8. Rapid clinical BRAF V600 mutation testing using Idylla Platform

Rapid clinical testing is required as a standard of care to determine the treatment of patients with advanced cancers in particular for anaplastic thyroid carcinoma and melanoma. In this study, we evaluated the performances of the fast and fully automated Idylla BRAF Mutation Assay, performed on the Biocartis Idylla System, for the qualitative detection of BRAF V600E/E2/D and V600K/R/M mutations. In this study, 7 cell lines and 40 clinical specimens (20 FFPE sections, genomic DNA extracted from 8 FFPE, 1 FNA, 3 FNA washes, 6 peripheral blood and 2 bone marrow) with different diagnosis and known BRAF mutation status (20 positive, 20 negative) by NGS were evaluated. Idylla Explorer software was used for data analysis. Assay performance characteristics were established for clinical implementation. Limit of detection LOD (1%) was established based on serially diluted positive cell line and patient sample. An endogenous BRAF WT Cq of ≤35 was set as a threshold to check for adequacy of sample for amplification. Samples with endogenous WT Cq ≤32 demonstrated 1% LOD as compared to samples with WT Cq ≥32-≤35 showed 5% LOD. Any negative sample with Cq value higher than ≥35 will be repeated with more input material. Reproducibility studies performed using 3 positive and 3 negative samples showed 100% concordance. Idylla BRAF assay demonstrated 100% accuracy in tested positive and negative samples for BRAF mutation detection. In conclusion, Idylla BRAF assay offers rapid (< 2 hours) and reliable BRAF testing in routine and challenging specimens with limited material.