Circulating cell-free DNA (cfDNA) and its tumour-derived circulating tumour DNA (ctDNA) fraction are considered an innovative prognostic and predictive biomarker in oncological diagnostics. Many studies have demonstrated higher levels of cfDNA concentration and integrity, as an indicator of the amount of ctDNA in cfDNA, in body fluids from patients with cancer diseases in comparison with healthy individuals, which suggest its potential as an effective biomarker for monitoring of the tumour dynamics. This study focused on optimisation and validation of measurement methods later used for analysis of cfDNA concentration and integrity in blood samples from patients with four different solid cancers. Two different commercial isolation kits have been tested in plasma and serum samples. Quantitative real-time polymerase reaction (qPCR) and PicoGreen dsDNA assay were optimized to effectively quantify low concentrations of cfDNA, subsequently compared to each other and to droplet digital PCR assay tested on selected samples. The concentration and integrity of cfDNA from plasma samples of breast, ovarian, colorectal and pancreatic cancer patients were evaluated. Higher amounts of cfDNA were obtained by the QIAamp Circulating Nucleic Acid isolation kit (Qiagen) in comparison to Plasma/Serum Cell-Free...
| Identifer | oai:union.ndltd.org:nusl.cz/oai:invenio.nusl.cz:405209 |
| Date | January 2019 |
| Creators | Buranovská, Katarína |
| Contributors | Souček, Pavel, Němcová, Vlasta |
| Source Sets | Czech ETDs |
| Language | English |
| Detected Language | English |
| Type | info:eu-repo/semantics/masterThesis |
| Rights | info:eu-repo/semantics/restrictedAccess |
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