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Fertil Steril. 2014 Jul;102(1):135-142.e6. doi: 10.1016/j.fertnstert.2014.04.008. Epub 2014 May 10.

Trophoblast retrieval and isolation from the cervix (TRIC) for noninvasive prenatal screening at 5 to 20 weeks of gestation.

Author information

1
Department of Obstetrics and Gynecology, C. S. Mott Center for Human Growth and Development, Wayne State University, Detroit, Michigan.
2
Department of Obstetrics and Gynecology, Georgia Regents University, Augusta, Georgia.
3
Department of Obstetrics and Gynecology, C. S. Mott Center for Human Growth and Development, Wayne State University, Detroit, Michigan; Anatomy and Cell Biology, C.S. Mott Center for Human Growth and Development, Wayne State University, Detroit, Michigan; Program in Reproductive and Adult Endocrinology, Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, Maryland. Electronic address: d.armant@wayne.edu.

Abstract

OBJECTIVE:

To use trophoblast cells accumulating in the endocervical canal at the beginning of pregnancy for noninvasive prenatal testing.

DESIGN:

Prospective, double-blinded test for fetal gender.

SETTING:

Academic medical center.

PATIENT(S):

Fifty-six women with singleton pregnancies at gestational age 5-20 weeks.

INTERVENTION(S):

Isolation of fetal cells from resident maternal cells in endocervical specimens using anti-human leukocyte antigen G coupled to magnetic nanoparticles; cell phenotyping immunofluorescently with a panel of trophoblast subtype-specific proteins; DNA integrity assessment with terminal dUTP nick-end labeling (TUNEL); and polymerase chain reaction (PCR) and fluorescent in situ hybridization (FISH) to detect sex chromosomes in individual cells.

MAIN OUTCOME MEASURE(S):

Trophoblast phenotype, TUNEL index, and percentage male cells.

RESULT(S):

The women were given a routine Papanicolaou test; fetal genders were verified from medical records. Recovery after immunomagnetic isolation averaged 746±59 cells across gestational age, with 99% expressing chorionic gonadotropin, whereas the depleted cell fraction expressed none. The isolated cells had an extravillous trophoblast phenotype and intact nuclear DNA (>95%). Fetal gender was determined in 20 specimens without error by PCR. The FISH analysis of isolated cells from male specimens validated their fetal origin.

CONCLUSION(S):

Noninvasive prenatal testing is feasible beginning at a gestational age of 5 weeks.

KEYWORDS:

Endocervical canal; fetal gender; prenatal testing; single-cell analysis; trophoblast

[Indexed for MEDLINE]

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