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Fetal-specific DNA methylation ratio permits noninvasive prenatal diagnosis of trisomy 21


The trials performed worldwide toward noninvasive prenatal diagnosis (NIPD) of Down's syndrome (or trisomy 21) have shown the commercial and medical potential of NIPD compared to the currently used invasive prenatal diagnostic procedures. Extensive investigation of methylation differences between the mother and the fetus has led to the identification of differentially methylated regions (DMRs). In this study, we present a strategy using the methylated DNA immunoprecipitation (MeDiP) methodology in combination with real-time quantitative PCR (qPCR) to achieve fetal chromosome dosage assessment, which can be performed noninvasively through the analysis of fetal-specific DMRs. We achieved noninvasive prenatal detection of trisomy 21 by determining the methylation ratio of normal and trisomy 21 cases for each tested fetal-specific DMR present in maternal peripheral blood, followed by further statistical analysis. The application of this fetal-specific methylation ratio approach provided correct diagnosis of 14 trisomy 21 and 26 normal cases.

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Figure 1: Schematic illustration of the fetal-specific DNA methylation ratio approach.
Figure 2: DNA methylation ratio values obtained from the 12 DMRs.
Figure 3: Box plot representation of the results obtained from four DMRs.


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E.A.P. and P.C.P. were supported by the Special Non-Invasive Advances in Fetal and Neonatal Evaluation Network of Excellence European Commission Funded 6th Framework Package Project Number: LSHB-CT-2004-503243 and the Cyprus Institute of Neurology and Genetics. A.K. was supported by the University of Cyprus. E.T. was supported by the State Scholarships Foundation of Greece. V.V. was supported by the Mitera Hospital in Athens. N.C. was supported by the Wellcome Trust. We thank C. Sismani, L. Kousoulidou and G. Koumbaris for extensive discussions, as well as M. Hulten for her encouragement to initiate this project.

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Authors and Affiliations



E.A.P. and E.T. carried out the experiments. E.A.P. wrote the manuscript. E.A.P. and A.K. performed the statistical analysis. E.T. and V.V. collected the majority of the samples in this study. N.P.C. provided input on the selection of the DMRs. P.C.P. was the principal investigator and has supervised the project. All authors reviewed, critiqued and offered comments on the text.

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Correspondence to Philippos C Patsalis.

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Competing interests

P.C.P. and E.A.P. have filed a US provisional patent for the fetal-specific DNA methylation ratio approach (application no. 61/405,421).

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Supplementary Tables 1–6, Supplementary Data and Supplementary Methods (PDF 174 kb)

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Papageorgiou, E., Karagrigoriou, A., Tsaliki, E. et al. Fetal-specific DNA methylation ratio permits noninvasive prenatal diagnosis of trisomy 21. Nat Med 17, 510–513 (2011).

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