A Universal Droplet Digital PCR Approach for Monitoring of Graft Health After Transplantation Using a Preselected SNP Set

Julia Beck; Michael Oellerich; Ekkehard Schütz

doi:10.1007/978-1-4939-7778-9_19

A Universal Droplet Digital PCR Approach for Monitoring of Graft Health After Transplantation Using a Preselected SNP Set

Methods Mol Biol. 2018;1768:335-348. doi: 10.1007/978-1-4939-7778-9_19.

Authors

Julia Beck¹, Michael Oellerich², Ekkehard Schütz³

Affiliations

¹ Chronix Biomedical GmbH, Göttingen, Germany. jbeck@chronixbiomedical.de.
² Institute for Clinical Pharmacology, University Medical Center Göttingen, Göttingen, Germany.
³ Chronix Biomedical GmbH, Göttingen, Germany.

PMID: 29717452
DOI: 10.1007/978-1-4939-7778-9_19

Abstract

Transplanted organs release cell-free DNA into the bloodstream of the recipient. This graft-derived cell-free DNA (GcfDNA) is a sensitive biomarker for organ health, since higher GcfDNA levels are indicative of increased cell-death in the graft. This protocol describes a method to measure relative GcfDNA concentrations by ddPCR assays. The method uses a set of preselected SNP assays from which the informative SNPs for each recipient-donor combination are selected in a straightforward two-step procedure that requires only one blood draw. Sampling of donor tissue and separate genotyping is not required, rendering the technique applicable also to patients, whose transplantation was not recent. In these patients there will be mostly no access to donor DNA anymore.

Keywords: Rejection monitoring; Transplantation.

MeSH terms

Biomarkers / blood
Cell-Free Nucleic Acids / genetics
Cell-Free Nucleic Acids / isolation & purification*
Graft Rejection / blood
Graft Rejection / diagnosis*
Graft Rejection / genetics
Humans
Organ Transplantation / adverse effects*
Polymerase Chain Reaction / instrumentation
Polymerase Chain Reaction / methods*
Polymorphism, Single Nucleotide*

Substances

Biomarkers
Cell-Free Nucleic Acids