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Plasma placental RNA allelic ratio permits noninvasive prenatal chromosomal aneuploidy detection

Nature Medicine volume 13, pages 218–223 (2007)Cite this article

Abstract

Current methods for prenatal diagnosis of chromosomal aneuploidies involve the invasive sampling of fetal materials using procedures such as amniocentesis or chorionic villus sampling and constitute a finite risk to the fetus. Here, we outline a strategy for fetal chromosome dosage assessment that can be performed noninvasively through analysis of placental expressed mRNA in maternal plasma. We achieved noninvasive prenatal diagnosis of fetal trisomy 21 by determining the ratio between alleles of a single-nucleotide polymorphism (SNP) in PLAC4 mRNA, which is transcribed from chromosome 21 and expressed by the placenta, in maternal plasma. PLAC4 mRNA in maternal plasma was fetal derived and cleared after delivery. The allelic ratios in maternal plasma correlated with those in the placenta. Fetal trisomy 21 was detected noninvasively in 90% of cases and excluded in 96.5% of controls.

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Figure 1: Schematic of the RNA-SNP allelic ratio strategy for noninvasive detection of fetal trisomy 21 through maternal plasma analysis by the relative quantification of alleles of placenta-expressed transcripts located on chromosome 21.
Figure 2: Detection of PLAC4 mRNA in maternal plasma.
Figure 3: Schematic illustration of the analytical approach for RNA-SNP allelic ratio determination.
Figure 4: PLAC4 mRNA for fetal trisomy-21 detection.
Figure 5: The relative insensitivity of RNA-SNP ratio analysis to gestational age and placental mosaicism.

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Acknowledgements

This project was supported by the Innovation and Technology Fund of the Hong Kong SAR Government (ITS/195/01) and the Li Ka Shing Foundation.

Author information

Author notes

  1. Y M Dennis Lo, Nancy B Y Tsui, Rossa W K Chiu and Chunming Ding: These authors contributed equally to this work.

Authors and Affiliations

  1. Li Ka Shing Institute of Health Sciences, The Chinese University of Hong Kong, Shatin, New Territories, Hong Kong SAR, China

    Y M Dennis Lo, Rossa W K Chiu & Chunming Ding

  2. Department of Chemical Pathology, The Chinese University of Hong Kong, Shatin, New Territories, Hong Kong SAR, China

    Y M Dennis Lo, Nancy B Y Tsui, Rossa W K Chiu & Macy M S Heung

  3. Department of Obstetrics and Gynaecology, The Chinese University of Hong Kong, Shatin, New Territories, Hong Kong SAR, China

    Tze K Lau & Tse N Leung

  4. Harris Birthright Research Centre for Fetal Medicine, King's College Hospital, London, SE5 9RS, UK

    Ageliki Gerovassili & Kypros H Nicolaides

  5. Centre for Emerging Infectious Diseases, The Chinese University of Hong Kong, Shatin, New Territories, Hong Kong SAR, China

    Yongjie Jin & Chunming Ding

  6. Bioinformatics Program and Center for Advanced Biotechnology, Boston University, 36 Cummington Street, Boston, 02118, Massachusetts, USA

    Charles R Cantor

Authors
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  11. Chunming Ding
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Corresponding author

Correspondence to Y M Dennis Lo.

Ethics declarations

Competing interests

Y.M.D.L., N.B.Y.T., R.W.K.C., C.R.C. and C.D. have filed a patent application on the RNA-SNP allelic ratio technology described in this paper.

Y.M.D.L. has equity in Plasmagene Biosciences Limited.

C.R.C. and C.D. have equity in Sequenom Inc.

C.R.C. is the Chief Scientific Officer of Sequenom Inc.

Supplementary information

Supplementary Fig. 1

MS tracings of the PLAC4 SNP genotype in DNA samples of maternal blood cells and placentas and RNA samples of placentas and maternal plasma. (PDF 1523 kb)

Supplementary Table 1

Transcripts from chromosome 21 with high relative and absolute placental tissue expression as identified by the micorarray data mining strategy. (PDF 73 kb)

Supplementary Table 2

Sequences and allelic frequencies of four polymorphic SNPs located in the transcribed regions of PLAC4. (PDF 100 kb)

Supplementary Table 3

PLAC4 SNP genotype analysis using maternal blood cell and placental DNA as well as placental and maternal plasma RNA obtained from pregnant women carrying euploid fetuses. (PDF 73 kb)

Supplementary Table 4

Primer sequences of the RNA-SNP allelic ratio assay involving the PLAC4 SNP, rs8130833. (PDF 108 kb)

Supplementary Table 5

PCR primers for direct sequencing of PLAC4. (PDF 81 kb)

Supplementary Methods (PDF 125 kb)

Supplementary Note (PDF 74 kb)

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Lo, Y., Tsui, N., Chiu, R. et al. Plasma placental RNA allelic ratio permits noninvasive prenatal chromosomal aneuploidy detection. Nat Med 13, 218–223 (2007). https://doi.org/10.1038/nm1530

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