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Clin Chem. 2005 Sep;51(9):1598-604. Epub 2005 Jul 14.

Optimized real-time quantitative PCR measurement of male fetal DNA in maternal plasma.

Author information

  • 1University Women's Hospital/Department of Research, University Hospital Basel, Basel, Switzerland. Bernhard.Zimmermann@uwe.ac.uk

Abstract

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.

PMID:
16020496
[PubMed - indexed for MEDLINE]
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