Quantitative Determination of JAK2 V617F by TaqMan

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

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

One Hundred Twenty-One Dystrophin Point Mutations Detected from Stored DNA Samples by Combinatorial Denaturing High-Performance Liquid Chromatography Annalaura.

CyclinD1/CyclinD3 Ratio by Real-Time PCR Improves Specificity for the Diagnosis of Mantle Cell Lymphoma Carol D. Jones, Katherine H. Darnell, Roger A.

Margaret L. Gulley, Thomas C. Shea, Yuri Fedoriw

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

Clinical Validation of a New Triplex Real-Time Polymerase Chain Reaction Assay for the Detection and Discrimination of Herpes simplex Virus Types 1 and.

EGFR Mutations in Lung Adenocarcinomas

Detection of Exon 12 Mutations in the JAK2 Gene

Direct DNA Amplification from Crude Clinical Samples Using a PCR Enhancer Cocktail and Novel Mutants of Taq Zhian Zhang, Milko B. Kermekchiev, Wayne.

Evaluation of the Epstein-Barr Virus R-Gene Quantification Kit in Whole Blood with Different Extraction Methods and PCR Platforms Samira Fafi-Kremer,

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

Direct DNA Amplification from Crude Clinical Samples Using a PCR Enhancer Cocktail and Novel Mutants of Taq Zhian Zhang, Milko B. Kermekchiev, Wayne.

Margaret L. Gulley, Thomas C. Shea, Yuri Fedoriw

Performance Characteristics of a Quantitative Hepatitis C Virus RNA Assay Using COBAS AmpliPrep Total Nucleic Acid Isolation and COBAS TaqMan Hepatitis.

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

Comprehensive Analysis of CBFβ-MYH11 Fusion Transcripts in Acute Myeloid Leukemia by RT-PCR Analysis ShriHari S. Kadkol, Annette Bruno, Carol Dodge,

Clinical Validation of a New Triplex Real-Time Polymerase Chain Reaction Assay for the Detection and Discrimination of Herpes simplex Virus Types 1 and.

Molecular Monitoring of Residual Disease in Chronic Myeloid Leukemia by Genomic DNA Compared with Conventional mRNA Analysis Elia Mattarucchi, Orietta.

Mutation Screening in Juvenile Polyposis Syndrome

Detection of Cytomegalovirus in Whole Blood Using Three Different Real-Time PCR Chemistries Erica Vincent, Zhengming Gu, Markus Morgenstern, Candace.

Joanna Wang, Chetan Bettegowda The Journal of Molecular Diagnostics

Limitations and Practical Procedure in BclII-Ig Heavy Chain Gene Rearrangement Real- Time Quantitative Polymerase Chain Reaction Barbara Dessars, Pierre.

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

Evaluation of the Cepheid GeneXpert BCR-ABL Assay

Molecular Monitoring of Residual Disease in Chronic Myeloid Leukemia by Genomic DNA Compared with Conventional mRNA Analysis Elia Mattarucchi, Orietta.

Rapid Real-Time PCR Assays for Detection of Klebsiella pneumoniae with the rmpA or magA Genes Associated with the Hypermucoviscosity Phenotype Laurie.

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

High Resolution Melting Analysis for JAK2 Exon 14 and Exon 12 Mutations Inmaculada Rapado, Silvia Grande, Enriqueta Albizua, Rosa Ayala, José-Angel Hernández,

Patrick R. Murray The Journal of Molecular Diagnostics

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

William L. Gerald, M.D., Ph.D, 1954–2008

Assessing Heteroplasmic Load in Leber's Hereditary Optic Neuropathy Mutation 3460G→A/MT-ND1 with A Real-Time PCR Quantitative Approach Anna Genasetti,

Rapid Detection of Clonal T-Cell Receptor-β Gene Rearrangements in T-Cell Lymphomas Using the LightCycler-Polymerase Chain Reaction with DNA Melting Curve.

Multiplexed Detection of Anthrax-Related Toxin Genes

Kyong-Ah Yoon, Sohee Park, Sang Hee Lee, Jin Hee Kim, Jin Soo Lee

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

The Human Androgen Receptor X-Chromosome Inactivation Assay for Clonality Diagnostics of Natural Killer Cell Proliferations Michaël Boudewijns, Jacques.

