Could Digital PCR Be an Alternative as a Non-Invasive Prenatal Test for Trisomy 21: A Proof of Concept Study
| Autor: | Raphaël Porcher, Dominique Le Tessier, Valérie Serazin, François Vialard, Rakia Bhouri, Laurent Mandelbrot, Audrey Coustier, Laila El Khattabi, Armelle Luscan, Christelle Rouillac Le Sciellour, Juliette Nectoux, Vassilis Tsatsaris, Jean-Michel Dupont, Thibaut Quibel |
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| Přispěvatelé: | Institut Cochin (IC UM3 (UMR 8104 / U1016)), Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Maternité Port-Royal [CHU Cochin], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Hôpital Cochin [AP-HP], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP), Département de Biologie de la Reproduction, CHI Poissy-Saint-Germain, Centre de Recherche Épidémiologie et Statistique Sorbonne Paris Cité (CRESS (U1153 / UMR_A_1125 / UMR_S_1153)), Institut National de la Recherche Agronomique (INRA)-Université Paris Diderot - Paris 7 (UPD7)-Université Paris Descartes - Paris 5 (UPD5)-Université Sorbonne Paris Cité (USPC)-Institut National de la Santé et de la Recherche Médicale (INSERM), Biochimie et biologie moléculaire, Hôpital Cochin [AP-HP], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP), Gamètes, implantation, gestation (GIG), Université de Versailles Saint-Quentin-en-Yvelines (UVSQ), Service de Gynécologie Obstétrique, Hôpital Louis Mourier, Service de Gynécologie et Obstétrique [Cochin], Assistance publique - Hôpitaux de Paris (AP-HP) (APHP)-CHU Cochin [AP-HP], Université Paris Diderot - Paris 7 (UPD7)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut National de la Recherche Agronomique (INRA)-Université Paris Descartes - Paris 5 (UPD5)-Université Sorbonne Paris Cité (USPC), CHU Cochin [AP-HP], Institut Cochin (UM3 (UMR 8104 / U1016)), Université Paris Descartes - Paris 5 (UPD5) - Institut National de la Santé et de la Recherche Médicale (INSERM) - Centre National de la Recherche Scientifique (CNRS), Assistance publique - Hôpitaux de Paris (AP-HP) - CHU Cochin [AP-HP], Centre de Recherche Épidémiologie et Statistique Sorbonne Paris Cité (CRESS (U1153 / UMR_A 1125)), Institut National de la Recherche Agronomique (INRA) - Université Sorbonne Paris Cité (USPC) - Institut National de la Santé et de la Recherche Médicale (INSERM), Unité de Pathologie Cellulaire et Génétique (UPRES-EA2493), Bos, Mireille, Département de biologie de la reproduction et de gynécologie [CHIPS, Poissy], Centre hospitalier intercommunal de Poissy/Saint-Germain-en-Laye - CHIPS [Poissy] |
| Jazyk: | angličtina |
| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
Chromosomes Human Pair 21 Maternal Health lcsh:Medicine Artificial Gene Amplification and Extension Polymerase Chain Reaction law.invention Chromosomal Disorders 0302 clinical medicine Pregnancy law Prenatal Diagnosis Medicine and Health Sciences Digital polymerase chain reaction lcsh:Science Polymerase chain reaction ComputingMilieux_MISCELLANEOUS Genetics 030219 obstetrics & reproductive medicine Multidisciplinary Chromosome Biology Obstetrics and Gynecology 3. Good health Chromosome 21 Karyotypes DNA Probes Research Article Down syndrome Validation study Computational biology Biology Research and Analysis Methods Chromosomes Cytogenetics 03 medical and health sciences Multiplex polymerase chain reaction [SDV.BBM] Life Sciences [q-bio]/Biochemistry Molecular Biology medicine Humans [SDV.BBM]Life Sciences [q-bio]/Biochemistry Molecular Biology Molecular Biology Techniques Molecular Biology Clinical Genetics Trisomics lcsh:R Non invasive Biology and Life Sciences Reproducibility of Results Cell Biology DNA Aneuploidy Chromosome Pairs medicine.disease 030104 developmental biology ROC Curve Women's Health lcsh:Q Down Syndrome Trisomy Departures from Diploidy Multiplex Polymerase Chain Reaction |
| Zdroj: | PLoS ONE PLoS ONE, Public Library of Science, 2016, ⟨10.1371/journal.pone.0155009⟩ PLoS ONE, Vol 11, Iss 5, p e0155009 (2016) PLoS ONE, Public Library of Science, 2016, 〈10.1371/journal.pone.0155009〉 PLoS ONE, 2016, ⟨10.1371/journal.pone.0155009⟩ |
| ISSN: | 1932-6203 |
| Popis: | Objective NIPT for fetal aneuploidy by digital PCR has been hampered by the large number of PCR reactions needed to meet statistical requirements, preventing clinical application. Here, we designed an octoplex droplet digital PCR (ddPCR) assay which allows increasing the number of available targets and thus overcomes statistical obstacles. Method After technical optimization of the multiplex PCR on mixtures of trisomic and euploid DNA, we performed a validation study on samples of plasma DNA from 213 pregnant women. Molecular counting of circulating cell-free DNA was performed using a mix of hydrolysis probes targeting chromosome 21 and a reference chromosome. Results The results of our validation experiments showed that ddPCR detected trisomy 21 even when the sample’s trisomic DNA content is as low as 5%. In a validation study of plasma samples from 213 pregnant women, ddPCR discriminated clearly between the trisomy 21 and the euploidy groups. Conclusion Our results demonstrate that digital PCR can meet the requirements for non-invasive prenatal testing of trisomy 21. This approach is technically simple, relatively cheap, easy to implement in a diagnostic setting and compatible with ethical concerns regarding access to nucleotide sequence information. These advantages make it a potential technique of choice for population-wide screening for trisomy 21 in pregnant women. |
| Databáze: | OpenAIRE |
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