Multi-center evaluation of the novel fully-automated PCR-based Idylla™ BRAF Mutation Test on formalin-fixed paraffin-embedded tissue of malignant melanoma

Abstract

The advent of BRAF-targeted therapies led to increased survival in patients with metastatic melanomas harboring a BRAF V600 mutation (implicated in 46-48% of malignant melanomas). The Idylla(™) System (Idylla(™)), i.e., the real-time-PCR-based Idylla(™) BRAF Mutation Test performed on the fully-automated Idylla(™) platform, enables detection of the most frequent BRAF V600 mutations (V600E/E2/D, V600K/R/M) in tumor material within approximately 90 min and with 1% detection limit. Idylla(™) performance was determined in a multi-center study by analyzing BRAF mutational status of 148 archival formalin-fixed paraffin-embedded (FFPE) tumor samples from malignant melanoma patients, and comparing Idylla(™) results with assessments made by commercial or in-house routine diagnostic methods. Of the 148 samples analyzed, Idylla(™) initially recorded 7 insufficient DNA input calls and 15 results discordant with routine method results. Further analysis learned that the quality of 8 samples was insufficient for Idylla(™) testing, 1 sample had an invalid routine test result, and Idylla(™) results were confirmed in 10 samples. Hence, Idylla(™) identified all mutations present, including 7 not identified by routine methods. Idylla(™) enables fully automated BRAF V600 testing directly on FFPE tumor tissue with increased sensitivity, ease-of-use, and much shorter turnaround time compared to existing diagnostic tests, making it a tool for rapid, simple and highly reliable analysis of therapeutically relevant BRAF mutations, in particular for diagnostic units without molecular expertise and infrastructure.

Fully automated PCR detection of KRAS mutations on pancreatic endoscopic ultrasound fine-needle aspirates

“…However, KRAS testing is difficult to be implemented in cytopathology laboratories, with little expertise in molecular biology procedures and in-house validation of a laboratory-developed methods. The Idylla KRAS Mutation Test, a rapid and fully automated CE-IVD (European Conformity-In Vitro Diagnostic)-certified test, can represent a valid option for a wider number of pathological centres 16 17 20. The assay has been designed by the manufacturer to process FFPE tissue sections using high-frequency intensity-focused ultrasound technology to obtain amplifiable DNA 16 17 20.…”

“…This test, based on allele-specific RT-PCR and performed on the fully automated Idylla platform, provides sample to result functionality, enabling method standardisation even in those diagnostic units without molecular expertise and infrastructure 16 17. The aim of this study was to validate this novel technology on indeterminate pancreatic EUS-FNA.…”

The usefulness of a novel fully automated PCR-based Idylla test for detection of the BRAF V600E mutation in thyroid tissue: comparison with PNA-clamping PCR, real-time PCR and pyrosequencing

“…The Idylla test has a benefit to perform sample processing with a minimum 3 mm 2 area with 20 μm tissue sections and yields results with the least amount of FFPE thyroid samples among the applied methods. Thyroid FFPE samples with less than minimum and more than maximum tissue area as specified in the Idylla instruction could generate BRAF V600E mutation, unlike the previous results on malignant melanoma samples 20. The validated tissue amount for the Idylla BRAF test might be different depending on the tissue types and the presence of pigment.…”

Superior Multiplexing Capacity of PlexPrimers Enables Sensitive and Specific Detection of SNPs and Clustered Mutations in qPCR

“…Their study concluded the Idylla assay and platform enabled very rapid decision making with high sensitivity, large reference range and ease of use even in less experienced laboratories [19]. Likewise, Solassol et al reported similar clinical utility and convenience reporting that the Idylla KRAS Mutation Test afforded simple, highly reliable and rapid routine determination of KRAS mutational status to guide colorectal therapy [20]. …”