20.500.12556/DiRROS-25046 Somatic mutation detection in tumor tissue and matched cell-free DNA using PCR-based methods in pancreatic cancer patients undergoing upfront resection Somatic mutations in KRAS and TP53 are among the most common genetic alterations in pancreatic ductal adenocarcinoma (PDAC). Advances in PCR-based technologies now enable the detection of these mutations in tumor tissue and cell-free DNA (cfDNA), providing a minimally invasive approach to assess tumor burden. However, in resectable PDAC, circulating tumor DNA (ctDNA) may represent less than 0.1% of total cfDNA, requiring highly sensitive detection methods. The aim of our study was to assess two PCR-based assays—competitive allele-specific PCR (castPCR) and digital PCR (dPCR)—for detecting selected somatic mutations in tumor tissue, cfDNA, and extracellular vesicle-associated DNA (EV-DNA) from plasma. Matched primary tumor and preoperative plasma samples were collected from 50 patients undergoing upfront resection for PDAC. CastPCR was used for detecting selected KRAS, TP53, SMAD4, and CDKN2A mutations in tumor DNA. Additionally, dPCR was used to analyze KRAS and TP53 mutations in tumor DNA as well as cfDNA and EV-DNA. The concordance between both platforms was 71.4% for KRAS p.G12D and 58.3% for the analysis of TP53 p.R273H mutations in tumor tissue. However, dPCR detected these mutations in an additional 28.6% and 39.6% of samples, respectively. In cfDNA, dPCR identified KRAS p.G12D in 10.2% and TP53 p.R273H in 2.0% of samples. Mutation detection in EV-DNA was limited by low DNA yield. Both platforms proved effective for tumor DNA analysis, with dPCR offering greater sensitivity. Somatic mutation detection from liquid biopsy using dPCR further supports its potential utility in the preoperative setting. somatic mutation detection cell-free DNA liquid biopsy digital PCR pancreatic cancer okrivanje somatskih mutacij prosta celična DNK tekočinska biopsija digitalni PCR trebušna slinavka rak true false true Angleški jezik Slovenski jezik Neznano 2026-01-08 12:25:59 2026-01-08 12:25:59 2026-01-09 03:52:13 0000-00-00 00:00:00 2025 0 0 Nasl. z nasl. zaslona; Opis vira z dne 3. 12. 2025; str. 1-14 iss. 17 [article no.] 8518 Vol. 26 2025 0000-00-00 Zaloznikova Objavljeno NiDoloceno 0000-00-00 0000-00-00 0000-00-00 616-006 1422-0067 10.3390/ijms26178518 259680259 RAZ_Zavrtanik_Hana_2025.pdf RAZ_Zavrtanik_Hana_2025.pdf 1 CF886BA65B60E024604144634DC77135 4cb01d3bec39e0ac76e7e6d3c222e29284414ec400ef5613861705280c2ff37b eb9245a0-ec84-11f0-94a7-001a4af901a5 https://dirros.openscience.si/Dokument.php?lang=slv&id=37592 https://www.mdpi.com/1422-0067/26/17/8518 0 fbe5da6a-ec84-11f0-94a7-001a4af901a5 https://dirros.openscience.si/Dokument.php?lang=slv&id=37593 Univerzitetni klinični center Ljubljana 0 0 0