Evidence for feasibility of fetal trophoblastic cell-based noninvasive prenatal testing.
OBJECTIVE
The goal was to develop methods for detection of chromosomal and subchromosomal abnormalities in fetal cells in the mother's circulation at 10-16 weeks' gestation using analysis by array comparative genomic hybridization (CGH) and/or next-generation sequencing (NGS).


METHODS
Nucleated cells from 30 mL of blood collected at 10-16 weeks' gestation were separated from red cells by density fractionation and then immunostained to identify cytokeratin positive and CD45 negative trophoblasts. Individual cells were picked and subjected to whole genome amplification, genotyping, and analysis by array CGH and NGS.


RESULTS
Fetal cells were recovered from most samples as documented by Y chromosome PCR, short tandem repeat analysis, array CGH, and NGS including over 30 normal male cells, one 47,XXY cell from an affected fetus, one trisomy 18 cell from an affected fetus, nine cells from a trisomy 21 case, three normal cells and one trisomy 13 cell from a case with confined placental mosaicism, and two chromosome 15 deletion cells from a case known by CVS to have a 2.7 Mb de novo deletion.


CONCLUSIONS
We believe that this is the first report of using array CGH and NGS whole genome sequencing to detect chromosomal abnormalities in fetal trophoblastic cells from maternal blood. © 2016 The Authors. Prenatal Diagnosis published by John Wiley & Sons, Ltd.
Department of Molecular and Human Genetics, Baylor College of Medicine, Houston, TX, USA.
Amy M
AM
Breman
RareCyte, Inc., Seattle, WA, USA.
Jennifer C
JC
Chow
RareCyte, Inc., Seattle, WA, USA.
Lance
L
U'Ren
Department of Molecular and Human Genetics, Baylor College of Medicine, Houston, TX, USA.
Elizabeth A
EA
Normand
Department of Molecular and Human Genetics, Baylor College of Medicine, Houston, TX, USA.
Sadeem
S
Qdaisat
Department of Molecular and Human Genetics, Baylor College of Medicine, Houston, TX, USA.
Li
L
Zhao
Department of Molecular and Human Genetics, Baylor College of Medicine, Houston, TX, USA.
David M
DM
Henke
Department of Molecular and Human Genetics, Baylor College of Medicine, Houston, TX, USA.
Rui
R
Chen
Department of Molecular and Human Genetics, Baylor College of Medicine, Houston, TX, USA.
Chad A
CA
Shaw
Department of Obstetrics and Gynecology, Drexel University College of Medicine, Philadelphia, PA, USA.
Laird
L
Jackson
Department of Molecular and Human Genetics, Baylor College of Medicine, Houston, TX, USA.
Yaping
Y
Yang
Department of Molecular and Human Genetics, Baylor College of Medicine, Houston, TX, USA.
Liesbeth
L
Vossaert
RareCyte, Inc., Seattle, WA, USA.
Rachel H V
RH
Needham
RareCyte, Inc., Seattle, WA, USA.
Elizabeth J
EJ
Chang
RareCyte, Inc., Seattle, WA, USA.
Daniel
D
Campton
RareCyte, Inc., Seattle, WA, USA.
Jeffrey L
JL
Werbin
RareCyte, Inc., Seattle, WA, USA.
Ron C
RC
Seubert
Department of Molecular and Human Genetics, Baylor College of Medicine, Houston, TX, USA.|Department of Obstetrics and Gynecology, Baylor College of Medicine, Houston, TX, USA.|Texas Children's Hospital, Houston, TX, USA.
Ignatia B
IB
Van den Veyver
RareCyte, Inc., Seattle, WA, USA.
Jackie L
JL
Stilwell
RareCyte, Inc., Seattle, WA, USA.
Eric P
EP
Kaldjian
Department of Molecular and Human Genetics, Baylor College of Medicine, Houston, TX, USA.|Texas Children's Hospital, Houston, TX, USA.
Arthur L
AL
Beaudet
D002869:Chromosome Aberrations; D055028:Comparative Genomic Hybridization; D056915:DNA Copy Number Variations; D005240:Feasibility Studies; D005260:Female; D064368:Healthy Volunteers; D006801:Humans; D008297:Male; D062145:Maternal Serum Screening Tests / Q000379:methods*; D011247:Pregnancy; D017422:Sequence Analysis, DNA; D014327:Trophoblasts / Q000166:cytology*