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Detection of BRAF mutations in malignant melanoma and colorectal cancer by SensiScreen(®) FFPE BRAF qPCR assay

Mutations in BRAF exon 15 lead to conformational changes in its activation loops, resulting in constitutively active BRAF proteins which are implicated in the development of several human cancer types. Different BRAF inhibitors have been developed and introduced in clinical practice. Identification...

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Autores principales: Sørensen, Anna Lahn, Guldmann-Christensen, Mariann, Børgesen, Michael, Petersen, Rasmus Koefoed, Flugt, Katharina, Duelund, Julie Mejer Holmgaard, Kyneb, Majbritt Hauge, Lorenzen, Jan, Pipó-Ollé, Emma, Epistolio, Samantha, Riva, Alice, Dazio, Giulia, Merlo, Elisabetta, Meyer, Tine, Christensen, Ulf Bech, Frattini, Milo
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9910728/
https://www.ncbi.nlm.nih.gov/pubmed/36758042
http://dx.doi.org/10.1371/journal.pone.0281558
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author Sørensen, Anna Lahn
Guldmann-Christensen, Mariann
Børgesen, Michael
Petersen, Rasmus Koefoed
Flugt, Katharina
Duelund, Julie Mejer Holmgaard
Kyneb, Majbritt Hauge
Lorenzen, Jan
Pipó-Ollé, Emma
Epistolio, Samantha
Riva, Alice
Dazio, Giulia
Merlo, Elisabetta
Meyer, Tine
Christensen, Ulf Bech
Frattini, Milo
author_facet Sørensen, Anna Lahn
Guldmann-Christensen, Mariann
Børgesen, Michael
Petersen, Rasmus Koefoed
Flugt, Katharina
Duelund, Julie Mejer Holmgaard
Kyneb, Majbritt Hauge
Lorenzen, Jan
Pipó-Ollé, Emma
Epistolio, Samantha
Riva, Alice
Dazio, Giulia
Merlo, Elisabetta
Meyer, Tine
Christensen, Ulf Bech
Frattini, Milo
author_sort Sørensen, Anna Lahn
collection PubMed
description Mutations in BRAF exon 15 lead to conformational changes in its activation loops, resulting in constitutively active BRAF proteins which are implicated in the development of several human cancer types. Different BRAF inhibitors have been developed and introduced in clinical practice. Identification of BRAF mutations influences the clinical evaluation, treatment, progression and for that reason a sensitive and specific identification of BRAF mutations is on request from the clinic. Here we present the SensiScreen(®) FFPE BRAF qPCR Assay that uses a novel real-time PCR-based method for BRAF mutation detection based on PentaBases proprietary DNA analogue technology designed to work on standard real-time PCR instruments. The SensiScreen(®) FFPE BRAF qPCR Assay displays high sensitivity, specificity, fast and easy-to-use. The SensiScreen(®) FFPE BRAF qPCR Assay was validated on two different FFPE tumour biopsy cohorts, one cohort included malignant melanoma patients previously analyzed by the Cobas® 4800 BRAF V600 Mutation Test, and one cohort from colorectal cancer patients previously analyzed by mutant-enriched PCR and direct sequencing. All BRAF mutant malignant melanoma patients were confirmed with the SensiScreen(®) FFPE BRAF qPCR Assay and additional four new mutations in the malignant melanoma cohort were identified. All the previously identified BRAF mutations in the colorectal cancer patients were confirmed, and additional three new mutations not identified with direct sequencing were detected. Also, one new BRAF mutation not previously identified with ME-PCR was found. Furthermore, the SensiScreen(®) FFPE BRAF qPCR Assay identified the specific change in the amino acid. The SensiScreen(®) FFPE BRAF qPCR Assay will contribute to a more specific, time and cost saving approach to better identify and characterize mutations in patients affected by cancer, and consequently permits a better BRAF characterization that is fundamental for therapy decision.
