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

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Presentation on theme: "Two Novel Methods for Rapid Detection and Quantification of DNMT3A R882 Mutations in Acute Myeloid Leukemia  Melissa Mancini, Syed Khizer Hasan, Tiziana."— Presentation transcript:

1 Two Novel Methods for Rapid Detection and Quantification of DNMT3A R882 Mutations in Acute Myeloid Leukemia  Melissa Mancini, Syed Khizer Hasan, Tiziana Ottone, Serena Lavorgna, Claudia Ciardi, Daniela F. Angelini, Francesca Agostini, Adriano Venditti, Francesco Lo-Coco  The Journal of Molecular Diagnostics  Volume 17, Issue 2, Pages (March 2015) DOI: /j.jmoldx Copyright © 2015 American Society for Investigative Pathology and the Association for Molecular Pathology Terms and Conditions

2 Figure 1 A schematic representation of a peptide nucleic acid (PNA) real-time PCR technology for monitoring of DNMT3A R882H mutation. A: In the presence of the mutant allele (CAC), the fluorescent probe can bind the DNA sequence competing with the PNA-binding site. B: In the DNMT3A wild-type sample, the PNA binds the specific CGC codon, blocking the amplification. C: A representative plot of DNMT3A R882H and wild-type samples analyzed by a real-time quantitative PCR assay. RFU, relative fluorescent units. The Journal of Molecular Diagnostics  , DOI: ( /j.jmoldx ) Copyright © 2015 American Society for Investigative Pathology and the Association for Molecular Pathology Terms and Conditions

3 Figure 2 A: Diagram of TauI restriction enzyme assay to identify DNMT3A R882H mutation. B: Examples of the DNMT3A R882H assay. Electropherogram on the left site represents a mutated patient with two peaks at 93 and 145 bp, corresponding to the wild-type and mutated allele, respectively. On the right side is an electropherogram of a restriction enzyme assay for DNMT3A wild-type patient. The Journal of Molecular Diagnostics  , DOI: ( /j.jmoldx ) Copyright © 2015 American Society for Investigative Pathology and the Association for Molecular Pathology Terms and Conditions

4 Figure 3 Kinetic evaluation of the DNMT3A R882H expression in 24 patients with acute myeloid leukemia after induction and consolidation therapy. Error bars represent 95% CIs for the median DNMT3A R882H expression at each therapy time point. The Journal of Molecular Diagnostics  , DOI: ( /j.jmoldx ) Copyright © 2015 American Society for Investigative Pathology and the Association for Molecular Pathology Terms and Conditions

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