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Title:Feasibility of droplet digital PCR analysis of plasma cell-free DNA from kidney transplant patients
Authors:ID Jerič Kokelj, Barbara (Author)
ID Štalekar, Maja (Author)
ID Vencken, Sebastian (Author)
ID Dobnik, David (Author)
ID Kogovšek, Polona (Author)
ID Stanonik, Matjaž (Author)
ID Arnol, Miha (Author)
ID Ravnikar, Maja (Author)
Files:URL URL - Source URL, visit https://www.frontiersin.org/articles/10.3389/fmed.2021.748668/full
 
.pdf PDF - Presentation file, download (842,05 KB)
MD5: 7FE0006D58E14DFED953376D087E1AAB
 
Language:English
Typology:1.01 - Original Scientific Article
Organization:Logo NIB - National Institute of Biology
Abstract:Increasing research demonstrates the potential of donor-derived cell-free DNA (dd-cfDNA) as a biomarker for monitoring the health of various solid organ transplants. Several methods have been proposed for cfDNA analysis, including real-time PCR, digital PCR, and next generation sequencing-based approaches. We sought to revise the droplet digital PCR (ddPCR)-based approach to quantify relative dd-cfDNA in plasma from kidney transplant (KTx) patients using a novel pilot set of assays targeting single nucleotide polymorphisms that have a very high potential to distinguish cfDNA from two individuals. The assays are capable of accurate quantification of down to 0.1% minor allele content when analyzing 165 ng of human DNA. We found no significant differences in the yield of extracted cfDNA using the three different commercial kits tested. More cfDNA was extracted from the plasma of KTx patients than from healthy volunteers, especially early after transplantation. The median level of donor-derived minor alleles in KTx samples was 0.35%. We found that ddPCR using the evaluated assays within specific range is suitable for analysis of KTx patientsʼ plasma but recommend prior genotyping of donor DNA and performing reliable preamplification of cfDNA.
Keywords:kidney transplantation, droplet digital PCR, plasma cell-free DNA, minor allele quantification, assay evaluation, graft health monitoring
Publication status:Published
Publication version:Version of Record
Publication date:08.10.2021
Year of publishing:2021
Number of pages:str. 1-9
Numbering:Vol. 8
PID:20.500.12556/DiRROS-19480 New window
UDC:616.6:615
ISSN on article:2296-858X
DOI:10.3389/fmed.2021.748668 New window
COBISS.SI-ID:79997955 New window
Note:Soavtorji: Maja Štalekar, Sebastian Vencken, David Dobnik, Polona Kogovšek, Matjaž Stanonik, Miha Arnol and Maja Ravnikar; Nasl. z nasl. zaslona; Opis vira z dne 11. 10. 2021; Št. članka: 748668;
Publication date in DiRROS:19.07.2024
Views:683
Downloads:266
Metadata:XML DC-XML DC-RDF
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JERIČ KOKELJ, Barbara, ŠTALEKAR, Maja, VENCKEN, Sebastian, DOBNIK, David, KOGOVŠEK, Polona, STANONIK, Matjaž, ARNOL, Miha and RAVNIKAR, Maja, 2021, Feasibility of droplet digital PCR analysis of plasma cell-free DNA from kidney transplant patients. Frontiers in medicine [online]. 2021. Vol. 8, p. 1–9. [Accessed 4 April 2025]. DOI 10.3389/fmed.2021.748668. Retrieved from: https://dirros.openscience.si/IzpisGradiva.php?lang=eng&id=19480
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Record is a part of a journal

Title:Frontiers in medicine
Shortened title:Front. med.
Publisher:Frontiers Media S.A.
ISSN:2296-858X
COBISS.SI-ID:523095065 New window

Document is financed by a project

Funder:ARIS - Slovenian Research and Innovation Agency
Project number:P4-0165-2015
Name:Biotehnologija in sistemska biologija rastlin
Funder:ARIS - Slovenian Research and Innovation Agency
Project number:P4-0407-2019
Name:Okoljska in aplikativna virologija: virusi, prijatelji in sovražniki
Funder:Other - Other funder or multiple funders
Funding programme:Ministry of Education, Science and Sport of the Republic of Slovenia
Project number:C3330-17-529023
Name:Raziskovalci-2.0-NIB-529023

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License:CC BY 4.0, Creative Commons Attribution 4.0 International
Link:http://creativecommons.org/licenses/by/4.0/
Description:This is the standard Creative Commons license that gives others maximum freedom to do what they want with the work as long as they credit the author.

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