Establishing PNB-qPCR for quantifying minimal ctDNA concentrations during tumour resection
- Publikationstyp:
- Zeitschriftenaufsatz
- Metadaten:
-
- Autoren
- Tobias Ehlert
- Suzan Tug
- Alexandra Brahmer
- Vanessa Neef
- Florian Heid
- Christian Werner
- Boris Jansen-Winkeln
- Werner Kneist
- Hauke Lang
- Ines Gockel
- Perikles Simon
- Sammlungen
- metadata
- ISSN
- 2045-2322
- Zeitschrift
- Scientific reports
- Schlüsselwörter
- 610 Medizin
- 610 Medical sciences
- Sprache
- eng
- Paginierung
- Art. 8876
- Datum der Veröffentlichung
- 2017
- Herausgeber
- Macmillan Publishers Limited, part of Springer Nature
- Herausgeber URL
- http://dx.doi.org/10.1038/s41598-017-09137-w
- Datum der Datenerfassung
- 2020
- Datum, an dem der Datensatz öffentlich gemacht wurde
- 2020
- Zugang
- Public
- Titel
- Establishing PNB-qPCR for quantifying minimal ctDNA concentrations during tumour resection
- Ausgabe der Zeitschrift
- 7
Datenquelle: METADATA.UB
- Andere Metadatenquellen:
-
- Autoren
- T Ehlert
- S Tug
- A Brahmer
- V Neef
- F Heid
- C Werner
- B Jansen-Winkeln
- W Kneist
- H Lang
- I Gockel
- P Simon
- Autoren-URL
- https://www.webofscience.com/api/gateway?GWVersion=2&SrcApp=fis-test-1&SrcAuth=WosAPI&KeyUT=WOS:000408107000010&DestLinkType=FullRecord&DestApp=WOS_CPL
- DOI
- 10.1038/s41598-017-09137-w
- Externe Identifier
- Clarivate Analytics Document Solution ID: FE3HM
- PubMed Identifier: 28827745
- ISSN
- 2045-2322
- Zeitschrift
- SCIENTIFIC REPORTS
- Artikelnummer
- ARTN 8876
- Datum der Veröffentlichung
- 2017
- Status
- Published
- Titel
- Establishing PNB-qPCR for quantifying minimal ctDNA concentrations during tumour resection
- Sub types
- Article
- Ausgabe der Zeitschrift
- 7
Datenquelle: Web of Science (Lite)
- Abstract
- <jats:title>Abstract</jats:title><jats:p>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 <jats:italic>KRAS</jats:italic> exon 2. Using this approach, we were able to characterize ctDNA and total cell-free DNA (cfDNA) kinetics by selective amplification of <jats:italic>KRAS</jats:italic> 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.</jats:p>
- Autoren
- T Ehlert
- S Tug
- A Brahmer
- V Neef
- F Heid
- C Werner
- B Jansen-Winkeln
- W Kneist
- H Lang
- I Gockel
- P Simon
- DOI
- 10.1038/s41598-017-09137-w
- eISSN
- 2045-2322
- Ausgabe der Veröffentlichung
- 1
- Zeitschrift
- Scientific Reports
- Sprache
- en
- Artikelnummer
- 8876
- Online publication date
- 2017
- Status
- Published online
- Herausgeber
- Springer Science and Business Media LLC
- Herausgeber URL
- http://dx.doi.org/10.1038/s41598-017-09137-w
- Datum der Datenerfassung
- 2022
- Titel
- Establishing PNB-qPCR for quantifying minimal ctDNA concentrations during tumour resection
- Ausgabe der Zeitschrift
- 7
Datenquelle: Crossref
- 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.
- Addresses
- Department of Sports Medicine, Rehabilitation and Disease Prevention, Faculty of Social Science, Media and Sport, Johannes Gutenberg-University Mainz, Mainz, Germany.
