Abstract
Congenital adrenal hyperplasia (CAH) is an autosomal recessive defect in cortisol biosynthesis that elevates fetal androgen levels to cause genital ambiguity and external genital masculinization in newborn females. Introducing dexamethasone in utero by 7 weeks gestation precludes virilization of affected females. However, identification of a male fetus prior to week 7 could avert the necessity of steroid treatment in half of pregnancies at risk of CAH. We recently introduced trophoblast retrieval and isolation from the cervix (TRIC), an approach that noninvasively isolate homogeneous trophoblast cells from pregnant women as early as 5 weeks gestation, using a Papanicolaou test. Here, we have used TRIC to correctly identify male fetal DNA when both parents were carriers of the mutation that produces CAH and previously produced an affected child. Trophoblast cells (1400) obtained by TRIC were assessed using immunocytochemistry with an antibody against the trophoblast-specific β subunit of human chorionic gonadotropin, which labeled 100% (17 of 17) of isolated cells, while none of the excluded maternal cervical cells were labeled. The isolated cells were examined by fluorescent in situ hybridization for chromosomes 18, X, and Y at a clinical cytogenetics laboratory, demonstrating 100% (18 of 18) of cells to be diploid 18/XY. Aliquots of DNA obtained from the isolated cells assayed for SRY and RNASEH genes by TaqMan assays confirmed a male fetus. This case study demonstrates the utility of TRIC to accurately identify fetal gender as a means of reducing the need for prophylactic administration of exogenous steroids in pregnancies at risk of CAH.
Similar content being viewed by others
References
Nimkarn S, New MI. Congenital adrenal hyperplasia due to 21-hydroxylase deficiency: a paradigm for prenatal diagnosis and treatment. Ann N Y Acad Sci. 2010;1192:5–11.
Auchus RJ. The Classic and Nonclassic Congenital Adrenal Hyperplasias. Endocr Pract. 2015;21(4): 383–389.
Merke DP, Bornstein SR. Congenital adrenal hyperplasia. Lancet. 2005;365(9477):2125–2136.
Witchel SF, Miller WL. Prenatal treatment of congenital adrenal hyperplasia-not standard of care. J Genet Couns. 2012;21(5): 615–624.
Raff H, Sharma ST, Nieman LK. Physiological basis for the etiology, diagnosis, and treatment of adrenal disorders: Cushing’s syndrome, adrenal insufficiency, and congenital adrenal hyperplasia. Compr Physiol. 2014;4(2):739–769.
Lajic S, Nordenstrom A, Ritzen EM, Wedell A. Prenatal treatment of congenital adrenal hyperplasia. Eur J Endocrinol. 2004; 151(suppl 3):u63–u69.
Gidlof S, Wedell A, Guthenberg C, von Dobeln U, Nordenstrom A. Nationwide neonatal screening for congenital adrenal hyperplasia in sweden: a 26-year longitudinal prospective population-based study. JAMA Pediatr. 2014;168(6):567–574.
Dreger A, Feder EK, Tamar-Mattis A. Prenatal Dexamethasone for Congenital Adrenal Hyperplasia: An Ethics Canary in the Modern Medical Mine. J Bioeth Inq. 2012;9(3):277–294.
Merce Fernandez-Balsells M, Muthusamy K, Smushkin G, et al. Prenatal dexamethasone use for the prevention of virilization in pregnancies at risk for classical congenital adrenal hyperplasia because of 21-hydroxylase (CYP21A2) deficiency: a systematic review and meta-analyses. Clin Endocrinol (Oxf). 2010;73(4): 436–444.
Speiser PW, Azziz R, Baskin LS, et al. Congenital adrenal hyperplasia due to steroid 21-hydroxylase deficiency: an Endocrine Society clinical practice guideline. J Clin Endocrinol Metab. 2010;95(9):4133–4160.
Forest MG. Recent advances in the diagnosis and management of congenital adrenal hyperplasia due to 21-hydroxylase deficiency. Hum Reprod Update. 2004;10(6):469–485.
Miller WL, Witchel SF. Prenatal treatment of congenital adrenal hyperplasia: risks outweigh benefits. Am J Obstet Gynecol. 2013; 208(5):354–359.
Altarescu G, Barenholz O, Renbaum P, et al. Preimplantation genetic diagnosis (PGD)–prevention of the birth of children affected with endocrine diseases. J Pediatr Endocrinol Metab. 2011;24(7–8):543–548.
Lo YM. Non-invasive prenatal diagnosis by massively parallel sequencing of maternal plasma DNA. Open Biol. 2012;2(6): 120086.
New MI, Tong YK, Yuen T, et al. Noninvasive prenatal diagnosis of congenital adrenal hyperplasia using cell-free fetal DNA in maternal plasma. J Clin Endocrinol Metab. 2014;99(6): e1022–e1030.
