Validation of High-Resolution DNA Melting Analysis for Mutation Scanning of the Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) Gene Marie-Pierre.

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

Design and Multiseries Validation of a Web-Based Gene Expression Assay for Predicting Breast Cancer Recurrence and Patient Survival Ryan K. Van Laar The.

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

Detection of Exon 12 Mutations in the JAK2 Gene

The Use of COLD-PCR and High-Resolution Melting Analysis Improves the Limit of Detection of KRAS and BRAF Mutations in Colorectal Cancer Irene Mancini,

Rapid Molecular Analysis of the STAT3 Gene in Job Syndrome of Hyper-IgE and Recurrent Infectious Diseases Attila Kumánovics, Carl T. Wittwer, Robert.

Cystic Fibrosis The Journal of Molecular Diagnostics

LightCycler Technology in Molecular Diagnostics

A Multiplex SNaPshot Assay as a Rapid Method for Detecting KRAS and BRAF Mutations in Advanced Colorectal Cancers Sandrine Magnin, Erika Viel, Alice.

Mutation Screening in Juvenile Polyposis Syndrome

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

Simultaneous Genotyping of GSTT1 and GSTM1 Null Polymorphisms by Melting Curve Analysis in Presence of SYBR Green I Fátima Marín, Nadia García, Xavier.

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

A High-Resolution Melting Protocol for Rapid and Accurate Differential Diagnosis of Thyroid Nodules Irene Mancini, Pamela Pinzani, Cinzia Pupilli, Luisa.

Comparison of High-Resolution Melting Analysis, TaqMan Allelic Discrimination Assay, and Sanger Sequencing for Clopidogrel Efficacy Genotyping in Routine.

RET Proto-Oncogene Genotyping Using Unlabeled Probes, the Masking Technique, and Amplicon High-Resolution Melting Analysis Rebecca L. Margraf, Rong Mao,

Nucleotide Extension Genotyping by High-Resolution Melting

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.

Influence of the Duplication of CFTR Exon 9 and Its Flanking Sequences on Diagnosis of Cystic Fibrosis Mutations Ayman El-Seedy, Tony Dudognon, Frédéric.

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

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

Quantitative Analyses of SMN1 and SMN2 Based on Real-Time LightCycler PCR: Fast and Highly Reliable Carrier Testing and Prediction of Severity of Spinal.

Influenza A Subtyping The Journal of Molecular Diagnostics

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

Gene Scanning of an Internalin B Gene Fragment Using High-Resolution Melting Curve Analysis as a Tool for Rapid Typing of Listeria monocytogenes Ariane.

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

Comprehensive and Rapid Genotyping of Mutations and Haplotypes in Congenital Bilateral Absence of the Vas Deferens and Other Cystic Fibrosis Transmembrane.

Andrea Gaedigk, Amanda K. Riffel, J. Steven Leeder

Design and Validation of a Conformation Sensitive Capillary Electrophoresis-Based Mutation Scanning System and Automated Data Analysis of the More than.

Multiplex Ligation-Dependent Probe Amplification Versus Multiprobe Fluorescence in Situ Hybridization To Detect Genomic Aberrations in Chronic Lymphocytic.

Detection of Exon 12 Mutations in the JAK2 Gene

Genotyping of Frequent BRCA1/2 SNPs with Unlabeled Probes

Multiplex Amplification Coupled with COLD-PCR and High Resolution Melting Enables Identification of Low-Abundance Mutations in Cancer Samples with Low.

Harry R. Hill, Nancy H. Augustine, Robert J. Pryor, Gudrun H

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

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

A Synonymous Mutation in the CFTR Gene Causes Aberrant Splicing in an Italian Patient Affected by a Mild Form of Cystic Fibrosis Valeria Faa′, Alessandra.

Development of a Quantitative Real-Time Polymerase Chain Reaction Assay for the Detection of the JAK2 V617F Mutation Elizabeth C. Wolstencroft, Katy.

WGA Allows the Molecular Characterization of a Novel Large CFTR Rearrangement in a Black South African Cystic Fibrosis Patient Marie des Georges, Caroline.

The Suitability of Matrix Assisted Laser Desorption/Ionization Time of Flight Mass Spectrometry in a Laboratory Developed Test Using Cystic Fibrosis Carrier.

