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

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Presentation transcript:

Next-Generation Genotyping by Digital PCR to Detect and Quantify the BRAF V600E Mutation in Melanoma Biopsies Pierre-Jean Lamy, Florence Castan, Nicolas Lozano, Cécile Montélion, Patricia Audran, Frédéric Bibeau, Sylvie Roques, Frédéric Montels, Anne-Claire Laberenne The Journal of Molecular Diagnostics Volume 17, Issue 4, Pages 366-373 (July 2015) DOI: 10.1016/j.jmoldx.2015.02.004 Copyright © 2015 American Society for Investigative Pathology and the Association for Molecular Pathology Terms and Conditions

Figure 1 Assessment of the analytical sensitivity of dPCR. Histograms show the fluorescence amplitude (y axis) of droplets after amplification or not by dPCR. Droplets contained serial dilutions of IGR37 genomic DNA (BRAF V600E mutation) in human genomic DNA without BRAF mutations. On the x axis, columns 1, 2: IGR37; 3, 4: IGR37 1/2; 5, 6: IGR37 1/10; 7, 8: IGR371/20; 9, 10: IGR37 1/40; 11, 12: IGR37 1/80; 13, 14: IGR37 1/160; 15, 16: IGR37 1/320; 17, 18: IGR37 1/640; 18, 20: IGR37 1/1280; 21, 22: IGR37 1/2560; 23, 24 IGR37 1/5120. A: Channel 1: FAM–BRAF V600E probe for detection of BRAF V600E-mutated amplicons. B: Channel 2: VIC–wild-type BRAF probe for detection of wild-type BRAF amplicons. dPCR, digital PCR. The Journal of Molecular Diagnostics 2015 17, 366-373DOI: (10.1016/j.jmoldx.2015.02.004) Copyright © 2015 American Society for Investigative Pathology and the Association for Molecular Pathology Terms and Conditions

Figure 2 Comparison of the mutational load quantified by digital PCR and pyrosequencing. Mutational load evaluated by pyrosequencing (x axis) and digital PCR (y axis). The regression slope is equal to 1 and the y-intercept is −0.58. The Spearman's coefficient is 0.9256. Numbers above the circles refer to the sample number. The Journal of Molecular Diagnostics 2015 17, 366-373DOI: (10.1016/j.jmoldx.2015.02.004) Copyright © 2015 American Society for Investigative Pathology and the Association for Molecular Pathology Terms and Conditions