{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,13]],"date-time":"2026-06-13T11:01:52Z","timestamp":1781348512840,"version":"3.54.1"},"reference-count":28,"publisher":"Public Library of Science (PLoS)","issue":"2","license":[{"start":{"date-parts":[[2023,2,9]],"date-time":"2023-02-09T00:00:00Z","timestamp":1675900800000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/100013297","name":"Eurostars","doi-asserted-by":"publisher","award":["E10440"],"award-info":[{"award-number":["E10440"]}],"id":[{"id":"10.13039\/100013297","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["www.plosone.org"],"crossmark-restriction":false},"short-container-title":["PLoS ONE"],"abstract":"<jats:p>Mutations in <jats:italic>BRAF<\/jats:italic> 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 <jats:italic>BRAF<\/jats:italic> mutations influences the clinical evaluation, treatment, progression and for that reason a sensitive and specific identification of <jats:italic>BRAF<\/jats:italic> mutations is on request from the clinic. Here we present the SensiScreen<jats:sup>\u00ae<\/jats:sup> FFPE BRAF qPCR Assay that uses a novel real-time PCR-based method for <jats:italic>BRAF<\/jats:italic> mutation detection based on PentaBases proprietary DNA analogue technology designed to work on standard real-time PCR instruments. The SensiScreen<jats:sup>\u00ae<\/jats:sup> FFPE BRAF qPCR Assay displays high sensitivity, specificity, fast and easy-to-use. The SensiScreen<jats:sup>\u00ae<\/jats:sup> FFPE BRAF qPCR Assay was validated on two different FFPE tumour biopsy cohorts, one cohort included malignant melanoma patients previously analyzed by the Cobas\u00ae 4800 <jats:italic>BRAF<\/jats:italic> V600 Mutation Test, and one cohort from colorectal cancer patients previously analyzed by mutant-enriched PCR and direct sequencing. All <jats:italic>BRAF<\/jats:italic> mutant malignant melanoma patients were confirmed with the SensiScreen<jats:sup>\u00ae<\/jats:sup> FFPE BRAF qPCR Assay and additional four new mutations in the malignant melanoma cohort were identified. All the previously identified <jats:italic>BRAF<\/jats:italic> mutations in the colorectal cancer patients were confirmed, and additional three new mutations not identified with direct sequencing were detected. Also, one new <jats:italic>BRAF<\/jats:italic> mutation not previously identified with ME-PCR was found. Furthermore, the SensiScreen<jats:sup>\u00ae<\/jats:sup> FFPE BRAF qPCR Assay identified the specific change in the amino acid. The SensiScreen<jats:sup>\u00ae<\/jats:sup> 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 <jats:italic>BRAF<\/jats:italic> characterization that is fundamental for therapy decision.<\/jats:p>","DOI":"10.1371\/journal.pone.0281558","type":"journal-article","created":{"date-parts":[[2023,2,9]],"date-time":"2023-02-09T18:50:31Z","timestamp":1675968631000},"page":"e0281558","update-policy":"https:\/\/doi.org\/10.1371\/journal.pone.corrections_policy","source":"Crossref","is-referenced-by-count":1,"title":["Detection of BRAF mutations in malignant melanoma and colorectal cancer by SensiScreen\u00ae FFPE BRAF qPCR assay"],"prefix":"10.1371","volume":"18","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-9002-8995","authenticated-orcid":true,"given":"Anna Lahn","family":"S\u00f8rensen","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mariann","family":"Guldmann-Christensen","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Michael","family":"B\u00f8rgesen","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rasmus Koefoed","family":"Petersen","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Katharina","family":"Flugt","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Julie Mejer Holmgaard","family":"Duelund","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Majbritt