Bianchi DW, Parker RL, Wentworth J, et al. DNA sequencing versus standard prenatal aneuploidy screening. N Engl J Med. 2014;370(9):799–808.
Gil MM, Quezada MS, Revello R, Akolekar R, Nicolaides KH. Analysis of cell-free DNA in maternal blood in screening for fetal aneuploidies: updated meta-analysis. Ultrasound Obstet Gynecol. 2015;45(3):249–266.
Lo YM. Non-invasive prenatal diagnosis by massively parallel sequencing of maternal plasma DNA. Open Biol. 2012;2(6): 120086.
Wang E, Batey A, Struble C, Musci T, Song K, Oliphant A. Gestational age and maternal weight effects on fetal cell-free DNA in maternal plasma. Prenat Diagn. 2013;33(7):662–666.
Van der Aa N, Zamani Esteki M, Vermeesch JR, Voet T. Preimplantation genetic diagnosis guided by single-cell genomics. Genome Med. 2013;5(8):71.
Dahdouh EM, Balayla J, Audibert F, et al. Technical update: preimplantation genetic diagnosis and screening. J Obstet Gynaecol Can. 2015;37(5):451–463.
Berger VK, Baker VL. Preimplantation diagnosis for single gene disorders. Semin Reprod Med. 2014;32(2):107–113.
Imudia AN, Kumar S, Diamond MP, Decherney AH, Armant DR. Transcervical retrieval of fetal cells in the practice of modern medicine: a review of the current literature and future direction. Fertil Steril. 2010;93(6):1725–1730.
Imudia AN, Suzuki Y, Kilburn BA, et al. Retrieval of trophoblast cells from the cervical canal for prediction of abnormal pregnancy: a pilot study. Hum Reprod. 2009;24(9):2086–2092.
Bolnick JM, Kilburn BA, Bajpayee S, et al. Trophoblast retrieval and isolation from the cervix (TRIC) for noninvasive prenatal screening at 5 to 20 weeks of gestation. Fertil Steril. 2014; 102(1):135–142. e136.
Fritz R, Kohan-Ghadr HR, Sacher A, et al. Trophoblast Retrieval and Isolation from the Cervix (TRIC) is unaffected by early gestational age or maternal obesity. Prenat Diagn. 2015;35(12): 1218–1222.
Orr JW Jr, Barrett JM, Orr PF, Holloway RW, Holimon JL. The efficacy and safety of the cytobrush during pregnancy. Gynecol Oncol. 1992;44(3):260–262.
Rivlin ME, Woodliff JM, Bowlin RB, et al. Comparison of cytobrush and cotton swab for Papanicolaou smears in pregnancy. J Reprod Med. 1993;38(2):147–150.
Paraiso MF, Brady K, Helmchen R, Roat TW. Evaluation of the endocervical Cytobrush and Cervex-Brush in pregnant women. Obstet Gynecol. 1994;84(4):539–543.
Foster JC, Smith HL. Use of the Cytobrush for Papanicolaou smear screens in pregnant women. J Nurse Midwifery. 1996; 41(3):211–217.
Holt J, Stiltner L, Jamieson B, Fashner J. Clinical inquiries. Should a nylon brush be used for Pap smears from pregnant women? J Fam Pract. 2005;54(5):463–464.
Loke YW, King A, Burrows T, et al. Evaluation of trophoblast HLA-G antigen with a specific monoclonal antibody. Tissue Antigens. 1997;50(2):135–146.
American College of Obstetricians and Gynecologists. ACOG Practice Bulletin No. 88, December 2007. Invasive prenatal testing for aneuploidy. Obstet Gynecol. 2007;110(6):1459–1467.
Pang S, Clark AT, Freeman LC, et al. Maternal side effects of prenatal dexamethasone therapy for fetal congenital adrenal hyperplasia. J Clin Endocrinol Metab. 1992;75(1):249–253.
Ritzen EM. Prenatal dexamethasone treatment of fetuses at risk for congenital adrenal hyperplasia: benefits and concerns. Semin Neonatol. 2001;6(4):357–362.
Lekarev O, New MI. Adrenal disease in pregnancy. Best Pract Res Clin Endocrinol Metab. 2011;25(6):959–973.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Bolnick, A.D., Fritz, R., Jain, C. et al. Trophoblast Retrieval and Isolation From the Cervix for Noninvasive, First Trimester, Fetal Gender Determination in a Carrier of Congenital Adrenal Hyperplasia. Reprod. Sci. 23, 717–722 (2016). https://doi.org/10.1177/1933719116632922
Published:
Issue Date:
DOI: https://doi.org/10.1177/1933719116632922