Objective: To investigate potential performance of cell-free DNA (cfDNA) testing in maternal blood in detecting fetal triploidy. Methods: Plasma and buffy coat samples obtained at 11-13 weeks' gestation from singleton pregnancies with diandric triploidy (n = 4), digynic triploidy (n = 4), euploid fetuses (n = 48) were sent to Natera, Inc. (San Carlos, Calif., USA) for cfDNA testing. Multiplex polymerase chain reaction amplification of cfDNA followed by sequencing of single nucleotide polymorphic loci covering chromosomes 13, 18, 21, X, and Y was performed. Sequencing data were analyzed using the NATUS algorithm which identifies copy number for each of the five chromosomes. Results: cfDNA testing provided a result in 44 (91.7%) of the 48 euploid cases and correctly predicted the fetal sex and the presence of two copies each of chromosome 21, 18 and 13. In diandric triploidy, cfDNA testing identified multiple paternal haplotypes (indicating fetal trisomy 21, trisomy 18 and trisomy 13) suggesting the presence of either triploidy or dizygotic twins. In digynic triploidy the fetal fraction corrected for maternal weight and gestational age was below the 0.5th percentile. Conclusions: cfDNA testing by targeted sequencing and allelic ratio analysis of single nucleotide polymorphisms covering chromosomes 21, 18, 13, X, and Y can detect diandric triploidy and raise the suspicion of digynic triploidy.

Keywords:
Non-invasive diagnosis, Prenatal diagnosis, Triploidy, Cell-free DNA
1.
Jacobs PA, Angell RR, Buchanan IM, Hassold TJ, Matsuyama JS, Manuel B: The origin of human triploids. Ann Hum Genet 1978;42:49-57.
2.
Snijders RJM, Sebire NJ, Nicolaides KH: Maternal age and gestational age-specific risk for chromosomal defects. Fetal Diagn Ther 1995;10:356-367.
3.
McFadden DE, Kwong LC, Yam IYL, Langlois S: Parental origin of triploidy in human fetuses: evidence for genomic imprinting. Hum Genet 1993;92:465-469.
4.
Jauniaux E, Brown R, Snijders RJ, Noble P, Nicolaides KH: Early prenatal diagnosis of triploidy. Am J Obstet Gynecol 1997;176:550-554.
5.
Spencer K, Liao AW, Skentou H, Cicero S, Nicolaides KH: Screening for triploidy by fetal nuchal translucency and maternal serum free β-hCG and PAPP-A at 10-14 weeks of gestation. Prenat Diagn 2000;20:495-499.
6.
Kagan KO, Anderson JM, Anwandter G, Neksasova K, Nicolaides KH: Screening for triploidy by the risk algorithms for trisomies 21, 18 and 13 at 11 weeks to 13 weeks and 6 days of gestation. Prenat Diagn 2008;28:1209-1213.
7.
Rijhsinghani A, Yankowitz J, Strauss RA, Kuller JA, Patil S, Williamson RA: Risk of preeclampsia in second-trimester triploid pregnancies. Obstet Gynecol 1997;90:884-888.
8.
Seckl MJ, Sebire NJ, Berkowitz RS: Gestational trophoblastic disease. Lancet 2010;376:717-729.
9.
Engelbrechtsen L, Brøndum-Nielsen K, Ekelund C, Tabor A, Skibsted L: Detection of triploidy at 11-14 weeks of gestation: a cohort study of 198,000 pregnant women. Ultrasound Obstet Gynecol 2013, E-pub ahead of print.
10.
Palomaki GE, Kloza EM, Lambert-Messerlian GM, Haddow JE, Neveux LM, Ehrich M, van den Boom D, Bombard AT, Deciu C, Grody WW, Nelson SF, Canick JA: DNA sequencing of maternal plasma to detect Down syndrome: an international clinical validation study. Genet Med 2011;13:913-920.
11.
Bianchi DW, Platt LD, Goldberg JD, Abuhamad AZ, Sehnert AJ, Rava RP: Genome-wide fetal aneuploidy detection by maternal plasma DNA sequencing. Obstet Gynecol 2012;119:890-901.
12.
Sparks AB, Struble CA, Wang ET, Song K, Oliphant A: Noninvasive prenatal detection and selective analysis of cell-free DNA obtained from maternal blood: evaluation for trisomy 21 and trisomy 18. Am J Obstet Gynecol 2012;206:319.e1-e9.
13.
Ashoor G, Syngelaki A, Wagner M, Birdir C, Nicolaides KH: Chromosome-selective sequencing of maternal plasma cell-free DNA for first-trimester detection of trisomy 21 and trisomy 18. Am J Obstet Gynecol 2012;206:322.e1-e5.
14.
Nicolaides KH, Syngelaki A, Ashoor G, Birdir C, Touzet G: Noninvasive prenatal testing for fetal trisomies in a routinely screened first-trimester population. Am J Obstet Gynecol 2012;207:374.e1-e6.
15.
