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A fast and simple method for detecting and quantifying donor-derived cell-free DNA in sera of solid organ transplant recipients as a biomarker for graft function

  • Martina Adamek , Gerhard Opelz , Katrin Klein , Christian Morath and Thuong Hien Tran EMAIL logo
Published/Copyright: November 17, 2015
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Clinical Chemistry and Laboratory Medicine (CCLM)
From the journal Clinical Chemistry and Laboratory Medicine (CCLM) Volume 54 Issue 7

Abstract

Background: Timely detection of graft rejection is an important issue in the follow-up care after solid organ transplantation. Until now, biopsy has been considered the “gold standard” in the diagnosis of graft rejection. However, non-invasive tests such as monitoring the levels of cell-free DNA (cfDNA) as a sensitive biomarker for graft integrity have attracted increasing interest. The rationale of this approach is that a rejected organ will lead to a significant release of donor-derived cfDNA, which can be detected in the serum of the transplant recipient.

Methods: We have developed a novel quantitative real-time PCR (qPCR) approach for detecting an increase of donor-derived cfDNA in the recipient’s serum. Common insertion/deletion (InDel) genetic polymorphisms, which differ between donor and recipient, are targeted in our qPCR assay. In contrast to some other strategies, no specific donor/recipient constellations such as certain gender combinations or human leukocyte antigen (HLA) discrepancies are required for the application of our test.

Results: The method was first validated with serial dilutions of serum mixtures obtained from healthy blood donors and then used to determine donor-derived cfDNA levels in patients’ sera within the first 3 days after their kidney transplantation had been performed.

Conclusions: Our method represents a universally applicable, simple and cost-effective tool which can potentially be used to detect graft dysfunction in transplant recipients.

Keywords: biomarker; cell-free DNA; graft rejection; InDel; kidney transplantation; qPCR
Corresponding author: Thuong Hien Tran, MD, Transplantation Immunology, Institute of Immunology, University of Heidelberg, Im Neuenheimer Feld 305, 69120 Heidelberg, Germany, E-mail:

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Supplemental Material:

The online version of this article (DOI: 10.1515/cclm-2015-0622) offers supplementary material, available to authorized users.

Received: 2015-7-1
Accepted: 2015-10-17
Published Online: 2015-11-17
Published in Print: 2016-7-1

©2016 by De Gruyter

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  5. The quality indicator paradox
  6. Review
  7. Clinical utility of the (-2)proPSA and evaluation of the evidence: a systematic review
  8. Mini Review
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  10. EFLM Position Paper
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https://doi.org/10.1515/cclm-2015-0622
Keywords for this article
biomarker; cell-free DNA; graft rejection; InDel; kidney transplantation; qPCR

Articles in the same Issue

  1. Frontmatter
  2. In Memoriam: Gérard Siest (1936–2016)
  3. Editorials
  4. Improving diagnosis and reducing diagnostic errors: the next frontier of laboratory medicine
  5. The quality indicator paradox
  6. Review
  7. Clinical utility of the (-2)proPSA and evaluation of the evidence: a systematic review
  8. Mini Review
  9. Why are clinical practice guidelines not followed?
  10. EFLM Position Paper
  11. Patient identification and tube labelling – a call for harmonisation
  12. Genetics and Molecular Diagnostics
  13. A fast and simple method for detecting and quantifying donor-derived cell-free DNA in sera of solid organ transplant recipients as a biomarker for graft function
  14. Performance of two commercially available BCR-ABL1 quantification assays that use an international reporting scale
  15. HAND1 loss-of-function mutation associated with familial dilated cardiomyopathy
  16. General Clinical Chemistry and Laboratory Medicine
  17. Performance criteria and quality indicators for the post-analytical phase
  18. Assessing the commutability of reference material formats for the harmonization of amyloid-β measurements
  19. Quantitative determination of four immunosuppressants by high resolution mass spectrometry (HRMS)
  20. Measurement of plasma vitamin K1 (phylloquinone) and K2 (menaquinones-4 and -7) using HPLC-tandem mass spectrometry
  21. Therapeutic drug monitoring of infliximab: performance evaluation of three commercial ELISA kits
  22. High level of oxysterols in neonatal cholestasis: a pitfall in analysis of biochemical markers for Niemann-Pick type C disease
  23. Reference Values and Biological Variations
  24. Reference intervals of plasma homoarginine from the German Gutenberg Health Study
  25. TSH and fT4 during pregnancy: an observational study and a review of the literature
  26. Cancer Diagnostics
  27. Mean corpuscular volume levels and all-cause and liver cancer mortality
  28. Cardiovascular Diseases
  29. Circulating endothelial-derived apoptotic microparticles and insulin resistance in non-diabetic patients with chronic heart failure
  30. Hematology and Coagulation
  31. Platelet aggregation in response to ADP is highly variable in normal donors and patients on anti-platelet medication
  32. Letters to the Editor
  33. EQA-derived metrics to assess overall instrument performance
  34. Misidentification in laboratory medicine and diagnostic process: a neglected problem calling for action
  35. Impact of the routine implementation of automated indirect immunofluorescence antinuclear antibody analysis: 1 year of experience
  36. The importance of angiogenic markers in the differential diagnosis of HELLP syndrome vs. non-HELLP thrombocytopenia
  37. Bisalbuminemia accompanying bisalbuminuria detected in capillary electrophoresis, not in gel electrophoresis
  38. Prognostic value of red blood cell distribution width in acute pancreatitis patients admitted to intensive care units: an analysis of a publicly accessible clinical database MIMIC II
  39. Differences in analytical and biological results between older and newer lots of a widely used irisin immunoassay question the validity of previous studies
  40. A diagnostic algorithm for the detection of inhibitors against coagulation Factor V
  41. Interference of anticoagulants on coagulation testing
  42. Congress Abstracts
  43. Swiss MedLab 2016 and 74th Annual Meeting of the Swiss Society of Microbiology SSM, Bern, 13–16 June 2016
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