- Autoren
- T Ehlert
- S Tug
- A Brahmer
- V Neef
- F Heid
- C Werner
- B Jansen-Winkeln
- W Kneist
- H Lang
- I Gockel
- P Simon
- DOI
- 10.1038/s41598-017-09137-w
- eISSN
- 2045-2322
- Externe Identifier
- PubMed Identifier: 28827745
- PubMed Central ID: PMC5566323
- Open access
- true
- ISSN
- 2045-2322
- Ausgabe der Veröffentlichung
- 1
- Zeitschrift
- Scientific reports
- Schlüsselwörter
- Humans
- Neoplasms
- DNA, Neoplasm
- Sensitivity and Specificity
- DNA Mutational Analysis
- Mutation
- Proto-Oncogene Proteins p21(ras)
- Real-Time Polymerase Chain Reaction
- Biomarkers, Tumor
- Circulating Tumor DNA
- Sprache
- eng
- Medium
- Electronic
- Online publication date
- 2017
- Open access status
- Open Access
- Paginierung
- 8876
- Datum der Veröffentlichung
- 2017
- Status
- Published
- Publisher licence
- CC BY
- Datum der Datenerfassung
- 2017
- Titel
- Establishing PNB-qPCR for quantifying minimal ctDNA concentrations during tumour resection.
- Sub types
- research-article
- Journal Article
- Ausgabe der Zeitschrift
- 7
Files
https://www.nature.com/articles/s41598-017-09137-w.pdf https://europepmc.org/articles/PMC5566323?pdf=render
Datenquelle: Europe PubMed Central
- 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.
- Date of acceptance
- 2017
- Autoren
- T Ehlert
- S Tug
- A Brahmer
- V Neef
- F Heid
- C Werner
- B Jansen-Winkeln
- W Kneist
- H Lang
- I Gockel
- P Simon
- Autoren-URL
- https://www.ncbi.nlm.nih.gov/pubmed/28827745
- DOI
- 10.1038/s41598-017-09137-w
- eISSN
- 2045-2322
- Externe Identifier
- PubMed Central ID: PMC5566323
- Ausgabe der Veröffentlichung
- 1
- Zeitschrift
- Sci Rep
- Schlüsselwörter
- Biomarkers, Tumor
- Circulating Tumor DNA
- DNA Mutational Analysis
- DNA, Neoplasm
- Humans
- Mutation
- Neoplasms
- Proto-Oncogene Proteins p21(ras)
- Real-Time Polymerase Chain Reaction
- Sensitivity and Specificity
- Sprache
- eng
- Country
- England
- Paginierung
- 8876
- PII
- 10.1038/s41598-017-09137-w
- Datum der Veröffentlichung
- 2017
- Status
- Published online
- Datum, an dem der Datensatz öffentlich gemacht wurde
- 2019
- Titel
- Establishing PNB-qPCR for quantifying minimal ctDNA concentrations during tumour resection.
- Sub types
- Journal Article
- Ausgabe der Zeitschrift
- 7
Datenquelle: PubMed
- Author's licence
- CC-BY
- Autoren
- Tobias Ehlert
- Suzan Tug
- Alexandra Brahmer
- Vanessa Neef
- Florian Heid
- Christian Werner
- Boris Jansen-Winkeln
- Werner Kneist
- Hauke Lang
- Ines Gockel
- Perikles Simon
- Hosting institution
- Universitätsbibliothek Mainz
- Sammlungen
- DFG-OA-Publizieren (2012 - 2017)
- Resource version
- Published version
- DOI
- 10.1038/s41598-017-09137-w
- Funding acknowledgements
- DFG, Open Access-Publizieren Universität Mainz / Universitätsmedizin
- File(s) embargoed
- false
- Open access
- true
- ISSN
- 2045-2322
- Zeitschrift
- Scientific reports
- Schlüsselwörter
- 610 Medizin
- 610 Medical sciences
- Sprache
- eng
- Open access status
- Open Access
- Paginierung
- Art. 8876
- Datum der Veröffentlichung
- 2017
- Public URL
- https://openscience.ub.uni-mainz.de/handle/20.500.12030/7960
- Herausgeber
- Macmillan Publishers Limited, part of Springer Nature
- Herausgeber URL
- http://dx.doi.org/10.1038/s41598-017-09137-w
- Datum der Datenerfassung
- 2022
- Datum, an dem der Datensatz öffentlich gemacht wurde
- 2022
- Zugang
- Public
- Titel
- Establishing PNB-qPCR for quantifying minimal ctDNA concentrations during tumour resection
- Ausgabe der Zeitschrift
- 7
Files
establishing_pnbqpcr_for_quan-20220925142339407.pdf
Datenquelle: OPENSCIENCE.UB
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