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Detection of KRAS, NRAS and BRAF mutations in liquid biopsy from patients with colorectal cancer

Ondraskova, K.; Cwik, M.; Horky, O.; Berkovcova, J.; Holcakova, J.; Bartosik, M.; Kazda, T.; Mrazova, K.; Uher, M.; Kiss, I.; Hrstka, R.

2025-10-09 oncology
10.1101/2025.10.08.25337549 medRxiv
Show abstract

ObjectivesCancer treatment relies heavily on accurate diagnosis and effective monitoring of the disease. These processes often involve invasive procedures, such as colonoscopy, to detect malignant tissues followed by molecular analyses to determine relevant biomarkers. This study aimed to evaluate the clinical performance of droplet digital PCR (ddPCR) for detecting KRAS, NRAS, and BRAF mutations in circulating tumor DNA (ctDNA) from colorectal cancer patients using liquid biopsy. MethodsctDNA was isolated from CRC patients (n = 110) and analyzed for KRAS, BRAF, and NRAS mutations. The ctDNA obtained through liquid biopsy was analyzed using ddPCR, and the findings were compared with sequencing data from tumor DNA archived in formalin fixed paraffin-embedded blocks (FFPE). ResultsFor KRAS mutations, ddPCR achieved a sensitivity of 72% and a specificity of 71.4%. However, when pooling all target mutations (KRAS, NRAS and BRAF), the overall sensitivity and specificity were lower, at 48.3% and 51.1%, respectively. ConclusionThe results of this study indicate that the ddPCR analysis of ctDNA may provide complementary information for the molecular diagnosis of CRC patients.

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