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Technical Briefs |
1
Molecular Biology Laboratory, Department of Clinical Chemistry and
2
Department of Obstetrics and Gynecology, University Hospital ‘Vrije Universiteit’, 1007 MB Amsterdam, The Netherlands;
a address correspondence to this author at: Molecular Biology Laboratory, Department of Clinical Chemistry, University Hospital ‘Vrije Universiteit’, PO Box 7057, 1007 MB Amsterdam, The Netherlands
Fetal DNA is present in plasma of pregnant women in amounts almost 1000-fold higher than the DNA present in intact fetal cells circulating in maternal peripheral blood (1)(2). The clinical usefulness of fetal DNA from plasma for noninvasive prenatal diagnosis has been demonstrated recently through analysis of the fetal RhD status by PCR-based approaches (3)(4)(5). We explored the possibility that not all of this fetal DNA is soluble and cell-free, but that part of it is still cell-associated.
Heparin blood samples of 25–30 mL were obtained after informed consent
from 38 pregnant women attending the Prenatal Diagnostic Center at week
7–16 of gestation. Several weeks after blood withdrawal, chorionic
villus sampling or amniocentesis was performed as planned at
each patient’s first visit. The protocol was approved by the Committee
of Medical Ethics of the University Hospital ‘Vrije Universiteit’.
Blood samples were processed on the day of withdrawal and were
centrifuged on a discontinuous Percoll gradient as described previously
(6)(7). In brief, after dilution of the heparin
blood in Hanks’ balanced salt solution; 1:2 dilution (Life
Technologies), blood was layered on a 5-step Percoll density gradient
consisting of 15 mL of 600 mL/L and 5 mL each of 550, 500, 450,
and 400 mL/L Percoll in Hanks’ balanced salt solution. After
centrifugation for 25 min at 1000g at room
temperature, plasma samples were removed
Acknowledgments
Footnotes
References
The following articles in journals at HighWire Press have cited this article:
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  T.V. Zolotukhina, N.V. Shilova, and E. Y. Voskoboeva Analysis of Cell-free Fetal DNA in Plasma and Serum of Pregnant Women J. Histochem. Cytochem., March 1, 2005; 53(3): 297 - 299. [Abstract] [Full Text] [PDF]
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 T. H. Rainer, N. Y.L. Lam, N. B.Y. Tsui, E. K.O. Ng, R. W.K. Chiu, G. M. Joynt, and Y.M. D. Lo Effects of Filtration on Glyceraldehyde-3-Phosphate Dehydrogenase mRNA in the Plasma of Trauma Patients and Healthy Individuals Clin. Chem., January 1, 2004; 50(1): 206 - 208. [Full Text] [PDF]
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S. Hristoskova, W. Holzgreve, and S. Hahn Fetal Nucleated Erythrocytes in Maternal Circulation Do Not Display a Classic Membrane-associated Apoptotic Characteristic (Phosphatidylserine Exposure) Despite Being Positive by Terminal dUTP Nuclear End Labeling Clin. Chem., November 1, 2003; 49(11): 1934 - 1937. [Full Text] [PDF]
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 J. Guibert, A. Benachi, A.-G. Grebille, P. Ernault, J.-R. Zorn, and J.-M. Costa Kinetics of SRY gene appearance in maternal serum: detection by real time PCR in early pregnancy after assisted reproductive technique Hum. Reprod., August 1, 2003; 18(8): 1733 - 1736. [Abstract] [Full Text] [PDF]
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 E. K. O. Ng, N. B. Y. Tsui, T. K. Lau, T. N. Leung, R. W. K. Chiu, N. S. Panesar, L. C. W. Lit, K.-W. Chan, and Y. M. D. Lo From the Cover: mRNA of placental origin is readily detectable in maternal plasma PNAS, April 15, 2003; 100(8): 4748 - 4753. [Abstract] [Full Text] [PDF]
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R. M. Angert, E. S. LeShane, Y.M. D. Lo, L. Y.S. Chan, L. C. Delli-Bovi, and D. W. Bianchi Fetal Cell-free Plasma DNA Concentrations in Maternal Blood Are Stable 24 Hours after Collection: Analysis of First- and Third-Trimester Samples Clin. Chem., January 1, 2003; 49(1): 195 - 198. [Full Text] [PDF]
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 N. C. Lambert, Y. M. D. Lo, T. D. Erickson, T. S. Tylee, K. A. Guthrie, D. E. Furst, and J. L. Nelson Male microchimerism in healthy women and women with scleroderma: cells or circulating DNA? A quantitative answer Blood, September 26, 2002; 100(8): 2845 - 2851. [Abstract] [Full Text] [PDF]
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 X. Y. Zhong, W. Holzgreve, and S. Hahn Cell-free fetal DNA in the maternal circulation does not stem from the transplacental passage of fetal erythroblasts Mol. Hum. Reprod., September 1, 2002; 8(9): 864 - 870. [Abstract] [Full Text] [PDF]
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E. K.O. Ng, N. B.Y. Tsui, N. Y.L. Lam, R. W.K. Chiu, S. C.H. Yu, S.C. C. Wong, E. S.F. Lo, T. H. Rainer, P. J. Johnson, and Y.M. D. Lo Presence of Filterable and Nonfilterable mRNA in the Plasma of Cancer Patients and Healthy Individuals Clin. Chem., August 1, 2002; 48(8): 1212 - 1217. [Abstract] [Full Text] [PDF]
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 G. Vona, C. Beroud, A. Benachi, A. Quenette, J. P. Bonnefont, S. Romana, Y. Dumez, B. Lacour, and P. Paterlini-Brechot Enrichment, Immunomorphological, and Genetic Characterization of Fetal Cells Circulating in Maternal Blood Am. J. Pathol., January 1, 2002; 160(1): 51 - 58. [Abstract] [Full Text] [PDF]
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D. W. Bianchi, E. S. LeShane, and J. M. Cowan Large Amounts of Cell-free Fetal DNA Are Present in Amniotic Fluid Clin. Chem., October 1, 2001; 47(10): 1867 - 1869. [Full Text] [PDF]
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S. Hristoskova, W. Holzgreve, S. Hahn, I. J. van Wijk, and C. B.M. Oudejans More Than One-Half of the Erythroblasts in the Fetal Circulation and Cord Blood Are TUNEL Positive Drs. van Wijk and Oudejans respond: Clin. Chem., October 1, 2001; 47(10): 1870 - 1871. [Full Text] [PDF]
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R. W. K. Chiu, L. L. M. Poon, T. K. Lau, T. N. Leung, E. M. C. Wong, and Y. M. D. Lo Effects of Blood-Processing Protocols on Fetal and Total DNA Quantification in Maternal Plasma Clin. Chem., September 1, 2001; 47(9): 1607 - 1613. [Abstract] [Full Text] [PDF]
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Y.M. D. Lo Fetal DNA in Maternal Plasma: Biology and Diagnostic Applications Clin. Chem., December 1, 2000; 46(12): 1903 - 1906. [Abstract] [Full Text] [PDF]
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L. L.M. Poon, T. N. Leung, T. K. Lau, and Y.M. D. Lo Presence of Fetal RNA in Maternal Plasma Clin. Chem., November 1, 2000; 46(11): 1832 - 1834. [Full Text] [PDF]
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Y.M. D. Lo, T. K. Lau, L. Y.S. Chan, T. N. Leung, and A. M.Z. Chang Quantitative Analysis of the Bidirectional Fetomaternal Transfer of Nucleated Cells and Plasma DNA Clin. Chem., September 1, 2000; 46(9): 1301 - 1309. [Abstract] [Full Text] [PDF]
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