of flatness of two-input driftless control systems. Although a characterization of flat systems of that class is known, the problems of describing all flat outputs and of calculating them is open and we solve it in the paper. We show that all x-flat outputs are parameterized by an arbitrary function of three canonically defined variables. We also construct a system of 1st order PDE's whose solutions give all x-flat outputs of two-input driftless systems. We illustrate our results by describing all x-flat outputs of models of a nonholonomic car and the n-trailer system.Mathematics Subject Classification. 93B27, 93C10, 93C95.

No abstract