Detection of BRAF p.V600E Mutations in Melanomas

Slides:

Advertisements

Similar presentations

Measurement of Relative Copy Number of CDKN2A/ARF and CDKN2B in Bladder Cancer by Real-Time Quantitative PCR and Multiplex Ligation-Dependent Probe Amplification.

Advertisements

CyclinD1/CyclinD3 Ratio by Real-Time PCR Improves Specificity for the Diagnosis of Mantle Cell Lymphoma Carol D. Jones, Katherine H. Darnell, Roger A.

Clinical Laboratory Analysis of Immunoglobulin Heavy Chain Variable Region Genes for Chronic Lymphocytic Leukemia Prognosis Philippe Szankasi, David.

The Application of Molecular Diagnostics to Stained Cytology Smears

A Virtual Pyrogram Generator to Resolve Complex Pyrosequencing Results

Detection of Low-Level KRAS Mutations Using PNA-Mediated Asymmetric PCR Clamping and Melting Curve Analysis with Unlabeled Probes Ji Eun Oh, Hee Sun.

Laboratory Guidelines for Detection, Interpretation, and Reporting of Maternal Cell Contamination in Prenatal Analyses Narasimhan Nagan, Nicole E. Faulkner,

A Quantitative Allele-Specific PCR Test for the BRAF V600E Mutation Using a Single Heterozygous Control Plasmid for Quantitation Philippe Szankasi, N.

Suppression of Wild-Type Amplification by Selectivity Enhancing Agents in PCR Assays that Utilize SuperSelective Primers for the Detection of Rare Somatic.

Multiplex Preamplification of Serum DNA to Facilitate Reliable Detection of Extremely Rare Cancer Mutations in Circulating DNA by Digital PCR Jennifer.

A Multiplex SNaPshot Assay as a Rapid Method for Detecting KRAS and BRAF Mutations in Advanced Colorectal Cancers Sandrine Magnin, Erika Viel, Alice.

Detection of CALR and MPL Mutations in Low Allelic Burden JAK2 V617F Essential Thrombocythemia Fabrice Usseglio, Nathalie Beaufils, Anne Calleja, Sophie.

A Virtual Pyrogram Generator to Resolve Complex Pyrosequencing Results

Joanna Wang, Chetan Bettegowda The Journal of Molecular Diagnostics

Heterogeneity of BRAF, NRAS, and TERT Promoter Mutational Status in Multiple Melanomas and Association with MC1R Genotype Cristina Pellegrini, Lucia.

Comparison of High-Resolution Melting Analysis, TaqMan Allelic Discrimination Assay, and Sanger Sequencing for Clopidogrel Efficacy Genotyping in Routine.

Detection of BRAF V600 Mutations in Metastatic Melanoma

High prevalence of BRAF V600E mutations in Erdheim-Chester disease but not in other non-Langerhans cell histiocytoses by Julien Haroche, Frédéric Charlotte,

A Novel Targeted Approach for Noninvasive Detection of Paternally Inherited Mutations in Maternal Plasma Jessica M.E. van den Oever, Ivonne J.H.M. van.

Comparison of Allelic Discrimination by dHPLC, HRM, and TaqMan in the Detection of BRAF Mutation V600E Pablo Carbonell, María C. Turpin, Daniel Torres-Moreno,

Frequency and Spectrum of BRAF Mutations in a Retrospective, Single-Institution Study of 1112 Cases of Melanoma Wesley O. Greaves, Shalini Verma, Keyur.

Cornelis J. J. Huijsmans, Jeroen Poodt, Paul H. M. Savelkoul, Mirjam H

Ken B. Waites, Li Xiao, Vanya Paralanov, Rose M. Viscardi, John I

Challenges in Determining Genotypes for Pharmacogenetics in Allogeneic Hematopoietic Cell Transplant Recipients Loralie J. Langman, Lior Nesher, Dimpy.

Molly Yancovitz, Joanne Yoon, Maryann Mikhail, Weiming Gai, Richard L

Detection of KRAS and BRAF Mutations in Colorectal Carcinoma

Assessment of EGFR Mutation Status in Lung Adenocarcinoma by Immunohistochemistry Using Antibodies Specific to the Two Major Forms of Mutant EGFR Marie.

Patrick R. Murray The Journal of Molecular Diagnostics

Christine L. Baker, Cecily P. Vaughn, Wade S. Samowitz

Improved Real-Time Multiplex Polymerase Chain Reaction Detection of Methylenetetrahydrofolate Reductase (MTHFR) 677C>T and 1298A>C Polymorphisms Using.

