| |  Online-Ressource |
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| Verfasst von: | Skorokhod, Alexander [VerfasserIn]  |
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| | Helmbold, Peter [VerfasserIn] |
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| | Brors, Benedikt [VerfasserIn] |
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| | Schirmacher, Peter [VerfasserIn] |
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| | Enk, Alexander [VerfasserIn] |
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| | Penzel, Roland [VerfasserIn] |
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| Titel: | Automated universal BRAF state detection within the activation segment in skin metastases by pyrosequencing-based assay U-BRAFV600 |
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| Verf.angabe: | Alexander Skorokhod, Peter Helmbold, Benedikt Brors, Peter Schirmacher, Alexander Enk, Roland Penzel |
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| Jahr: | 2013 |
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| Umfang: | 10 S. |
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| Fussnoten: | Im Text ist "V600" hochgestellt ; Published: March 26, 2013 ; Gesehen am 03.02.2022 |
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| Titel Quelle: | Enthalten in: PLOS ONE |
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| Ort Quelle: | San Francisco, California, US : PLOS, 2006 |
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| Jahr Quelle: | 2013 |
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| Band/Heft Quelle: | 8(2013), 3, Artikel-ID e59221, Seite 1-10 |
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| ISSN Quelle: | 1932-6203 |
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| Abstract: | Malignant melanoma is a highly-aggressive type of malignancy with considerable metastatic potential and frequent resistance to cytotoxic agents. BRAF mutant protein was recently recognized as therapeutic target in metastatic melanoma. We present a newly-developed U-BRAFV600 approach - a universal pyrosequencing-based assay for mutation detection within activation segment in exon 15 of human braf. We identified 5 different BRAF mutations in a single assay analyzing 75 different formalin-fixed paraffin-embedded (FFPE) samples of cutaneous melanoma metastases from 29 patients. We found BRAF mutations in 21 of 29 metastases. All mutant variants were quantitatively detectable by the newly-developed U-BRAFV600 assay. These results were confirmed by ultra-deep-sequencing validation (∼60,000-fold coverage). In contrast to all other BRAF state detection methods, the U-BRAFV600 assay is capable of automated quantitative identification of at least 36 previously-published BRAF mutations. Under the precaution of a minimum of 3% mutated cells in front of a background of wild type cells, U-BRAFV600 assay design completely excludes false wild-type results. The corresponding algorithm for classification of BRAF-mutated variants is provided. The single-reaction assay and data analysis automation makes our approach suitable for the assessment of large clinical sample sizes. Therefore, we suggest U-BRAFV600 assay as a most powerful sequencing-based diagnostic tool to automatically identify BRAF state as a prerequisite to targeted therapy. |
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| DOI: | doi:10.1371/journal.pone.0059221 |
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| URL: | Bitte beachten Sie: Dies ist ein Bibliographieeintrag. Ein Volltextzugriff für Mitglieder der Universität besteht hier nur, falls für die entsprechende Zeitschrift/den entsprechenden Sammelband ein Abonnement besteht oder es sich um einen OpenAccess-Titel handelt.
kostenfrei: Volltext: https://doi.org/10.1371/journal.pone.0059221 |
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| | kostenfrei: Volltext: https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0059221 |
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| | DOI: https://doi.org/10.1371/journal.pone.0059221 |
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| Datenträger: | Online-Ressource |
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| Sprache: | eng |
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| Sach-SW: | Amplification-refractory mutation system analysis |
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| | Colorectal cancer |
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| | Cutaneous melanoma |
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| | Dideoxy DNA sequencing |
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| | Metastasis |
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| | Mutation databases |
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| | Mutation detection |
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| | Polymerase chain reaction |
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| K10plus-PPN: | 1788479912 |
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| Verknüpfungen: | → Zeitschrift |
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