The current standard for graft rejection surveillance is endomyocardial biopsy (EMB), an invasive procedure with rare but potentially serious complications. Detection of circulating donor-derived cell-free DNA (ddcfDNA) is an option for noninvasive monitoring of graft injury and rejection. A 63-year-old man and a 65-year-old woman were monitored by EMB for allograft rejection. A total of 48 single-nucleotide polymorphisms with a minor allele frequency range of 0.4-0.5 were screened to distinguish donor and recipient DNA based on homozygosity, and digital droplet PCR was used to analyze ddcfDNA concentrations. Both subjects suffered rejection within the first 6 months after transplantation. The maximal ddcfDNA level of 270 copies (cp)/ml during EMB-confirmed acute cellular rejection (ACR; mild grade 1R/2, patient 1), and the maximal concentration of 1,846 cp/ml in the case of EMB-confirmed antibody-mediated rejection (AMR; grade 1+; patient 2), was detected. Individual monitoring of ddcfDNA dynamics from the 1st to the 6th month posttransplant reflected cardiac graft injury in patients suffering ACR or AMR, meaning that ddcfDNA may serve as a noninvasive biomarker.

3rd Department of Internal Medicine 1st Faculty of Medicine Charles University 12808 Prague Czech Republic

Cardio Center Institute for Clinical and Experimental Medicine 14021 Prague Czech Republic

Center for Experimental Medicine Institute for Clinical and Experimental Medicine 14021 Prague Czech Republic

Department of Anthropology and Human Genetics Faculty of Science Charles University 12808 Prague Czech Republic

Laboratory of Gene Expression Institute of Biotechnology Czech Academy of Science BIOCEV 25250 Vestec Czech Republic

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