Hauge","family":"Kyneb","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jan","family":"Lorenzen","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Emma","family":"Pip\u00f3-Oll\u00e9","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1236-2722","authenticated-orcid":true,"given":"Samantha","family":"Epistolio","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alice","family":"Riva","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Giulia","family":"Dazio","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Elisabetta","family":"Merlo","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tine","family":"Meyer","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ulf Bech","family":"Christensen","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Milo","family":"Frattini","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"340","published-online":{"date-parts":[[2023,2,9]]},"reference":[{"key":"pone.0281558.ref001","author":"H Davies","year":"2002","journal-title":"Mutations of the BRAF gene in human cancer"},{"key":"pone.0281558.ref002","first-page":"329","article-title":"Targeting alterations in the RAF\u2013MEK pathway","volume-title":"Cancer Discovery.","author":"R Yaeger","year":"2019"},{"key":"pone.0281558.ref003","first-page":"12","article-title":"Contemporary treatment approaches for metastatic colorectal cancer driven by BRAF V600 mutations","author":"O Kanaty","year":"2020","journal-title":"rld J Gastrointest Oncol"},{"key":"pone.0281558.ref004","first-page":"48","article-title":"Dabrafenib in advanced melanoma with BRAF V600E mutation.","author":"J Abraham","year":"2014","journal-title":"Journal of Community and Supportive Oncology. 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Available: www.jco.org"},{"key":"pone.0281558.ref007","doi-asserted-by":"crossref","first-page":"2035","DOI":"10.1158\/1078-0432.CCR-13-2054","article-title":"Dabrafenib and Trametinib, alone and in combination for BRAF-Mutant metastatic melanoma","volume":"20","author":"AM Menzies","year":"2014","journal-title":"Clinical Cancer Research"},{"key":"pone.0281558.ref008","article-title":"Current and Future Roles of Targeted Therapy and Immunotherapy in Advanced Melanoma","author":"AJ Olszanski","year":"2014","journal-title":"Journal of Managed Care Pharmacy JMCP April"},{"key":"pone.0281558.ref009","doi-asserted-by":"crossref","first-page":"9","DOI":"10.1007\/s11864-017-0453-5","article-title":"BRAF-Mutated Colorectal Cancer: What Is the Optimal Strategy for Treatment?","volume":"18","author":"R Cohen","year":"2017","journal-title":"Curr Treat Options Oncol"},{"key":"pone.0281558.ref010","doi-asserted-by":"crossref","first-page":"637","DOI":"10.1038\/nrc.2017.84","article-title":"Understanding and targeting resistance mechanisms in NSCLC.","volume":"17","author":"J Rotow","year":"2017","journal-title":"Nat Rev Cancer"},{"key":"pone.0281558.ref011","article-title":"BRAF-mutated colorectal cancer: Clinical and molecular insights","author":"F Caputo","year":"2019","journal-title":"International Journal of Molecular Sciences"},{"key":"pone.0281558.ref012","first-page":"1119","article-title":"Targeting BRAF mutations in non-small cell lung cancer. Translational Lung Cancer Research.","author":"CG O\u2019Leary","year":"2019","journal-title":"AME Publishing Company"},{"key":"pone.0281558.ref013","author":"G Roviello","journal-title":"Advances in anti-BRAF therapies for lung cancer"},{"key":"pone.0281558.ref014","doi-asserted-by":"crossref","first-page":"2507","DOI":"10.1056\/NEJMoa1103782","article-title":"Improved Survival with Vemurafenib in Melanoma with BRAF V600E Mutation.","volume":"364","author":"PB Chapman","year":"2011","journal-title":"New England Journal of Medicine"},{"key":"pone.0281558.ref015","doi-asserted-by":"crossref","first-page":"9043","DOI":"10.1038\/s41598-021-88391-5","article-title":"High sensitivity sanger sequencing detection of BRAF mutations in metastatic melanoma FFPE tissue specimens","volume":"11","author":"LY Cheng","year":"2021","journal-title":"Sci Rep"},{"key":"pone.0281558.ref016","first-page":"660","article-title":"BRAF