Establishing PNB-qPCR for quantifying minimal ctDNA concentrations during tumour resection

dc.contributor.authorEhlert, Tobias
dc.contributor.authorTug, Suzan
dc.contributor.authorBrahmer, Alexandra
dc.contributor.authorNeef, Vanessa
dc.contributor.authorHeid, Florian
dc.contributor.authorWerner, Christian
dc.contributor.authorJansen-Winkeln, Boris
dc.contributor.authorKneist, Werner
dc.contributor.authorLang, Hauke
dc.contributor.authorGockel, Ines
dc.contributor.authorSimon, Perikles
dc.date.accessioned2022-10-12T09:56:13Z
dc.date.available2022-10-12T09:56:13Z
dc.date.issued2017
dc.description.abstractThe 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.en_GB
dc.description.sponsorshipDFG, Open Access-Publizieren Universität Mainz / Universitätsmedizinde
dc.identifier.doihttp://doi.org/10.25358/openscience-7945
dc.identifier.urihttps://openscience.ub.uni-mainz.de/handle/20.500.12030/7960
dc.language.isoengde
dc.rightsCC-BY-4.0*
dc.rights.urihttps://creativecommons.org/licenses/by/4.0/*
dc.subject.ddc610 Medizinde_DE
dc.subject.ddc610 Medical sciencesen_GB
dc.titleEstablishing PNB-qPCR for quantifying minimal ctDNA concentrations during tumour resectionen_GB
dc.typeZeitschriftenaufsatzde
jgu.journal.titleScientific reportsde
jgu.journal.volume7de
jgu.organisation.departmentFB 02 Sozialwiss., Medien u. Sportde
jgu.organisation.departmentFB 04 Medizinde
jgu.organisation.nameJohannes Gutenberg-Universität Mainz
jgu.organisation.number7910
jgu.organisation.number2700
jgu.organisation.placeMainz
jgu.organisation.rorhttps://ror.org/023b0x485
jgu.pages.alternativeArt. 8876de
jgu.publisher.doi10.1038/s41598-017-09137-wde
jgu.publisher.issn2045-2322de
jgu.publisher.nameMacmillan Publishers Limited, part of Springer Naturede
jgu.publisher.placeLondonde
jgu.publisher.urihttp://dx.doi.org/10.1038/s41598-017-09137-wde
jgu.publisher.year2017
jgu.rights.accessrightsopenAccess
jgu.subject.ddccode610de
jgu.type.dinitypeArticleen_GB
jgu.type.resourceTextde
jgu.type.versionPublished versionde
opus.affiliatedEhlert, Tobias
opus.affiliatedTug, Suzan
opus.affiliatedBrahmer, Alexandra
opus.affiliatedHeid, Florian
opus.affiliatedWerner, Christian
opus.affiliatedKneist, Werner
opus.affiliatedLang, Hauke
opus.affiliatedSimon, Perikles
opus.date.modified2017-09-08T09:22:44Z
opus.identifier.opusid57256
opus.institute.number0436
opus.institute.number0418
opus.institute.number0208
opus.metadataonlyfalse
opus.organisation.stringFB 04: Medizin: Klinik und Poliklinik für Allgemein- und Abdominal-Chirurgiede_DE
opus.organisation.stringFB 04: Medizin: Klinik für Anästhesiologiede_DE
opus.organisation.stringFB 02: Sozialwissenschaften, Medien und Sport: Institut für Sportwissenschaftde_DE
opus.subject.dfgcode00-000
opus.type.contenttypeKeinede_DE
opus.type.contenttypeNoneen_EN

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