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Comparison of BRAF Mutation Screening Strategies in a Large Real-Life Series of Advanced Melanoma Patients

Malignant melanoma (MM) is one of the deadliest skin cancers. BRAF mutation status plays a predominant role in the management of MM patients. The aim of this study was to compare BRAF mutational testing performed by conventional nucleotide sequencing approaches with either real-time polymerase chain...

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Autores principales: Colombino, Maria, Rozzo, Carla, Paliogiannis, Panagiotis, Casula, Milena, Manca, Antonella, Doneddu, Valentina, Fedeli, Maria Antonietta, Sini, Maria Cristina, Palomba, Grazia, Pisano, Marina, Ascierto, Paolo A., Caracò, Corrado, Lissia, Amelia, Cossu, Antonio, Palmieri, Giuseppe
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7464760/
https://www.ncbi.nlm.nih.gov/pubmed/32751423
http://dx.doi.org/10.3390/jcm9082430
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author Colombino, Maria
Rozzo, Carla
Paliogiannis, Panagiotis
Casula, Milena
Manca, Antonella
Doneddu, Valentina
Fedeli, Maria Antonietta
Sini, Maria Cristina
Palomba, Grazia
Pisano, Marina
Ascierto, Paolo A.
Caracò, Corrado
Lissia, Amelia
Cossu, Antonio
Palmieri, Giuseppe
author_facet Colombino, Maria
Rozzo, Carla
Paliogiannis, Panagiotis
Casula, Milena
Manca, Antonella
Doneddu, Valentina
Fedeli, Maria Antonietta
Sini, Maria Cristina
Palomba, Grazia
Pisano, Marina
Ascierto, Paolo A.
Caracò, Corrado
Lissia, Amelia
Cossu, Antonio
Palmieri, Giuseppe
author_sort Colombino, Maria
collection PubMed
description Malignant melanoma (MM) is one of the deadliest skin cancers. BRAF mutation status plays a predominant role in the management of MM patients. The aim of this study was to compare BRAF mutational testing performed by conventional nucleotide sequencing approaches with either real-time polymerase chain reaction (rtPCR) or next-generation sequencing (NGS) assays in a real-life, hospital-based series of advanced MM patients. Consecutive patients with AJCC (American Joint Committee on Cancer) stage IIIC and IV MM from Sardinia, Italy, who were referred for molecular testing, were enrolled into the study. Initial screening was performed to assess the mutational status of the BRAF and NRAS genes, using the conventional methodologies recognized by the nationwide guidelines, at the time of the molecular classification, required by clinicians: at the beginning, Sanger-based sequencing (SS) and, after, pyrosequencing. The present study was then focused on BRAF mutation detecting approaches only. BRAF wild-type cases with available tissue and adequate DNA were further tested with rtPCR (Idylla™) and NGS assays. Globally, 319 patients were included in the study; pathogenic BRAF mutations were found in 144 (45.1%) cases examined with initial screening. The rtPCR detected 11 (16.2%) and 3 (4.8%) additional BRAF mutations after SS and pyrosequencing, respectively. NGS detected one additional BRAF-mutated case (2.1%) among 48 wild-type cases previously tested with pyrosequencing and rtPCR. Our study evidenced that rtPCR and NGS were able to detect additional BRAF mutant cases in comparison with conventional sequencing methods; therefore, we argue for the preferential utilization of the aforementioned assays (NGS and rtPCR) in clinical practice, to eradicate false-negative cases and improve the accuracy of BRAF detection.
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spelling pubmed-74647602020-09-04 Comparison of BRAF Mutation Screening Strategies in a Large Real-Life Series of Advanced Melanoma Patients Colombino, Maria Rozzo, Carla Paliogiannis, Panagiotis Casula, Milena Manca, Antonella Doneddu, Valentina Fedeli, Maria Antonietta Sini, Maria Cristina Palomba, Grazia Pisano, Marina Ascierto, Paolo A. Caracò, Corrado Lissia, Amelia Cossu, Antonio Palmieri, Giuseppe J Clin Med Article Malignant melanoma (MM) is one of the deadliest skin cancers. BRAF mutation status plays a predominant role in the management of MM patients. The aim of this study was to compare BRAF mutational testing performed by conventional nucleotide sequencing approaches with either real-time polymerase chain reaction (rtPCR) or next-generation sequencing (NGS) assays in a real-life, hospital-based series of advanced MM patients. Consecutive patients with AJCC (American Joint Committee on Cancer) stage IIIC and IV MM from Sardinia, Italy, who were referred for molecular testing, were enrolled into the study. Initial screening was performed to assess the mutational status of the BRAF and NRAS genes, using the conventional methodologies recognized by the nationwide guidelines, at the time of the molecular classification, required by clinicians: at the beginning, Sanger-based sequencing (SS) and, after, pyrosequencing. The present study was then focused on BRAF mutation detecting approaches only. BRAF wild-type cases with available tissue and adequate DNA were further tested with rtPCR (Idylla™) and NGS assays. Globally, 319 patients were included in the study; pathogenic BRAF mutations were found in 144 (45.1%) cases examined with initial screening. The rtPCR detected 11 (16.2%) and 3 (4.8%) additional BRAF mutations after SS and pyrosequencing, respectively. NGS detected one additional BRAF-mutated case (2.1%) among 48 wild-type cases previously tested with pyrosequencing and rtPCR. Our study evidenced that rtPCR and NGS were able to detect additional BRAF mutant cases in comparison with conventional sequencing methods; therefore, we argue for the preferential utilization of the aforementioned assays (NGS and rtPCR) in clinical practice, to eradicate false-negative cases and improve the accuracy of BRAF detection. MDPI 2020-07-30 /pmc/articles/PMC7464760/ /pubmed/32751423 http://dx.doi.org/10.3390/jcm9082430 Text en © 2020 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Colombino, Maria
Rozzo, Carla
Paliogiannis, Panagiotis
Casula, Milena
Manca, Antonella
Doneddu, Valentina
Fedeli, Maria Antonietta
Sini, Maria Cristina
Palomba, Grazia
Pisano, Marina
Ascierto, Paolo A.
Caracò, Corrado
Lissia, Amelia
Cossu, Antonio
Palmieri, Giuseppe
Comparison of BRAF Mutation Screening Strategies in a Large Real-Life Series of Advanced Melanoma Patients
title Comparison of BRAF Mutation Screening Strategies in a Large Real-Life Series of Advanced Melanoma Patients
title_full Comparison of BRAF Mutation Screening Strategies in a Large Real-Life Series of Advanced Melanoma Patients
title_fullStr Comparison of BRAF Mutation Screening Strategies in a Large Real-Life Series of Advanced Melanoma Patients
title_full_unstemmed Comparison of BRAF Mutation Screening Strategies in a Large Real-Life Series of Advanced Melanoma Patients
title_short Comparison of BRAF Mutation Screening Strategies in a Large Real-Life Series of Advanced Melanoma Patients
title_sort comparison of braf mutation screening strategies in a large real-life series of advanced melanoma patients
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7464760/
https://www.ncbi.nlm.nih.gov/pubmed/32751423
http://dx.doi.org/10.3390/jcm9082430
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