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

Slides:

Advertisements

Similar presentations

Development of a Novel One-Tube Isothermal Reverse Transcription Thermophilic Helicase-Dependent Amplification Platform for Rapid RNA Detection James Goldmeyer,

Advertisements

Clinical Laboratory Analysis of Immunoglobulin Heavy Chain Variable Region Genes for Chronic Lymphocytic Leukemia Prognosis Philippe Szankasi, David.

Detection of Exon 12 Mutations in the JAK2 Gene

Allele-Specific Polymerase Chain Reaction for the Imatinib-Resistant KIT D816V and D816F Mutations in Mastocytosis and Acute Myelogenous Leukemia Christopher.

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.

Locked Nucleic Acids Can Enhance the Analytical Performance of Quantitative Methylation-Specific Polymerase Chain Reaction Karen S. Gustafson The Journal.

Rapid detection of resistance in Mycobacterium tuberculosis: a review discussing molecular approaches D. García de Viedma Clinical Microbiology and.

Suppression of Wild-Type Amplification by Selectivity Enhancing Agents in PCR Assays that Utilize SuperSelective Primers for the Detection of Rare Somatic.

Kirby Siemering, Shehnaaz S. M. Manji, Wendy M

LightCycler Technology in Molecular Diagnostics

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

Comparison of BIOMED-2 Versus Laboratory-Developed Polymerase Chain Reaction Assays for Detecting T-Cell Receptor-γ Gene Rearrangements Keyur P. Patel,

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

Detection of CALR and MPL Mutations in Low Allelic Burden JAK2 V617F Essential Thrombocythemia Fabrice Usseglio, Nathalie Beaufils, Anne Calleja, Sophie.

Yanggu Shi, Sharon F. Terry, Patrick F. Terry, Lionel G

Methylation-Specific Multiplex Ligation-Dependent Probe Amplification Enables a Rapid and Reliable Distinction between Male FMR1 Premutation and Full-Mutation.

Philippe Szankasi, Mohamed Jama, David W. Bahler

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,

Homogeneous Polymerase Chain Reaction Nucleobase Quenching Assay to Detect the 1-kbp Deletion in CLN3 That Causes Batten Disease Paul G. Rothberg, Denia.

The Effect of Primer-Template Mismatches on the Detection and Quantification of Nucleic Acids Using the 5′ Nuclease Assay Ralph Stadhouders, Suzan D.

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

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

DNA Diagnostics by Surface-Bound Melt-Curve Reactions

Long-Range (17.7 kb) Allele-Specific Polymerase Chain Reaction Method for Direct Haplotyping of R117H and IVS-8 Mutations of the Cystic Fibrosis Transmembrane.

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

Quantification of the Mutant CALR Allelic Burden by Digital PCR

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

Rapid Molecular Profiling of Myeloproliferative Neoplasms Using Targeted Exon Resequencing of 86 Genes Involved in JAK-STAT Signaling and Epigenetic Regulation

Rapid Screening Assay for KRAS Mutations by the Modified Smart Amplification Process Kenji Tatsumi, Yasumasa Mitani, Jun Watanabe, Hideki Takakura, Kanako.

Improved Real-Time Multiplex Polymerase Chain Reaction Detection of Methylenetetrahydrofolate Reductase (MTHFR) 677C>T and 1298A>C Polymorphisms Using.

Quantitative Determination of JAK2 V617F by TaqMan

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

Clinical Laboratory Analysis of Immunoglobulin Heavy Chain Variable Region Genes for Chronic Lymphocytic Leukemia Prognosis Philippe Szankasi, David.

Use of Single Nucleotide Polymorphisms (SNP) and Real-Time Polymerase Chain Reaction for Bone Marrow Engraftment Analysis Dwight H. Oliver, Richard E.

Detection of Exon 12 Mutations in the JAK2 Gene

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.

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

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.

Allele-Specific, Non-Extendable Primer Blocker PCR (AS-NEPB-PCR) for DNA Mutation Detection in Cancer Haiying Wang, John Jiang, Bianca Mostert, Anieta.

