abstract = {Aim: Single biomarker diagnostic test of BRAFV600 locus in metastatic melanoma is mandatory for treatment decision; however, multiple-gene based techniques, such as targeted next-generation sequencing (NGS) are being used to maximize the number of patients that can benefit from a targeted therapy. The main objective of this study is to investigate whether an NGS panel could be adopted in routine clinical care for advanced melanoma. Methods: Patients diagnosed with advanced melanoma at our center from 2017 to 2019 were included. Presence of genetic alterations was performed using two methods: real-time polymerase chain reaction-based Idylla test (Biocartis) and NGS with the oncomine solid tumor DNA kit (Thermo Fisher Scientific). Total genomic DNA was extracted from formalin-fixed and paraffin embedded samples for sequencing. Results: A total of 155 samples were evaluated for molecular analysis but 40 samples (25.8{\%}) were inadequate for sequencing. The clinical utility of BRAFV600 real-time polymerase chain reaction and targeted-NGS was compared in 29 samples and a very good concordance was observed (Kappa = 0.89, 95{\%} confidence interval 0.68 ± 1.05). An oncogenic mutation by NGS was found in 75 samples (65{\%}) – 53{\%} of whom were candidates for personalized therapies. The most prevalent mutated genes were BRAF (39{\%}), TP53 (23{\%}), and NRAS (14{\%}). Other genes identified at lower incidence ({\textless} 5{\%}) were: PIK3CA, ERBB4, CTNNB1, STK11, FGFR1, SMAD4, KRAS, FGFR3, PTEN and AKT. Co-occurrence of oncogenic mutations was detected in 40{\%} of the samples. Among the mutations identified, TP53 was significantly more prevalent in men (men 31.8{\%} versus women 12.2{\%}, P = 0.03) and NRAS in women (men 9.1{\%} versus women 24.4{\%}, P = 0.03). Conclusions: Targeted-NGS testing is a feasible technique to implement in the routine clinical practice. Based on our results, NGS has provided more information on target-genes than RT-PCR technique, maximizing the benefit for patients with advanced melanoma.},
author = {Castillo, Paola and Marginet, Marta and Jares, Pedro and García, Mireia and Gonzalvo, Elena and Arance, Ana and García, Adriana and Alos, Llucia and Teixido, Cristina},
doi = {10.37349/etat.2020.00006},
issn = {2692-3114},
journal = {Exploration of Targeted Anti-tumor Therapy},
keywords = {BRAF,Idylla,melanoma,next-generation sequencing,real-time polymerase chain reaction},
number = {2},
pages = {101-108},
title = {{Implementation of an NGS panel for clinical practice in paraffin-embedded tissue samples from locally advanced and metastatic melanoma patients}},
url = {https://www.explorationpub.com/Journals/etat/Article/10026},
volume = {1},
year = {2020}
}