Key considerations for comprehensive validation of an RNA fusion NGS panel

• DOI: 10.1016/j.plabm.2020.e00173
• Publication Date: 2020-06-08T05:48:11Z
• Authors: Subit Barua; Gary Wang; Mahesh Mansukhani; Susan Hsiao; Helen Fernandes
• Keywords: 0301 basic medicine; Medicine (General); Precision medicine; Assay; Article; 3. Good health; Chemistry; 03 medical and health sciences; R5-920; 0302 clinical medicine; Next generation sequencing; RNA Fusions; Validation; Molecular oncology; QD1-999

Validation of RNA-based NGS assays for the detection therapeutically targetable gene fusions is challenging. Here, we report systematic validation and quality control monitoring our targeted fusion panel 17 clinically relevant transcripts across several tumor types. We implemented this RNA Fusion Panel as a reflex test tumors lacking DNA driver mutations.Forty-four formalin-fixed, paraffin-embedded (FFPE) or fresh-frozen lung, brain, soft tissue skin were used to determine accuracy assay. Additional fusion-positive specimens calibrated reference standard establish precision, reproducibility sensitivity All aspects validation, including metrics, performed according New York State guidelines.For panel, accuracy, precision studies above 99%. Reproducibility with helpful in identifying inconsistencies. The limit most was 50 copies. Application assay 450 samples mutations resulted 10% increase diagnostic yield minimal additional processing time.The validated provides clinical utility therapy selection patients solid tumors. By using sequential testing approach, complements hotspot variants many types time.