A Rapid, Sensitive Assay to Detect EGFR Mutation in Small Biopsy Specimens from Lung Cancer Yasushi Yatabe, Toyoaki Hida, Yoshitsugu Horio, Takayuki.

A Commercial Real-Time PCR Kit Provides Greater Sensitivity than Direct Sequencing to Detect KRAS Mutations Bárbara Angulo, Elena García-García, Rebeca.

A Multi-Exonic BRCA1 Deletion Identified in Multiple Families through Single Nucleotide Polymorphism Haplotype Pair Analysis and Gene Amplification with.

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

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

Detection of Common Disease-Causing Mutations in Mitochondrial DNA (Mitochondrial Encephalomyopathy, Lactic Acidosis with Stroke-Like Episodes MTTL

Characterization of Aberrant Melting Peaks in Unlabeled Probe Assays

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.

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

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

Detection of the Activating JAK2 V617F Mutation in Paraffin-Embedded Trephine Bone Marrow Biopsies of Patients with Chronic Myeloproliferative Diseases

Rapid and Sensitive Real-Time Polymerase Chain Reaction Method for Detection and Quantification of 3243A>G Mitochondrial Point Mutation Rinki Singh,

Ayalew Tefferi, Pierre Noel, Curtis A. Hanson

Rebecca L. Margraf, Rong Mao, Carl T. Wittwer

Low Incidence of Minor BRAF V600 Mutation-Positive Subclones in Primary and Metastatic Melanoma Determined by Sensitive and Quantitative Real-Time PCR

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

Cecily P. Vaughn, Elaine Lyon, Wade S. Samowitz

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.

Evaluation of High-Resolution Melting Analysis as a Diagnostic Tool to Detect the BRAF V600E Mutation in Colorectal Tumors Martin Pichler, Marija Balic,

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,

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

Karen Snow-Bailey, Ph.D., 1961–2006

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

Monique A. Johnson, Marvin J. Yoshitomi, C. Sue Richards

Presentation transcript:

A Real-Time Polymerase Chain Reaction Assay for Rapid, Sensitive, and Specific Quantification of the JAK2V617F Mutation Using a Locked Nucleic Acid-Modified Oligonucleotide Barbara Denys, Hakim El Housni, Friedel Nollet, Bruno Verhasselt, Jan Philippé The Journal of Molecular Diagnostics Volume 12, Issue 4, Pages 512-519 (July 2010) DOI: 10.2353/jmoldx.2010.090137 Copyright © 2010 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions

Figure 1 Schematic overview of the mutation-specific LNA-based qPCR reaction. At the left part of the figure, the perfect matching between the hydrolysis probe and the mutated DNA sequence allows annealing of the probe to the target DNA and consequently the Taq polymerase can hydrolyze the probe. In contrast, the single bp mismatch between the LNA oligonucleotide sequence and the mutated DNA sequence prevents specifically its annealing. At the right part of the figure, the LNA oligonucleotide anneals to wild-type target DNA, thereby preventing the detection of wild-type JAK2 sequences. The LNA oligonucleotide thus allows preferential amplification of the mutated DNA if present. Moreover, the mutated specific hydrolysis probe cannot efficiently hybridize to wild-type target because of a single bp mismatch between both. LNA indicates locked nucleic acid. The Journal of Molecular Diagnostics 2010 12, 512-519DOI: (10.2353/jmoldx.2010.090137) Copyright © 2010 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions

Figure 2 Proportion of JAK2V617F allele burden of the 36 JAK2 mutant patients included in this study. Box-and-Whisker presentation of the results from patients with PV (n = 11), ET (n = 14), PMF (n = 8), and post-PV MF (n = 3). The central box represents 25th to 75th percentile, and the middle line the median. The black vertical line extends from the minimum to the maximum value, excluding “outside” and “far out” values. P values are represented for comparisons between all disease entities. PV indicates polycythemia vera; ET, essential thrombocythemia; PMF, primary myelofibrosis; post-PV MF, post-polycythemic myelofibrosis. The Journal of Molecular Diagnostics 2010 12, 512-519DOI: (10.2353/jmoldx.2010.090137) Copyright © 2010 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions