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

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

An Allele-Specific RT-PCR Assay to Detect Type A Mutation of the Nucleophosmin-1 Gene in Acute Myeloid Leukemia Tiziana Ottone, Emanuele Ammatuna, Serena.

Keyur P. Patel, Bedia A. Barkoh, Zhao Chen, Deqin Ma, Neelima Reddy, L

Detection of Exon 12 Mutations in the JAK2 Gene

Mutant Enrichment with 3′-Modified Oligonucleotides

Carol Beadling, Tanaya L. Neff, Michael C

Simultaneous Genotyping of α-Thalassemia Deletional and Nondeletional Mutations by Real-Time PCR–Based Multicolor Melting Curve Analysis Qiuying Huang,

Todd S. Laughlin, Michael W. Becker, Jane L. Liesveld, Deborah A

One-Step Ligation on RNA Amplification for the Detection of Point Mutations Lei Zhang, Jingjing Wang, Mia Coetzer, Stephanie Angione, Rami Kantor, Anubhav.

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

Minimal Residual Disease Monitoring of Acute Myeloid Leukemia by Massively Multiplex Digital PCR in Patients with NPM1 Mutations Nuria Mencia-Trinchant,

Ligation with Nucleic Acid Sequence–Based Amplification

High-Fidelity DNA Polymerase Enhances the Sensitivity of a Peptide Nucleic Acid Clamp PCR Assay for K-ras Mutations Bjørnar Gilje, Reino Heikkilä, Satu.

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

Design and Feasibility of a Novel, Rapid, and Simple Fluorescence 26-Plex RT-PCR Assay for Simultaneous Detection of 24 Fusion Transcripts in Adult Acute.

Isothermal Multiple Displacement Amplification

Establishment and Study of Different Real-Time Polymerase Chain Reaction Assays for the Quantification of Cells with Deletions of Chromosome 7 Elia Mattarucchi,

A DNA Real-Time Quantitative PCR Method Suitable for Routine Monitoring of Low Levels of Minimal Residual Disease in Chronic Myeloid Leukemia Paul A.

Elena Castellanos-Rizaldos, Cloud Paweletz, Chen Song, Geoffrey R

Single Nucleotide Polymorphism-Based System Improves the Applicability of Quantitative PCR for Chimerism Monitoring Egle Gineikiene, Mindaugas Stoskus,

A Locked Nucleic Acid Clamp-Mediated PCR Assay for Detection of a p53 Codon 249 Hotspot Mutation in Urine Selena Y. Lin, Veerpal Dhillon, Surbhi Jain,

A Pyrosequencing-Based Assay for the Rapid Detection of IDH1 Mutations in Clinical Samples Prashanth Setty, Jennifer Hammes, Thomas Rothämel, Valentina.

Tissue-specific Calibration of Real-time PCR Facilitates Absolute Quantification of Plasmid DNA in Biodistribution Studies Joan K Ho, Paul J White, Colin.

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

Hou-Sung Jung, Gregory J. Tsongalis, Joel A. Lefferts

Jeung-Yeal Ahn, Katie Seo, Olga Weinberg, Scott D. Boyd, Daniel A

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

Simultaneous Genotyping of α-Thalassemia Deletional and Nondeletional Mutations by Real-Time PCR–Based Multicolor Melting Curve Analysis Qiuying Huang,

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

Detecting 22q11.2 Deletions by Use of Multiplex Ligation-Dependent Probe Amplification on DNA from Neonatal Dried Blood Spot Samples Karina M. Sørensen,

Detection of an Apparent Homozygous 3120G>A Cystic Fibrosis Mutation on a Routine Carrier Screen Denise LaMarche Heaney, Patrick Flume, Lauren Hamilton,

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

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

Detection of Exon 12 Mutations in the JAK2 Gene

Rapid Detection of TEM-Type Extended-Spectrum β-Lactamase (ESBL) Mutations Using Lights-On/Lights-Off Probes with Single-Stranded DNA Amplification Kenneth.