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spelling pubmed-99107282023-02-10 Detection of BRAF mutations in malignant melanoma and colorectal cancer by SensiScreen(®) FFPE BRAF qPCR assay Sørensen, Anna Lahn Guldmann-Christensen, Mariann Børgesen, Michael Petersen, Rasmus Koefoed Flugt, Katharina Duelund, Julie Mejer Holmgaard Kyneb, Majbritt Hauge Lorenzen, Jan Pipó-Ollé, Emma Epistolio, Samantha Riva, Alice Dazio, Giulia Merlo, Elisabetta Meyer, Tine Christensen, Ulf Bech Frattini, Milo PLoS One Research Article Mutations in BRAF exon 15 lead to conformational changes in its activation loops, resulting in constitutively active BRAF proteins which are implicated in the development of several human cancer types. Different BRAF inhibitors have been developed and introduced in clinical practice. Identification of BRAF mutations influences the clinical evaluation, treatment, progression and for that reason a sensitive and specific identification of BRAF mutations is on request from the clinic. Here we present the SensiScreen(®) FFPE BRAF qPCR Assay that uses a novel real-time PCR-based method for BRAF mutation detection based on PentaBases proprietary DNA analogue technology designed to work on standard real-time PCR instruments. The SensiScreen(®) FFPE BRAF qPCR Assay displays high sensitivity, specificity, fast and easy-to-use. The SensiScreen(®) FFPE BRAF qPCR Assay was validated on two different FFPE tumour biopsy cohorts, one cohort included malignant melanoma patients previously analyzed by the Cobas® 4800 BRAF V600 Mutation Test, and one cohort from colorectal cancer patients previously analyzed by mutant-enriched PCR and direct sequencing. All BRAF mutant malignant melanoma patients were confirmed with the SensiScreen(®) FFPE BRAF qPCR Assay and additional four new mutations in the malignant melanoma cohort were identified. All the previously identified BRAF mutations in the colorectal cancer patients were confirmed, and additional three new mutations not identified with direct sequencing were detected. Also, one new BRAF mutation not previously identified with ME-PCR was found. Furthermore, the SensiScreen(®) FFPE BRAF qPCR Assay identified the specific change in the amino acid. The SensiScreen(®) FFPE BRAF qPCR Assay will contribute to a more specific, time and cost saving approach to better identify and characterize mutations in patients affected by cancer, and consequently permits a better BRAF characterization that is fundamental for therapy decision. Public Library of Science 2023-02-09 /pmc/articles/PMC9910728/ /pubmed/36758042 http://dx.doi.org/10.1371/journal.pone.0281558 Text en © 2023 Sørensen et al https://creativecommons.org/licenses/by/4.0/This is an open access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle Research Article
Sørensen, Anna Lahn
Guldmann-Christensen, Mariann
Børgesen, Michael
Petersen, Rasmus Koefoed
Flugt, Katharina
Duelund, Julie Mejer Holmgaard
Kyneb, Majbritt Hauge
Lorenzen, Jan
Pipó-Ollé, Emma
Epistolio, Samantha
Riva, Alice
Dazio, Giulia
Merlo, Elisabetta
Meyer, Tine
Christensen, Ulf Bech
Frattini, Milo
Detection of BRAF mutations in malignant melanoma and colorectal cancer by SensiScreen(®) FFPE BRAF qPCR assay
title Detection of BRAF mutations in malignant melanoma and colorectal cancer by SensiScreen(®) FFPE BRAF qPCR assay
title_full Detection of BRAF mutations in malignant melanoma and colorectal cancer by SensiScreen(®) FFPE BRAF qPCR assay
title_fullStr Detection of BRAF mutations in malignant melanoma and colorectal cancer by SensiScreen(®) FFPE BRAF qPCR assay
title_full_unstemmed Detection of BRAF mutations in malignant melanoma and colorectal cancer by SensiScreen(®) FFPE BRAF qPCR assay
title_short Detection of BRAF mutations in malignant melanoma and colorectal cancer by SensiScreen(®) FFPE BRAF qPCR assay
title_sort detection of braf mutations in malignant melanoma and colorectal cancer by sensiscreen(®) ffpe braf qpcr assay
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9910728/
https://www.ncbi.nlm.nih.gov/pubmed/36758042
http://dx.doi.org/10.1371/journal.pone.0281558
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