Zimmermann B, Hill M, Gemelos G, Demko Z, Banjevic M, Baner J, Ryan A, Sigurjonsson S, Chopra N, Dodd M, Levy B, Rabinowitz M: Noninvasive prenatal aneuploidy testing of chromosomes 13, 18, 21, X, and Y using targeted sequencing of polymorphic loci. Prenat Diag 2012;32:1233-1241.
16.
Nicolaides KH, Syngelaki A, Gil M, Atanasova V, Markova D: Validation of targeted sequencing of single-nucleotide polymorphisms for non-invasive prenatal detection of aneuploidy of chromosomes 13, 18, 21, X, and Y. Prenat Diagn 2013;33:575-579.
17.
Samango-Sprouse C, Banjevic M, Ryan A, Sigurjonsson S, Zimmermann B, Hill M, Hall MP, Westemeyer M, Saucier J, Demko Z, Rabinowitz M: SNP-based non-invasive prenatal testing detects sex chromosome aneuploidies with high accuracy. Prenat Diagn 2013;33:643-649.
18.
Kagan KO, Wright D, Valencia C, Maiz N, Nicolaides KH: Screening for trisomies 21, 18 and 13 by maternal age, fetal nuchal translucency, fetal heart rate, free β-hCG and pregnancy-associated plasma protein-A. Hum Reprod 2008;23:1968-1975.
19.
Liao GJ, Chan KC, Jiang P, Sun H, Leung TY, Chiu RWK, Lo DYM: Non-invasive prenatal diagnosis of fetal trisomy 21 by allelic ratio analysis using targets massively parallel sequencing of maternal plasma DNA. PLoS One 2012;7:e38154.
20.
Ashoor G, Poon L, Syngelaki A, Mosimann B, Nicolaides KH: Fetal fraction in maternal plasma cell-free DNA at 11-13 weeks' gestation: effect of maternal and fetal factors. Fetal Diagn Ther 2012;31:237-243.
21.
Ashoor G, Syngelaki A, Poon LC, Rezende JC, Nicolaides KH: Fetal fraction in maternal plasma cell-free DNA at 11-13 weeks' gestation: relation to maternal and fetal characteristics. Ultrasound Obstet Gynecol 2013;41:26-32.
22.
Poon LC, Misci T, Song K, Syngelaki A, Nicolaides KH: Maternal plasma cell-free DNA at 11-13 weeks' gestation: relation to fetal and maternal characteristics and pregnancy outcomes. Fetal Diagn Ther 2013;33:215-223.
23.
Haghiac M, Vora NL, Basu S, Johnson KL, Presley L, Bianchi DW, Mouzon SH: Increased death of adipose cells, a path to release cell-free DNA into systemic circulation of obese women. Obesity (Silver Spring) 2012;20:2213-2219.
24.
Faas BH, de Ligt J, Janssen I, Eggink AJ, Wijnberger LD, van Vugt JM, Vissers L, Geurts van Kessel A: Non-invasive prenatal diagnosis of fetal aneuploidies using massively parallel sequencing-by-ligation and evidence that cell-free fetal DNA in the maternal plasma originates from cytotrophoblastic cells. Expert Opin Biol Ther 2012;12(suppl 1):S19-S26.
25.
Gil MM, Quezada MS, Bregant B, Ferraro M, Nicolaides KH: Implementation of maternal blood cell-free DNA testing in early screening for aneuploidies. Ultrasound Obstet Gynecol 2013;42:34-40.
26.
Nicolaides KH, Wright D, Poon LC, Syngelaki A, Gil MM: First-trimester contingent screening for trisomy 21 by biomarkers and maternal blood cell-free DNA testing. Ultrasound Obstet Gynecol 2013;42:41-50.
27.
Nicolaides KH: Turning the pyramid of prenatal care. Fetal Diagn Ther 2011;29:183- 196.
© 2013 S. Karger AG, Basel
2013
Copyright / Drug Dosage / Disclaimer
Copyright: All rights reserved. No part of this publication may be translated into other languages, reproduced or utilized in any form or by any means, electronic or mechanical, including photocopying, recording, microcopying, or by any information storage and retrieval system, without permission in writing from the publisher.
Drug Dosage: The authors and the publisher have exerted every effort to ensure that drug selection and dosage set forth in this text are in accord with current recommendations and practice at the time of publication. However, in view of ongoing research, changes in government regulations, and the constant flow of information relating to drug therapy and drug reactions, the reader is urged to check the package insert for each drug for any changes in indications and dosage and for added warnings and precautions. This is particularly important when the recommended agent is a new and/or infrequently employed drug.
Disclaimer: The statements, opinions and data contained in this publication are solely those of the individual authors and contributors and not of the publishers and the editor(s). The appearance of advertisements or/and product references in the publication is not a warranty, endorsement, or approval of the products or services advertised or of their effectiveness, quality or safety. The publisher and the editor(s) disclaim responsibility for any injury to persons or property resulting from any ideas, methods, instructions or products referred to in the content or advertisements.
You do not currently have access to this content.