Assessment of free fetal DNA concentration in maternal plasma during the first trimester of pregnancy: comparative study between EDTA and PPT tubes - pilot study

J Matern Fetal Neonatal Med. 2015 Jan;28(2):172-6. doi: 10.3109/14767058.2014.908843. Epub 2014 May 14.

Abstract

Objective: To compare ethylenediamine tetraacetic acid (EDTA) tubes and plasma preparation tubes (PPT) for evaluating maternal plasma during the first trimester of pregnancy.

Methods: A cross-sectional study was conducted on 24 male fetuses in women between 6 and 14 weeks of pregnancy. Blood samples (10 mL) were collected and stored in EDTA and PPT tubes. Subsequently, the samples were centrifuged and sent for free fetal DNA extraction by means of the polymerase chain reaction (PCR) technique. The reactions were performed in a real time PCR machine for detecting the amplification products. The genome region chosen for performing the PCR reactions was a target specific for the Y chromosome, in which the DYS-14 marker was amplified only when the DNA was of male sex. The free fetal DNA concentration was given by the threshold cycle (TC). To compare the tubes, the paired Student t-test was used.

Results: The mean gestational age was 11.08 ± 2.30 weeks (range: 6-14). The mean TC for PPT was 30.08 ± 1.05 (range: 27.08-32.61) and for EDTA, 30.23 ± 0.96 (range: 28.01-32.09), but without statistical significance (p=0.357).

Conclusion: We did not observe any statistically significant difference in free fetal DNA concentration between the EDTA and PPT tubes.

Keywords: EDTA; PCR; PPT; first trimester of pregnancy; free fetal DNA.

Publication types

  • Comparative Study
  • Evaluation Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Anticoagulants / pharmacology*
  • Blood Preservation / methods*
  • Blood Specimen Collection / methods*
  • Cell Cycle Proteins / analysis
  • Cell Cycle Proteins / genetics
  • Cross-Sectional Studies
  • DNA / analysis
  • DNA / blood*
  • Edetic Acid / pharmacology*
  • Female
  • Fetus / metabolism*
  • Gestational Age
  • Humans
  • Male
  • Pilot Projects
  • Polymerase Chain Reaction
  • Pregnancy
  • Pregnancy Trimester, First / blood*
  • Pregnancy Trimester, First / genetics

Substances

  • Anticoagulants
  • Cell Cycle Proteins
  • TSPY1 protein, human
  • DNA
  • Edetic Acid