Authors:	Ehlert, Tobias Tug, Suzan Brahmer, Alexandra Neef, Vanessa Heid, Florian Werner, Christian Jansen-Winkeln, Boris Kneist, Werner Lang, Hauke Gockel, Ines Simon, Perikles
Title:	Establishing PNB-qPCR for quantifying minimal ctDNA concentrations during tumour resection
Online publication date:	12-Oct-2022
Year of first publication:	2017
Language:	english
Abstract:	The analysis of blood plasma or serum as a non-invasive alternative to tissue biopsies is a much-pursued goal in cancer research. Various methods and approaches have been presented to determine a patient’s tumour status, chances of survival, and response to therapy from serum or plasma samples. We established PNB-qPCR (Pooled, Nested, WT-Blocking qPCR), a highly specific nested qPCR with various modifications to detect and quantify minute amounts of circulating tumour DNA (ctDNA) from very limited blood plasma samples. PNB-qPCR is a nested qPCR technique combining ARMS primers, blocking primers, LNA probes, and pooling of multiple first round products for sensitive quantification of the seven most frequent point mutations in KRAS exon 2. Using this approach, we were able to characterize ctDNA and total cell-free DNA (cfDNA) kinetics by selective amplification of KRAS mutated DNA fragments in the blood plasma over the course of tumour resection and the surrounding days. Whereas total cfDNA concentrations increased over the surgical and regenerative process, ctDNA levels showed a different scheme, rising only directly after tumour resection and about three days after the surgery. For the first time, we present insights into the impact of surgery on the release of ctDNA and total cfDNA.
DDC:	610 Medizin 610 Medical sciences
Institution:	Johannes Gutenberg-Universität Mainz
Department:	FB 02 Sozialwiss., Medien u. Sport FB 04 Medizin
Place:	Mainz
ROR:	https://ror.org/023b0x485
DOI:	http://doi.org/10.25358/openscience-7945
Version:	Published version
Publication type:	Zeitschriftenaufsatz
License:	CC BY
Information on rights of use:	https://creativecommons.org/licenses/by/4.0/
Journal:	Scientific reports 7
Pages or article number:	Art. 8876
Publisher:	Macmillan Publishers Limited, part of Springer Nature
Publisher place:	London
Issue date:	2017
ISSN:	2045-2322
Publisher URL:	http://dx.doi.org/10.1038/s41598-017-09137-w
Publisher DOI:	10.1038/s41598-017-09137-w
Appears in collections:	DFG-OA-Publizieren (2012 - 2017)