2026-04-14T08:19:34Zhttps://recercat.cat/oai/requestoai:recercat.cat:11351/136982025-10-24T10:41:01Zcom_2072_378070com_2072_378040col_2072_378092
RECERCAT
author
Mayo de las Casas, Clara
author
Jantus Lewintre, Eloisa
author
Ortiz Reina, Sebastián
author
Drozdowskyj, Ana
author
Cerezuela-Fuentes, Pablo
author
MANZANO, JOSE LUIS
author
MUÑOZ COUSELO, EVA
2025-10-24T10:41:01Z
2025-10-24T10:41:01Z
2025-09-19T12:01:46Z2025-09-19T12:01:46Z2025
http://hdl.handle.net/11351/13698
Melanoma; Mutació BRAF; Biòpsia líquidaMelanoma; Mutación BRAF; Biopsia líquidaMelanoma; BRAF mutation; Liquid biopsyBackground
BRAF p.V600 mutation is the most frequent molecular driver alteration in melanoma. Detection of BRAF mutations in circulating-free DNA (cfDNA) reflects the shedding of tumor DNA and offers a potential non-invasive biomarker for disease monitoring and prognosis. However, the lack of standardized methodologies and inter-assay variability hinders its clinical implementation.
Methods
The sensitivity, agreement and concordance of seven BRAF mutation detection assays were assessed across four laboratories. BRAF p.V600 mutation in pretreatment plasma samples was analyzed in 51 patients diagnosed with advanced stage melanoma using two digital PCR-based assays (droplet digital PCR -ddPCR- Bio-Rad and microfluidic digital PCR -Absolute Q, ThermoFisher Scientific-), three RT-PCR based assays (Idylla®, Cobas®, PNA-Q-PCR) and two NGS based assays (Oncomine™ Pan-Cancer Cell-Free Assay and Illumina Platforms).
Results
digital PCR-based assays and Cobas® exhibited the highest sensitivity (51.0 %), followed by NGS Illumina® (45.1 %), Oncomine NGS / PNA-Q-PCR (43.1 %) and Idylla® (37.2 %). Results of different techniques showed a moderate to strong agreement, except for the comparison of Cobas with Idylla that was poor (Kappa=0.57). There was near-perfect agreement on detection of BRAF mutation between both NGS platforms and the NGS Illumina® with PNA-Q-PCR (Kappa = 0.92). Concordance of the quantitative results in terms of mutant allele frequency was near-perfect between NGS Illumina and ddPCR Bio-Rad assays (ICC = 0.99).
Conclusions
Our study demonstrates substantial agreement among multiple cfDNA BRAF mutation detection assays, particularly between NGS and digital PCR assays. These findings support the potential utility of different techniques for BRAF testing in cfDNA.This study was supported by the Novartis Group.
Attribution 4.0 International http://creativecommons.org/licenses/by/4.0/ info:eu-repo/semantics/openAccess
Anomalies cromosòmiques
Melanoma - Aspectes genètics
Melanoma - Prognosi
PHENOMENA AND PROCESSES::Genetic Phenomena::Genetic Variation::Mutation
DISEASES::Neoplasms::Neoplasms by Histologic Type::Neoplasms, Germ Cell and Embryonal::Neuroectodermal Tumors::Neuroendocrine Tumors::Melanoma
Other subheadings::Other subheadings::Other subheadings::/genetics
CHEMICALS AND DRUGS::Nucleic Acids, Nucleotides, and Nucleosides::Nucleic Acids::Cell-Free Nucleic Acids
CHEMICALS AND DRUGS::Enzymes and Coenzymes::Enzymes::Transferases::Phosphotransferases::Phosphotransferases (Alcohol Group Acceptor)::Protein Kinases::Protein-Serine-Threonine Kinases::MAP Kinase Kinase Kinases::raf Kinases::Proto-Oncogene Proteins c-raf
ANALYTICAL, DIAGNOSTIC AND THERAPEUTIC TECHNIQUES, AND EQUIPMENT::Diagnosis::Prognosis
FENÓMENOS Y PROCESOS::fenómenos genéticos::variación genética::mutación
ENFERMEDADES::neoplasias::neoplasias por tipo histológico::neoplasias de células germinales y embrionarias::tumores neuroectodérmicos::tumores neuroendocrinos::melanoma
Otros calificadores::Otros calificadores::Otros calificadores::/genética
COMPUESTOS QUÍMICOS Y DROGAS::nucleótidos y nucleósidos de ácidos nucleicos::ácidos nucleicos::ácidos nucleicos libres de células
COMPUESTOS QUÍMICOS Y DROGAS::enzimas y coenzimas::enzimas::transferasas::fosfotransferasas::fosfotransferasas (grupo alcohol aceptor)::proteína cinasas::proteína-serina-treonina cinasas::MAP cinasa cinasa cinasas::cinasas raf::proteínas protooncogénicas c-raf
TÉCNICAS Y EQUIPOS ANALÍTICOS, DIAGNÓSTICOS Y TERAPÉUTICOS::diagnóstico::pronóstico
Cross-method comparison for BRAF p.V600 mutation cfDNA testing in Melanoma: BRAFI study
info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion