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ARTICLE

Detection of KRAS, NRAS and BRAF Mutations in Liquid Biopsy from Patients with Colorectal Cancer

Katerina Ondraskova1,2, Matous Cwik3, Ondrej Horky4, Jitka Berkovcova4, Jitka Holcakova1, Martin Bartosik1, Tomas Kazda5, Klara Mrazova1,6, Michal Uher7, Igor Kiss3, Roman Hrstka1,3,*
1 Research Centre for Applied Molecular Oncology (RECAMO), Masaryk Memorial Cancer Institute, Zluty kopec 7, Brno, 656 53, Czech Republic
2 National Centre for Biomolecular Research Faculty of Science, Masaryk University, Kamenice 5, Brno, 625 00, Czech Republic
3 Department of Comprehensive Cancer Care and Faculty of Medicine, Masaryk Memorial Cancer Institute and Masaryk University, Zluty kopec 7, Brno, 656 53, Czech Republic
4 Department of Pathology, Masaryk Memorial Cancer Institute, Zluty kopec 7, Brno, 656 53, Czech Republic
5 Department of Radiation Oncology, Masaryk Memorial Cancer Institute, Zluty kopec 7, Brno, 656 53, Czech Republic
6 Department of Biochemistry, Faculty of Science, Masaryk University, Kamenice 5, Brno, 625 00, Czech Republic
7 Department of Health Information, Masaryk Memorial Cancer Institute, Zluty kopec 7, Brno, 656 53, Czech Republic
* Corresponding Author: Roman Hrstka. Email: email

Oncology Research https://doi.org/10.32604/or.2025.070116

Received 08 July 2025; Accepted 11 December 2025; Published online 29 December 2025

Abstract

Objectives: Cancer 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 Kirsten Rat Sarcoma Viral Proto-Oncogene (KRAS), Neuroblastoma RAS Viral Oncogene Homolog (NRAS), and B-Raf Murine Sarcoma Viral Oncogene Homolog B (BRAF) mutations in circulating tumor DNA (ctDNA) from colorectal cancer patients using liquid biopsy. Methods: ctDNA was isolated from colorectal cancer (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 (FFPE) blocks. Results: For KRAS mutations, ddPCR achieved a sensitivity of 72.0% 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. Conclusion: The results of this study indicate that the ddPCR analysis of ctDNA may provide complementary information for the molecular diagnosis of CRC patients.

Keywords

Liquid biopsy; colorectal cancer (CRC); droplet digital PCR (ddPCR); Kirsten Rat Sarcoma Viral Proto-Oncogene (KRAS) mutation

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