Detection of the BRAF V600E Mutation in Liquid Biopsy Using the CRISPR/Cas12a System

The goal of this prospective, low-interventional study is to develop and optimize a highly sensitive and specific method for the detection of the BRAF V600E mutation in liquid biopsy samples from patients with unresectable metastatic melanoma after disease relapse. The method will be based on a CRISPR/Cas12a system combined with isothermal amplification and endpoint electrochemical detection using electrode chips and low-cost instrumentation.

Inclusion Criteria: * Age ≥ 18 years. * Subject willing and able to provide written informed consent for biological sample collection. * Ability to communicate effectively with the investigator in the local language, as well as the ability to understand and comply with study requirements. * Histologically confirmed malignant melanoma. * Radiologically or histologically confirmed disease relapse. * Feasibility/possibility of peripheral blood sampling for ctDNA analysis. * Documented, known \*BRAF\* mutation status (wild-type vs. V600E). Exclusion Criteria: * Operable disease relapse. * Age under 18 years. * Pregnant or breastfeeding women. * Patients receiving palliative care with an estimated life expectancy ≤ 3 months. * Severe alcohol and/or drug abuse during therapy. * Inability to obtain a valid biological sample for molecular analysis