Verification of Wild-Type EGFR Status in Non–Small Cell Lung Carcinomas Using a Mutant-Enriched PCR on Selected Cases Yi-Lin Chen, Cheng-Chan Lu, Shu-Ching.

Hillary S. Sloane, James P. Landers, Kimberly A. Kelly

Clinical Laboratory Analysis of Immunoglobulin Heavy Chain Variable Region Genes for Chronic Lymphocytic Leukemia Prognosis Philippe Szankasi, David.

Evaluation of BRAF Mutation Testing Methodologies in Formalin-Fixed, Paraffin- Embedded Cutaneous Melanomas Johanne Lade-Keller, Kirsten M. Rømer, Per.

Multiplex Mutation Screening by Mass Spectrometry

Multiplex Amplification Coupled with COLD-PCR and High Resolution Melting Enables Identification of Low-Abundance Mutations in Cancer Samples with Low.

Application of a BRAF Pyrosequencing Assay for Mutation Detection and Copy Number Analysis in Malignant Melanoma Cynthia Spittle, M. Renee Ward, Katherine.

Usefulness of Immunocytochemistry for the Detection of the BRAFV600E Mutation in Circulating Tumor Cells from Metastatic Melanoma Patients Véronique.

KRAS Mutation The Journal of Molecular Diagnostics

BRAF Mutation Testing in Solid Tumors

Development of a Quantitative Real-Time Polymerase Chain Reaction Assay for the Detection of the JAK2 V617F Mutation Elizabeth C. Wolstencroft, Katy.

Highly Sensitive Droplet Digital PCR Method for Detection of EGFR-Activating Mutations in Plasma Cell–Free DNA from Patients with Advanced Non–Small Cell.

Allele-Specific, Non-Extendable Primer Blocker PCR (AS-NEPB-PCR) for DNA Mutation Detection in Cancer Haiying Wang, John Jiang, Bianca Mostert, Anieta.

Ashleigh C. McEvoy, Benjamin A. Wood, Nima M. Ardakani, Michelle R

Novel Heat Pulse Extension-PCR–Based Method for Detection of Large CTG-Repeat Expansions in Myotonic Dystrophy Type 1 Arto K. Orpana, Tho H. Ho, Katariina.

Larissa V. Furtado, Helmut C. Weigelin, Kojo S. J

Detection of BRAF V600E Mutation in Colorectal Cancer

Multiplex Amplification Coupled with COLD-PCR and High Resolution Melting Enables Identification of Low-Abundance Mutations in Cancer Samples with Low.

Larissa V. Furtado, Helmut C. Weigelin, Kojo S. J

An Allele-Specific PCR System for Rapid Detection and Discrimination of the CYP2C19∗4A, ∗4B, and ∗17 Alleles Stuart A. Scott, Qian Tan, Usman Baber,

Design and Evaluation of a Real-Time PCR Assay for Quantification of JAK2 V617F and Wild-Type JAK2 Transcript Levels in the Clinical Laboratory Jason.

Application of COLD-PCR for Improved Detection of NF2 Mosaic Mutations

A Novel Long-Range PCR Sequencing Method for Genetic Analysis of the Entire PKD1 Gene Ying-Cai Tan, Alber Michaeel, Jon Blumenfeld, Stephanie Donahue,

Nested High-Resolution Melting Curve Analysis

Mutation Detection of Epidermal Growth Factor Receptor and KRAS Genes Using the Smart Amplification Process Version 2 from Formalin-Fixed, Paraffin-Embedded.

Rapid and Sensitive Real-Time Polymerase Chain Reaction Method for Detection and Quantification of 3243A>G Mitochondrial Point Mutation Rinki Singh,

Low Incidence of Minor BRAF V600 Mutation-Positive Subclones in Primary and Metastatic Melanoma Determined by Sensitive and Quantitative Real-Time PCR

Next-Generation Genotyping by Digital PCR to Detect and Quantify the BRAF V600E Mutation in Melanoma Biopsies Pierre-Jean Lamy, Florence Castan, Nicolas.

Carrie C. Lubitz, Sareh Parangi, Tammy M. Holm, M

Novel Method for PIK3CA Mutation Analysis

Improved Detection of KIT Exon 11 Duplications in Formalin-Fixed, Paraffin-Embedded Gastrointestinal Stromal Tumors Jerzy Lasota, Bartosz Wasag, Sonja.

