The study introduces a structured methodology for the identification of molecular targets that accurately classify medulloblastoma subgroups: WNT, SHH, Group 3 (G3) and Group 4 (G4). An artificial neural network (ANN) model trained on microarray gene expression data determined minimal gene combinations for each subgroup. The classification achieved an average accuracy of 96%, demonstrating the effectiveness of the proposed approach. Feature selection using the Kruskal-Wallis and tests revealed statistically relevant genes contributing to subgroup discrimination. Reverse transcription followed by digital Polymerase Chain Reaction (dPCR) measured the expression levels of a subset of these genes in tumor samples, validating the computational predictions with experimental evidence. The integration of machine learning and molecular quantification provides a reproducible framework for medulloblastoma subgroup classification supported by both statistical and experimental consistency.