Detection of Exon 12 Mutations in the JAK2 Gene

Real-Time Polymerase Chain Reaction Detection of Herpes Simplex Virus in Cerebrospinal Fluid and Cost Savings from Earlier Hospital Discharge Kenneth.

Application of a BRAF Pyrosequencing Assay for Mutation Detection and Copy Number Analysis in Malignant Melanoma Cynthia Spittle, M. Renee Ward, Katherine.

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

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.

EGFR Mutations in Lung Adenocarcinomas

Testing for Maternal Cell Contamination in Prenatal Samples

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

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,

Accurate Detection of Copy Number Changes in DNA Extracted from Formalin-Fixed, Paraffin-Embedded Melanoma Tissue Using Duplex Ratio Tests David A. Moore,

Sensitivity of the ViroSeq HIV-1 Genotyping System for Detection of the K103N Resistance Mutation in HIV-1 Subtypes A, C, and D Jessica D. Church, Dana.

Diagnosis of Human Congenital Cytomegalovirus Infection by Amplification of Viral DNA from Dried Blood Spots on Perinatal Cards Lori Scanga, Shu Chaing,

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

DNA and RNA References for qRT-PCR Assays in Exfoliated Cervical Cells

Performance and Clinical Evaluation of a Sensitive Multiplex Assay for the Rapid Detection of Common NPM1 Mutations Michael Hafez, Fei Ye, Keith Jackson,

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

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

Multiple Sequence Variants in Hereditary Hemorrhagic Telangiectasia Cases Jamie McDonald, Friederike Gedge, Allene Burdette, James Carlisle, Changkuoth.

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

Rapid Identification of Promoter Hypermethylation in Hepatocellular Carcinoma by Pyrosequencing of Etiologically Homogeneous Sample Pools Emelyne Dejeux,

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

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

Karen Snow-Bailey, Ph.D., 1961–2006

Comparison of Sanger Sequencing, Pyrosequencing, and Melting Curve Analysis for the Detection of KRAS Mutations Athanasios C. Tsiatis, Alexis Norris-Kirby,

Detection of BRCA1 and BRCA2 Ashkenazi Jewish Founder Mutations in Formalin- Fixed Paraffin-Embedded Tissues Using Conventional PCR and Heteroduplex/Amplicon.

Validation of Roche LightCycler Epstein-Barr Virus Quantification Reagents in a Clinical Laboratory Setting Margaret L. Gulley, Hongxin Fan, Sandra H.

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

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:

Quantitative Determination of JAK2 V617F by TaqMan Emma Hammond, Kathryn Shaw, Benedict Carnley, Stephanie P'ng, Ian James, Richard Herrmann The Journal of Molecular Diagnostics Volume 9, Issue 2, Pages 242-248 (April 2007) DOI: 10.2353/jmoldx.2007.060125 Copyright © 2007 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions

Figure 1 a: Specificity versus sensitivity. The assay demonstrated sensitivity to 0.05% clinical dilution. Determining the specificity of the assay is hampered by the fact that it is not known whether healthy controls carry low levels of the mutation. However, we can say that amplification of the mutation in controls occurred only after the number of cycles required to detect a 0.05% dilution of the positive sample. b: Measurement of JAK2 V617F copies per reaction in mixtures of DNA from two healthy controls demonstrated a poor relationship between crossing threshold values and dilutions of DNA in water (right) and in DNA from other control sample (left), suggesting nonspecific amplification. The Journal of Molecular Diagnostics 2007 9, 242-248DOI: (10.2353/jmoldx.2007.060125) Copyright © 2007 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions

Figure 2 Distribution of JAK2 V617F copies per cell among cases and controls. The Journal of Molecular Diagnostics 2007 9, 242-248DOI: (10.2353/jmoldx.2007.060125) Copyright © 2007 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions

Figure 3 Relationship between mutational load and disease phenotype. In terms of a pathogenic model, our data support a significant relationship between mutational load and the different disease phenotypes observed in MPD. Our data have also demonstrated a significant positive relationship between mutational load and myeloproliferative markers and PRV-1 overexpression. Other studies have indicated that the mutation load is also correlated with increases in a number of activation markers of myeloid cells, although exactly how these cellular activations relate to thrombo-hemorrhagic and fibrotic complications is still unclear. The Journal of Molecular Diagnostics 2007 9, 242-248DOI: (10.2353/jmoldx.2007.060125) Copyright © 2007 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions