A novel method for noninvasive diagnosis of monogenic diseases from circulating fetal cells.

Prenatal Diagnosis
Liang ChangPing Liu

Abstract

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. 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. Fetal trophoblasts were successfully harvested from maternal blood. STR-genotyping 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. We have established a method for noninva...Continue Reading

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Citations

Mar 7, 2021·International Journal of Molecular Sciences·Silvia SpenaFlora Peyvandi
Mar 8, 2021·Journal of Assisted Reproduction and Genetics·Christian Liebst Frisk ToftInge Søkilde Pedersen
May 12, 2021·Prenatal Diagnosis·Liesbeth VossaertIgnatia B Van den Veyver

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