Wayne State University

D Randall Armant

Sascha Drewlo

Michael P Diamond

Methods of isolating DNA of a fetus of an ongoing pregnancy are provided according to the present invention which include obtaining a maternal endocervical sample containing maternal cells and fetal extravillous trophoblast cells from a pregnant subject; isolating fetal extravillous trophoblast cells from the maternal endocervical sample, producing isolated fetal extravillous trophoblast cells contaminated with maternal DNA; lysing the isolated fetal extravillous trophoblast cells; isolating fetal nuclei from the lysed fetal extravillous trophoblast cells, producing isolated fetal nuclei, thereby removing at least a portion of the contaminating maternal DNA; and purifying genomic DNA from the isolated fetal nuclei, producing purified fetal genomic DNA. The purified fetal genomic DNA can be assayed to determine a characteristic of a genomic DNA sequence of the purified fetal genomic DNA, thereby determining a characteristic of DNA of a fetus of an ongoing pregnancy.

Isolation and analysis of fetal DNA from extravillous trophoblast cells retrieved from the endocervical canal

A method of retrieving fetal cells from an endocervical sample by removing the mucus from the endocervical sample by disassociating fetal cells and maternal cells in the endocervical sample; and isolating disassociated fetal cells from other cells in the endocervical sample. Also provided is a method of retrieving fetal cells from an endocervical sample, by obtaining a mixture of disassociated cells prepared by the above method, treating the cells with a fetal-specific antibody, identifying cells that have bound to the fetal-specific antibody, and isolating the identified cells. The disassociated cell prepared by the above method can be analyzed and used for a variety of purposes including, but not limited to, the identification of fetal cells among cervical cells, determination of fetal cell density to predict high risk pregnancy, genetic analysis of fetal cells, and determination of growth factor or other biomarker expression to predict obstetrical disorders.

Identification and analysis of fetal trophoblast cells in cervical mucus for prenatal diagnosis

Methods of isolating and assaying fetal extravillous trophoblast cells, including assays of RNA of the fetal extravillous trophoblast cells according to aspects of the disclosure include obtaining a maternal endocervical sample containing fetal extravillous trophoblast cells from a pregnant subject; fixing the maternal endocervical sample in an aldehyde fixative, removing fetal extravillous trophoblast cells from the maternal endocervical sample thereby producing isolated extravillous trophoblast cells, isolating and assaying RNA from the fetal extravillous trophoblast cells.

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Saint Clair Shores, MI, United States of America

D Randall Armant