mutant colorectal cancer as a distinct subset of colorectal cancer: Clinical characteristics, clinical behavior, and response to targeted therapies","volume-title":"Journal of Gastrointestinal Oncology","author":"CN Clarke","year":"2015"},{"key":"pone.0281558.ref017","first-page":"36","article-title":"Clinicopathologic features and treatment efficacy of Chinese patients with BRAF-mutated metastatic colorectal cancer: a retrospective observational study.","author":"X Wang","year":"2017","journal-title":"Chin J Cancer"},{"key":"pone.0281558.ref018","doi-asserted-by":"crossref","first-page":"5705","DOI":"10.1200\/JCO.2008.18.0786","article-title":"Wild-type BRAF is required for response to panitumumab or cetuximab in metastatic colorectal cancer","volume":"26","author":"F di Nicolantonio","year":"2008","journal-title":"Journal of Clinical Oncology"},{"key":"pone.0281558.ref019","first-page":"76","article-title":"Immunohistochemical detection of V600E BRAF mutation is a useful primary screening tool for malignant melanoma","author":"G Orchard","year":"2019","journal-title":"Br J Biomed Sci"},{"key":"pone.0281558.ref020","doi-asserted-by":"crossref","first-page":"157","DOI":"10.2147\/OTT.S39096","article-title":"BRAF-mutant melanoma: Treatment approaches, resistance mechanisms, and diagnostic strategies","author":"F Spagnolo","year":"2015","journal-title":"OncoTargets and Therapy. Dove Medical Press Ltd."},{"key":"pone.0281558.ref021","doi-asserted-by":"crossref","article-title":"Intercalating nucleic acids containing insertions of 1-O-(1-pyrenylmethyl)glycerol: stabilisation of dsDNA and discrimination of DNA over RNA","author":"UB Christensen","DOI":"10.1093\/nar\/gkf624"},{"key":"pone.0281558.ref022","first-page":"12","article-title":"SensiScreen\u00ae KRAS exon 2-sensitive simplex and multiplex real-time PCR-based assays for detection of KRAS exon 2 mutations.","author":"A Riva","year":"2017","journal-title":"PLoS One"},{"key":"pone.0281558.ref023","article-title":"p16 INK4A Hypermethylation Detected by Fluorescent Methylation-specific PCR in Plasmas from Non-Small Cell Lung Cancer 1","author":"A Bearzatto","year":"2002"},{"key":"pone.0281558.ref024","volume-title":"Int J Cancer","author":"P Nollau","year":"1996"},{"key":"pone.0281558.ref025","first-page":"1353","article-title":"Mutations of ras genes distinguish a subset of non-small-cell lung cancer cell lines from small-cell lung cancer cell lines","author":"T Mitsudomi","year":"1991","journal-title":"Oncogene"},{"key":"pone.0281558.ref026","article-title":"Rapid and sensitive nonradioactive detection of mutant K-ras genes via \u201cenriched\u201d PCR amplification.","author":"S Kahn","journal-title":"Oncogene"},{"key":"pone.0281558.ref027","doi-asserted-by":"crossref","first-page":"51","DOI":"10.1016\/0003-2697(91)90293-3","article-title":"A method to detect ras point mutations in small subpopulations of cells","volume":"195","author":"J Chen","year":"1991","journal-title":"Anal Biochem"},{"key":"pone.0281558.ref028","article-title":"NCCN Guidelines Version 2.2022 Colon Cancer Continue NCCN Guidelines Panel Disclosures","author":"AB Benson","year":"2022"}],"container-title":["PLOS ONE"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/dx.plos.org\/10.1371\/journal.pone.0281558","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2023,2,9]],"date-time":"2023-02-09T18:51:01Z","timestamp":1675968661000},"score":1,"resource":{"primary":{"URL":"https:\/\/dx.plos.org\/10.1371\/journal.pone.0281558"}},"subtitle":[],"editor":[{"given":"Alvaro","family":"Galli","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"editor"}]}],"short-title":[],"issued":{"date-parts":[[2023,2,9]]},"references-count":28,"journal-issue":{"issue":"2","published-online":{"date-parts":[[2023,2,9]]}},"URL":"https:\/\/doi.org\/10.1371\/journal.pone.0281558","relation":{},"ISSN":["1932-6203"],"issn-type":[{"value":"1932-6203","type":"electronic"}],"subject":[],"published":{"date-parts":[[2023,2,9]]}}}