Prospective evaluation of two screening methods for molecular testing of metastatic melanoma: Diagnostic performance of BRAF V600E immunohistochemistry and of a NRAS-BRAF fully automated real-time PCR-based assay

• DOI: 10.1371/journal.pone.0221123
• Publication Date: 2019-08-15T17:44:31Z
• Authors: Audrey Vallée; Marie Denis-Musquer; Guillaume Herbreteau; Sandrine Théoleyre; Céline Bossard; Marc G. Denis
• Keywords: Concordance; Sanger sequencing; V600E; Mutation Testing; Molecular diagnostics

Screening for theranostic biomarkers is mandatory the therapeutic management of cutaneous melanoma. BRAF and NRAS genes must be tested in routine clinical practice. The methods used to identify these alterations sensitive detect mutant alleles a background wild type alleles, specific correct mutation. They should not require too much material, since some cases available samples are small biopsies. Finally, they also quick enough allow rapid patients. Sixty five consecutive formalin-fixed paraffin-embedded (FFPE) melanoma were prospectively mutations with VE1 (anti-BRAF V600E) antibody both Idylla NRAS-BRAF-EGFR S492R Mutation Assay cartridges. Results compared our laboratory practice, allele amplification and/or Sanger sequencing discordant confirmed by digital PCR. Excluding by-design-mutations, system failures DNA quantity or quality failures, IHC demonstrated an overall concordance 89% V600E mutation detection, gave 100% detection 92.1% when reference. When discrepancies observed, all results positive predictive value (PPV) was 82% negative (NPV) 92%. cartridges showed PPV NPV 87% respectively, detection. In conclusion, immunohistochemistry efficient detecting mutation, but further evaluated molecular approaches other mutations. Since 3 have been detected Assay, as substitute traditional conventional patient care process without expertise needed critical view produced results.