Comparative Methods to Improve the Detection of BRAF V600 Mutations in Highly Pigmented Melanoma Specimens
| Autor: | Thierry Maudelonde, Eric Frouin, Sylvain Godreuil, Jérôme Solassol, Julie A. Vendrell, Romain Senal, Valérie Costes, Marion Larrieux |
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| Přispěvatelé: | Centre hospitalier universitaire de Poitiers (CHU Poitiers), Service de Biopathologie [CHRU Montpellier], Centre Hospitalier Régional Universitaire [Montpellier] (CHRU Montpellier), Aide à la Décision pour une Médecine Personnalisé - Laboratoire de Biostatistique, Epidémiologie et Recherche Clinique - EA 2415 (AIDMP), Université Montpellier 1 (UM1)-Université de Montpellier (UM), UNICANCER - Institut régional du Cancer Montpellier Val d'Aurelle (ICM), CRLCC Val d'Aurelle - Paul Lamarque, Pathogénèse et contrôle des infections chroniques (PCCI), Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Montpellier (UM)-Centre Hospitalier Universitaire de Montpellier (CHU Montpellier ), Institut de Recherche en Cancérologie de Montpellier (IRCM - U1194 Inserm - UM), CRLCC Val d'Aurelle - Paul Lamarque-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Montpellier (UM), Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre Hospitalier Universitaire de Montpellier (CHU Montpellier )-Université de Montpellier (UM) |
| Jazyk: | angličtina |
| Rok vydání: | 2016 |
| Předmět: |
Male
Pigments Melanomas 0301 basic medicine Skin Neoplasms DNA polymerase Gene Identification and Analysis lcsh:Medicine Skin Pigmentation Artificial Gene Amplification and Extension Polymerase Chain Reaction law.invention chemistry.chemical_compound Sequencing techniques 0302 clinical medicine law Melanin Medicine and Health Sciences DNA sequencing Bovine serum albumin lcsh:Science Melanoma DNA extraction Polymerase chain reaction Multidisciplinary biology Middle Aged 3. Good health Oncology 030220 oncology & carcinogenesis Physical Sciences Female Research Article Proto-Oncogene Proteins B-raf Genotyping Materials Science [SDV.CAN]Life Sciences [q-bio]/Cancer Research and Analysis Methods 03 medical and health sciences Extraction techniques Cell Line Tumor Genetics Humans Molecular Biology Techniques Molecular Biology Mutation Detection Materials by Attribute Aged Organic Pigments lcsh:R Dideoxy DNA sequencing Biology and Life Sciences Cancers and Neoplasms Molecular biology genomic DNA 030104 developmental biology chemistry Mutation biology.protein Nucleic acid lcsh:Q DNA [SDV.MHEP.DERM]Life Sciences [q-bio]/Human health and pathology/Dermatology |
| Zdroj: | PLoS ONE PLoS ONE, Public Library of Science, 2016, 11 (7), pp.e0158698. ⟨10.1371/journal.pone.0158698⟩ PLoS ONE, Vol 11, Iss 7, p e0158698 (2016) |
| ISSN: | 1932-6203 |
| DOI: | 10.1371/journal.pone.0158698⟩ |
| Popis: | International audience; Genotyping BRAF in melanoma samples is often challenging. The presence of melanin greatly interferes with thermostable DNA polymerases and/or nucleic acids in traditional polymerase chain reaction (PCR)-based methods. In the present work, we evaluated three easy-to-use strategies to improve the detection of pigmented DNA refractory to PCR amplification. These pre-PCR processing methods include the addition of bovine serum albumin (BSA), the dilution of DNA, and the purification of DNA using the NucleoSpin® gDNA Clean-up XS Kit. We found that BRAF genotyping in weakly and moderately pigmented samples was more efficient when the sample was processed with BSA or purified with a NucleoSpin® gDNA Clean-up XS Kit prior to PCR amplification. In addition, the combination of both methods resulted in successful detection of BRAF mutation in pigmented specimens, including highly pigmented samples, thereby increasing the chance of patients being elicited for anti-BRAF treatment. These solutions to overcome melanin-induced PCR inhibition are of tremendous value and provide a simple solution for clinical chemistry and routine laboratory medicine. |
| Databáze: | OpenAIRE |
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