Plasma donor-derived cell-free DNA kinetics after kidney transplantation using a single tube multiplex PCR assay

PLoS One. 2018 Dec 6;13(12):e0208207. doi: 10.1371/journal.pone.0208207. eCollection 2018.

Abstract

Background: After transplantation, cell-free DNA derived from the donor organ (ddcfDNA) can be detected in the recipient's circulation. We aimed to quantify ddcfDNA levels in plasma of kidney transplant recipients thereby investigating the kinetics of this biomarker after transplantation and determining biological variables that influence ddcfDNA kinetics in stable and non-stable patients.

Materials and methods: From 107 kidney transplant recipients, plasma samples were collected longitudinally after transplantation (day 1-3 months) within a multicenter set-up. Cell-free DNA from the donor was quantified in plasma as a fraction of the total cell-free DNA by next generation sequencing using a targeted, multiplex PCR-based method for the analysis of single nucleotide polymorphisms. A subgroup of stable renal transplant recipients was identified to determine a ddcfDNA threshold value.

Results: In stable transplant recipients, plasma ddcfDNA% decreased to a mean (SD) ddcfDNA% of 0.46% (± 0.21%) which was reached 9.85 (± 5.6) days after transplantation. A ddcfDNA threshold value of 0.88% (mean + 2SD) was determined in kidney transplant recipients. Recipients that did not reach this threshold ddcfDNA value within 10 days after transplantation showed a higher ddcfDNA% on the first day after transplantation and demonstrated a higher individual baseline ddcfDNA%.

Conclusion: In conclusion, plasma ddcfDNA fractions decreased exponentially within 10 days after transplantation to a ddcfDNA threshold value of 0.88% or less. To investigate the role of ddcfDNA for rejection monitoring of the graft, future research is needed to determine causes of ddcfDNA% increases above this threshold value.

Publication types

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

MeSH terms

  • Blood Donors
  • Cell-Free Nucleic Acids / blood*
  • Humans
  • Kidney Transplantation / methods*
  • Kinetics
  • Longitudinal Studies
  • Multiplex Polymerase Chain Reaction / methods*
  • Polymorphism, Single Nucleotide / genetics
  • Prospective Studies
  • Transplant Recipients

Substances

  • Cell-Free Nucleic Acids

Grants and funding

This work was supported by PhD Fellowship of the Research Foundation – Flanders (FWO) was granted to EG (1136317N), http://www.fwo.be/nl/, to EG. This work was funded by Flanders Innovation and Entrepreneurship (140574) granted to JDF, DA, BDW. https://www.vlaio.be/nl, to JDF, DA, BDW. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.