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

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Presentation transcript:

Comparison of Allelic Discrimination by dHPLC, HRM, and TaqMan in the Detection of BRAF Mutation V600E Pablo Carbonell, María C. Turpin, Daniel Torres-Moreno, Irene Molina-Martínez, José García-Solano, Miguel Perez-Guillermo, Pablo Conesa-Zamora The Journal of Molecular Diagnostics Volume 13, Issue 5, Pages 467-473 (September 2011) DOI: 10.1016/j.jmoldx.2011.03.009 Copyright © 2011 American Society for Investigative Pathology and the Association for Molecular Pathology Terms and Conditions

Figure 1 HRM analysis curves of specimens harboring heterozygous V600E (asterisk), WT V600 (empty arrow), and heterozygous R603X (solid arrow) detected by HRM as a non-V600E variant. RFU indicates relative fluorescence units. The Journal of Molecular Diagnostics 2011 13, 467-473DOI: (10.1016/j.jmoldx.2011.03.009) Copyright © 2011 American Society for Investigative Pathology and the Association for Molecular Pathology Terms and Conditions

Figure 2 dHPLC curves obtained for different non-V600E mutations compared with the WT curve (arrows). The Journal of Molecular Diagnostics 2011 13, 467-473DOI: (10.1016/j.jmoldx.2011.03.009) Copyright © 2011 American Society for Investigative Pathology and the Association for Molecular Pathology Terms and Conditions

Figure 3 Comparison of the percentage of cases rendering no amplification for TaqMan (TM), dHPLC, HRM, and sequencing (Seq) techniques, according to the age of paraffin-embedded tissue. The Journal of Molecular Diagnostics 2011 13, 467-473DOI: (10.1016/j.jmoldx.2011.03.009) Copyright © 2011 American Society for Investigative Pathology and the Association for Molecular Pathology Terms and Conditions

Figure 4 Sensitivity test for the detection of the V600E mutation by dHPLC (A) and HRM (B). DNA from the V600E heterozygote cell line was serial diluted in DNA from the WT cell line. Percentages indicate the proportion of the V600E allele. The Journal of Molecular Diagnostics 2011 13, 467-473DOI: (10.1016/j.jmoldx.2011.03.009) Copyright © 2011 American Society for Investigative Pathology and the Association for Molecular Pathology Terms and Conditions

Figure 5 Estimation of minimum cancer content harboring the V600E mutation by dHPLC and HRM. Percentages indicated are the estimated proportion of the V600E allele versus WT in the DNA extracted from paraffin-embedded tissue, as measured by dHPLC (A) and HRM (B). RFU indicates relative fluorescence units. The Journal of Molecular Diagnostics 2011 13, 467-473DOI: (10.1016/j.jmoldx.2011.03.009) Copyright © 2011 American Society for Investigative Pathology and the Association for Molecular Pathology Terms and Conditions