Evaluation of High-Resolution Melting Analysis as a Diagnostic Tool to Detect the BRAF V600E Mutation in Colorectal Tumors Martin Pichler, Marija Balic,

Optimized Allele-Specific Real-Time PCR Assays for the Detection of Common Mutations in KRAS and BRAF Alois H. Lang, Heinz Drexel, Simone Geller-Rhomberg,

Two Novel Methods for Rapid Detection and Quantification of DNMT3A R882 Mutations in Acute Myeloid Leukemia Melissa Mancini, Syed Khizer Hasan, Tiziana.

Comparison of Sanger Sequencing, Pyrosequencing, and Melting Curve Analysis for the Detection of KRAS Mutations Athanasios C. Tsiatis, Alexis Norris-Kirby,

Varying Mutational Alterations in Multiple Primary Melanomas

Nathan D. Montgomery, Sara R. Selitsky, Nirali M. Patel, D

Improved Detection of the KIT D816V Mutation in Patients with Systemic Mastocytosis Using a Quantitative and Highly Sensitive Real-Time qPCR Assay Thomas.

Method of Mutation Analysis May Contribute to Discrepancies in Reports of V599EBRAF Mutation Frequencies in Melanocytic Neoplasms Christopher J. Miller,

Optimized Allele-Specific Real-Time PCR Assays for the Detection of Common Mutations in KRAS and BRAF Alois H. Lang, Heinz Drexel, Simone Geller-Rhomberg,

Presentation transcript:

Detection of BRAF p.V600E Mutations in Melanomas Emeline Colomba, Zofia Hélias-Rodzewicz, Andreas Von Deimling, Cristi Marin, Nathalie Terrones, Dominique Pechaud, Sylvie Surel, Jean-François Côté, Frédérique Peschaud, David Capper, Hélène Blons, Ute Zimmermann, Thierry Clerici, Philippe Saiag, Jean- François Emile The Journal of Molecular Diagnostics Volume 15, Issue 1, Pages 94-100 (January 2013) DOI: 10.1016/j.jmoldx.2012.09.001 Copyright © 2013 American Society for Investigative Pathology and the Association for Molecular Pathology Terms and Conditions

Figure 1 Determination of the reference responses by using the results of the four methods. The Journal of Molecular Diagnostics 2013 15, 94-100DOI: (10.1016/j.jmoldx.2012.09.001) Copyright © 2013 American Society for Investigative Pathology and the Association for Molecular Pathology Terms and Conditions

Figure 2 False-negative results with Sanger sequencing performed on samples containing >60% of tumor cells. Left column: Five chromatograms with a negative control (Wild type), three false-negative chromatograms (Y10.1758, Y10.1760, and Y11.0183), and a positive control (p.V600E). Right column: Pyrosequencing (top) and real-time PCR (bottom) of the same cases. The mutant allele (red curve) was only detectable with PNA inhibition of the WT allele. The injection sequence of programs was 5′-GACAACGATGTCAATCTC-3′. The Journal of Molecular Diagnostics 2013 15, 94-100DOI: (10.1016/j.jmoldx.2012.09.001) Copyright © 2013 American Society for Investigative Pathology and the Association for Molecular Pathology Terms and Conditions

Figure 3 Detection of p.V600K with p.V600E-specific real-time PCR. Amplification curves of four cases with a negative control, two p.V600K cases, and a p.V600E-positive control. Without PNAs, amplification of both mutated (red) and WT (green) alleles was obvious on the p.V600E-positive control, whereas only the WT were detectable on the three other samples. The p.V600K mutations (red curves) were only detectable with the PNA inhibition of WT allele amplification. The Journal of Molecular Diagnostics 2013 15, 94-100DOI: (10.1016/j.jmoldx.2012.09.001) Copyright © 2013 American Society for Investigative Pathology and the Association for Molecular Pathology Terms and Conditions

Figure 4 VE1 IHC of melanoma tissue microarray samples. Top to bottom: Two BRAF p.V600E-mutated positive melanomas with a strong cytoplasmic red staining of melanoma cells. Two BRAF WT cases negative for VE1, the first one containing single positively stained macrophages, an artifact easily distinguishable from the positive tumor staining in p.V600E mutated melanomas, and the second one with a high amount of melanin. The bottom case is one of the seven cases that were considered as not interpretable by IHC because of an ambiguous, mostly nuclear staining reaction. The Journal of Molecular Diagnostics 2013 15, 94-100DOI: (10.1016/j.jmoldx.2012.09.001) Copyright © 2013 American Society for Investigative Pathology and the Association for Molecular Pathology Terms and Conditions