Keyur P. Patel, Bedia A. Barkoh, Zhao Chen, Deqin Ma, Neelima Reddy, L

Benjamin P. Song, Surbhi Jain, Selena Y. Lin, Quan Chen, Timothy M

A Real-Time Polymerase Chain Reaction Assay for Rapid, Sensitive, and Specific Quantification of the JAK2V617F Mutation Using a Locked Nucleic Acid-Modified.

BRAF Mutation Testing in Solid Tumors

Rapid and Inexpensive Detection of α1-Antitrypsin Deficiency-Related Alleles S and Z by a Real-Time Polymerase Chain Reaction Suitable for a Large-Scale.

A Multi-Site Study for Detection of the Factor V (Leiden) Mutation from Genomic DNA Using a Homogeneous Invader Microtiter Plate Fluorescence Resonance.

Absence of the Caveolin-1 P132L Mutation in Cancers of the Breast and Other Organs Shinya Koike, Yasuhiro Kodera, Akimasa Nakao, Hiroji Iwata, Yasushi.

Detection of FLT3 Internal Tandem Duplication and D835 Mutations by a Multiplex Polymerase Chain Reaction and Capillary Electrophoresis Assay Kathleen.

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.

Development and Laboratory Evaluation of a Real-Time PCR Assay for Detecting Viruses and Bacteria of Relevance for Community-Acquired Pneumonia Alicia.

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

Julie Di Cristofaro, Monique Silvy, Jacques Chiaroni, Pascal Bailly

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

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

Detection of the JAK2 V617F Mutation by LightCycler PCR and Probe Dissociation Analysis Marla Lay, Rajan Mariappan, Jason Gotlib, Lisa Dietz, Siby Sebastian,

Retrospective Comparison of Nucleic Acid Sequence–Based Amplification, Real-Time PCR, and Galactomannan Test for Diagnosis of Invasive Aspergillosis

Development of a High-Resolution Melting Curve Analysis Screening Test for SRSF2 Splicing Factor Gene Mutations in Myelodysplastic Syndromes Eduardo.

Development of an Allele-Specific Real-Time PCR Assay for Discrimination and Quantification of p63 R279H Mutation in EEC Syndrome Vanessa Barbaro, Letizia.

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

An Allele-Specific RT-PCR Assay to Detect Type A Mutation of the Nucleophosmin-1 Gene in Acute Myeloid Leukemia Tiziana Ottone, Emanuele Ammatuna, Serena.

Amplification Refractory Mutation System, a Highly Sensitive and Simple Polymerase Chain Reaction Assay, for the Detection of JAK2 V617F Mutation in Chronic.

Danielle C. Smith, Alina Esterhuizen, Jacquie Greenberg

A Real-Time PCR Assay for the Simultaneous Detection of Functional N284I and L412F Polymorphisms in the Human Toll-Like Receptor 3 Gene Robert A. Brown,

Novel Method for PIK3CA Mutation Analysis

Rapid Polymerase Chain Reaction-Based Detection of Epidermal Growth Factor Receptor Gene Mutations in Lung Adenocarcinomas Qiulu Pan, William Pao, Marc.

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

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,

Custom Design of a GeXP Multiplexed Assay Used to Assess Expression Profiles of Inflammatory Gene Targets in Normal Colon, Polyp, and Tumor Tissue Janice.

Extra Alleles in FMR1 Triple-Primed PCR

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

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,

Quantification of bcl-2/JH Fusion Sequences and a Control Gene by Multiplex Real- Time PCR Coupled with Automated Amplicon Sizing by Capillary Electrophoresis

Presentation transcript:

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 179-184 (March 2015) DOI: 10.1016/j.jmoldx.2014.10.003 Copyright © 2015 American Society for Investigative Pathology and the Association for Molecular Pathology Terms and Conditions

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 2015 17, 179-184DOI: (10.1016/j.jmoldx.2014.10.003) Copyright © 2015 American Society for Investigative Pathology and the Association for Molecular Pathology Terms and Conditions

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 2015 17, 179-184DOI: (10.1016/j.jmoldx.2014.10.003) Copyright © 2015 American Society for Investigative Pathology and the Association for Molecular Pathology Terms and Conditions

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 2015 17, 179-184DOI: (10.1016/j.jmoldx.2014.10.003) Copyright © 2015 American Society for Investigative Pathology and the Association for Molecular Pathology Terms and Conditions