Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Technical Report
  • Published:

Plasma placental RNA allelic ratio permits noninvasive prenatal chromosomal aneuploidy detection

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.

This is a preview of subscription content, access via your institution

Access options

Rent or buy this article

Prices vary by article type

from$1.95

to$39.95

Prices may be subject to local taxes which are calculated during checkout

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.

Similar content being viewed by others

References

  1. Alfirevic, Z., Sundberg, K. & Brigham, S. Amniocentesis and chorionic villus sampling for prenatal diagnosis. Cochrane Database Syst. Rev. CD003252 (2003).

  2. Malone, F.D. et al. First-trimester or second-trimester screening, or both, for Down's syndrome. N. Engl. J. Med. 353, 2001–2011 (2005).

    Article  CAS  PubMed  Google Scholar 

  3. Wapner, R. et al. First-trimester screening for trisomies 21 and 18. N. Engl. J. Med. 349, 1405–1413 (2003).

    Article  CAS  PubMed  Google Scholar 

  4. Bianchi, D.W., Flint, A.F., Pizzimenti, M.F., Knoll, J.H. & Latt, S.A. Isolation of fetal DNA from nucleated erythrocytes in maternal blood. Proc. Natl. Acad. Sci. USA 87, 3279–3283 (1990).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  5. Cheung, M.C., Goldberg, J.D. & Kan, Y.W. Prenatal diagnosis of sickle cell anaemia and thalassaemia by analysis of fetal cells in maternal blood. Nat. Genet. 14, 264–268 (1996).

    Article  CAS  PubMed  Google Scholar 

  6. Bianchi, D.W. et al. PCR quantitation of fetal cells in maternal blood in normal and aneuploid pregnancies. Am. J. Hum. Genet. 61, 822–829 (1997).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  7. Bianchi, D.W. et al. Fetal gender and aneuploidy detection using fetal cells in maternal blood: analysis of NIFTY I data. National Institute of Child Health and Development Fetal Cell Isolation Study. Prenat. Diagn. 22, 609–615 (2002).

    Article  CAS  PubMed  Google Scholar 

  8. Lo, Y.M.D. et al. Presence of fetal DNA in maternal plasma and serum. Lancet 350, 485–487 (1997).

    Article  CAS  PubMed  Google Scholar 

  9. Lo, Y.M.D. et al. Quantitative analysis of fetal DNA in maternal plasma and serum: implications for noninvasive prenatal diagnosis. Am. J. Hum. Genet. 62, 768–775 (1998).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  10. Kaiser, J. Prenatal diagnosis. An earlier look at baby's genes. Science 309, 1476–1478 (2005).

    Article  CAS  PubMed  Google Scholar 

  11. Lo, Y.M.D. et al. Increased fetal DNA concentrations in the plasma of pregnant women carrying fetuses with trisomy 21. Clin. Chem. 45, 1747–1751 (1999).

    CAS  PubMed  Google Scholar 

  12. Lee, T. et al. Down syndrome and cell-free fetal DNA in archived maternal serum. Am. J. Obstet. Gynecol. 187, 1217–1221 (2002).

    Article  PubMed  Google Scholar 

  13. Farina, A. et al. Evaluation of cell-free fetal DNA as a second-trimester maternal serum marker of Down syndrome pregnancy. Clin. Chem. 49, 239–242 (2003).

    Article  CAS  PubMed  Google Scholar 

  14. Ng, E.K.O. et al. mRNA of placental origin is readily detectable in maternal plasma. Proc. Natl. Acad. Sci. USA 100, 4748–4753 (2003).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  15. Ferguson-Smith, M.A. Placental mRNA in maternal plasma: prospects for fetal screening. Proc. Natl. Acad. Sci. USA 100, 4360–4362 (2003).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  16. Oudejans, C.B. et al. Detection of chromosome 21-encoded mRNA of placental origin in maternal plasma. Clin. Chem. 49, 1445–1449 (2003).

    Article  CAS  PubMed  Google Scholar 

  17. Tsui, N.B.Y. et al. Systematic micro-array based identification of placental mRNA in maternal plasma: towards non-invasive prenatal gene expression profiling. J. Med. Genet. 41, 461–467 (2004).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  18. Chim, S.S.C. et al. Detection of the placental epigenetic signature of the maspin gene in maternal plasma. Proc. Natl. Acad. Sci. USA 102, 14753–14758 (2005).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  19. Lui, Y.Y.N. et al. Predominant hematopoietic origin of cell-free DNA in plasma and serum after sex-mismatched bone marrow transplantation. Clin. Chem. 48, 421–427 (2002).

    CAS  PubMed  Google Scholar 

  20. Kido, S. et al. D21S418E identifies a cAMP-regulated gene located on chromosome 21q22.3 that is expressed in placental syncytiotrophoblast and choriocarcinoma cells. Genomics 17, 256–259 (1993).

    Article  CAS  PubMed  Google Scholar 

  21. Ding, C., Maier, E., Roscher, A.A., Braun, A. & Cantor, C.R. Simultaneous quantitative and allele-specific expression analysis with real competitive PCR. BMC Genet. 5, 8 (2004).

    Article  PubMed  PubMed Central  Google Scholar 

  22. Wapner, R.J. Invasive prenatal diagnostic techniques. Semin. Perinatol. 29, 401–404 (2005).

    Article  PubMed  Google Scholar 

  23. Amor, D.J. et al. Health and developmental outcome of children following prenatal diagnosis of confined placental mosaicism. Prenat. Diagn. 26, 443–448 (2006).

    Article  PubMed  Google Scholar 

  24. Anker, P., Mulcahy, H. & Stroun, M. Circulating nucleic acids in plasma and serum as a noninvasive investigation for cancer: time for large-scale clinical studies? Int. J. Cancer 103, 149–152 (2003).

    Article  CAS  PubMed  Google Scholar 

  25. Jurinke, C., Oeth, P. & van den Boom, D. MALDI-TOF mass spectrometry: a versatile tool for high-performance DNA analysis. Mol. Biotechnol. 26, 147–164 (2004).

    Article  CAS  PubMed  Google Scholar 

Download references

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

Authors and Affiliations

Authors

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)

Rights and permissions

Reprints and permissions

About this article

Cite this article

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

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/nm1530

This article is cited by

Search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing