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Received on March 22, 2005
Accepted on June 23, 2005
Molecular Diagnostics and Genetics |
1 University Women's Hospital/Department of Research, University Hospital Basel, Switzerland
2 Harris Birthright Research Centre for Fetal Medicine, King's College Hospital, London, United Kingdom
* To whom correspondence should be addressed. E-mail: Bernhard.Zimmermann{at}uwe.ac.uk.
Background: Circulating fetal DNA (cfDNA) in maternal plasma has been measured to investigate its possible relationship with pregnancy-related disorders, including fetal trisomy 21 and preeclampsia. The circulating concentrations of single-copy fetal genes, however, are close to the detection limits of PCR methods.
Methods: We optimized a protocol for the real-time quantitative PCR amplification of the multicopy sequence DYS14 on the Y-chromosome. This was compared with an established real-time PCR assay for the single-copy SRY gene.
Results: By probit regression analysis, the measurements of male DNA by the DYS14 assay had a 10-fold lower detection limit (0.4 genome equivalents) than did measurements of SRY. For plasma samples from women in the first trimester of pregnancy, imprecision (CV) was 2%-22% when amplifying DYS14 compared with 26%-140% for SRY.
Conclusions: The low copy numbers of fetal DNA in plasma of women in the first trimester of pregnancy cannot be measured precisely when targeting single-copy sequences. Better results are obtained by amplifying a sequence that is present in multiple copies per male genome.
The following articles in journals at HighWire Press have cited this article:
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  O. Lapaire, T. Volgmann, S. Grill, I. Hosli, R. Zanetti-Daellenbach, X. Y. Zhong, and W. Holzgreve Significant Correlation Between Maternal Body Mass Index at Delivery and in the Second Trimester, and Second Trimester Circulating Total Cell-free DNA Levels Reproductive Sciences, March 1, 2009; 16(3): 274 - 279. [Abstract] [PDF]
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 C. F. Wright and H. Burton The use of cell-free fetal nucleic acids in maternal blood for non-invasive prenatal diagnosis Hum. Reprod. Update, January 1, 2009; 15(1): 139 - 151. [Abstract] [Full Text] [PDF]
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 F. M. F. Lun, R. W. K. Chiu, K. C. Allen Chan, T. Yeung Leung, T. Kin Lau, and Y. M. Dennis Lo Microfluidics Digital PCR Reveals a Higher than Expected Fraction of Fetal DNA in Maternal Plasma Clin. Chem., October 1, 2008; 54(10): 1664 - 1672. [Abstract] [Full Text] [PDF]
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 A. Yin, E.H.Y. Ng, X. Zhang, Y. He, J. Wu, and K.Y. Leung Correlation of maternal plasma total cell-free DNA and fetal DNA levels with short term outcome of first-trimester vaginal bleeding Hum. Reprod., June 1, 2007; 22(6): 1736 - 1743. [Abstract] [Full Text] [PDF]
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 I. Stanghellini, R. Bertorelli, L. Capone, V. Mazza, C. Neri, A. Percesepe, and A. Forabosco Quantitation of fetal DNA in maternal serum during the first trimester of pregnancy by the use of a DAZ repetitive probe Mol. Hum. Reprod., September 1, 2006; 12(9): 587 - 591. [Abstract] [Full Text] [PDF]
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D. J. Huang, B. G. Zimmermann, W. Holzgreve, and S. Hahn Use of an Automated Method Improves the Yield and Quality of Cell-Free Fetal DNA Extracted from Maternal Plasma Clin. Chem., December 1, 2005; 51(12): 2419 - 2420. [Full Text] [PDF]
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