Novel Method for PIK3CA Mutation Analysis

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Presentation transcript:

Novel Method for PIK3CA Mutation Analysis Daphne Ang, Rebecca O'Gara, Amy Schilling, Carol Beadling, Andrea Warrick, Megan L. Troxell, Christopher L. Corless The Journal of Molecular Diagnostics Volume 15, Issue 3, Pages 312-318 (May 2013) DOI: 10.1016/j.jmoldx.2012.12.005 Copyright © 2013 American Society for Investigative Pathology and the Association for Molecular Pathology Terms and Conditions

Figure 1 Example of a case of double mutant yielding a false-negative PIK3CA mutation by standard sequencing but a positive result by PCR-MS (Sequenom) and LNA-PCR assay. Left panel: PIK3CA exon 9 mutation E542K (G>A). Right panel: PIK3CA exon 20 mutation H1047R (A>G) (area of mutation shaded in blue in sequence tracings; rightmost peak in Sequenom tracings represents a large wild-type peak). The Journal of Molecular Diagnostics 2013 15, 312-318DOI: (10.1016/j.jmoldx.2012.12.005) Copyright © 2013 American Society for Investigative Pathology and the Association for Molecular Pathology Terms and Conditions

Figure 2 PIK3CA exon 20 sensitivity study: comparison of electropherograms in sequencing with standard Sanger (left column) and LNA-PCR assay (right column). Serial dilutions of mutant DNA with normal control DNA. Sanger sequencing shows a mutant peak at 40% and 20% mutant. Below this level, the mutation is at the level of background or not detectable (area of mutation shaded in blue). With LNA-PCR assay, a mutant peak is evident even at the lowest dilution of 0.6% mutant. The Journal of Molecular Diagnostics 2013 15, 312-318DOI: (10.1016/j.jmoldx.2012.12.005) Copyright © 2013 American Society for Investigative Pathology and the Association for Molecular Pathology Terms and Conditions