Detection and Discrimination between Deletional and Non-Deletional Prader-Willi and Angelman Syndromes by Methylation-Specific PCR and Quantitative Melting.

Design, Development, Validation, and Use of Synthetic Nucleic Acid Controls for Diagnostic Purposes and Application to Cystic Fibrosis Testing Todd M.

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

Detection of BRAF V600E Mutation in Colorectal Cancer

Multiplex Amplification Coupled with COLD-PCR and High Resolution Melting Enables Identification of Low-Abundance Mutations in Cancer Samples with Low.

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

Rapid and Reliable Detection of Glucose-6-Phosphate Dehydrogenase (G6PD) Gene Mutations in Han Chinese Using High-Resolution Melting Analysis Jing-bin.

Application of COLD-PCR for Improved Detection of NF2 Mosaic Mutations

A Novel Method for Creating Artificial Mutant Samples for Performance Evaluation and Quality Control in Clinical Molecular Genetics Michael Jarvis, Ramaswamy.

Simultaneous Amplification, Detection, and Analysis of Common Mutations in the Galactose-1-Phosphate Uridyl Transferase Gene Mohamed Jama, Lesa Nelson,

Maternal Uniparental Meroisodisomy in the LAMB3 Region of Chromosome 1 Results in Lethal Junctional Epidermolysis Bullosa Yasuko Takizawa, Leena Pulkkinen,

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

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

Validation of Denaturing High Performance Liquid Chromatography as a Rapid Detection Method for the Identification of Human INK4A Gene Mutations Irene.

Rapid Mutation Screening for HRPT2 and MEN1 Mutations Associated with Familial and Sporadic Primary Hyperparathyroidism Viive M. Howell, John W. Cardinal,

Multiplex PCR Detection of GSTM1, GSTT1, and GSTP1 Gene Variants

Feras M. Hantash, Arlene Rebuyon, Mei Peng, Joy B

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,

X-Linked Anhidrotic (Hypohidrotic) Ectodermal Dysplasia Caused by a Novel Mutation in EDA1 Gene: 406T>G (Leu55Arg) Francisco Martínez, José María Millán,

A New Insertion/Deletion of the Cystic Fibrosis Transmembrane Conductance Regulator Gene Accounts for 3.4% of Cystic Fibrosis Mutations in Sardinia: Implications.

Rapid Allelic Discrimination by TaqMan PCR for the Detection of the Gilbert's Syndrome Marker UGT1A1*28 Ursula Ehmer, Tim O. Lankisch, Thomas J. Erichsen,

Molecular Diagnosis of Mast Cell Disorders

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

Characterization of a Recurrent Novel Large Duplication in the Cystic Fibrosis Transmembrane Conductance Regulator Gene Feras M. Hantash, Joy B. Redman,

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

Multiplex Ligation-Dependent Probe Amplification Identification of Whole Exon and Single Nucleotide Deletions in the CFTR Gene of Hispanic Individuals.

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

Lung Adenocarcinoma Harboring Mutations in the ERBB2 Kinase Domain

Identification of Cystic Fibrosis Variants by Polymerase Chain Reaction/Oligonucleotide Ligation Assay Karen M. Schwartz, Lisa L. Pike-Buchanan, Kasinathan.

Presentation transcript:

Validation of High-Resolution DNA Melting Analysis for Mutation Scanning of the Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) Gene Marie-Pierre Audrezet, Aurélia Dabricot, Cédric Le Marechal, Claude Ferec The Journal of Molecular Diagnostics Volume 10, Issue 5, Pages 424-434 (September 2008) DOI: 10.2353/jmoldx.2008.080056 Copyright © 2008 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions

Figure 1 Melting profiles of different mutations of exon 8: 1249-5A>G, L375F, 1259insA, T388M, and W401X. The Journal of Molecular Diagnostics 2008 10, 424-434DOI: (10.2353/jmoldx.2008.080056) Copyright © 2008 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions

Figure 2 Difference plot of samples carrying five different variants in exon 21. Different melting profiles are obtained for samples carrying different variants and reproducible melting profiles are obtained with different samples carrying the same mutation, or mutations located in the same codon. The Journal of Molecular Diagnostics 2008 10, 424-434DOI: (10.2353/jmoldx.2008.080056) Copyright © 2008 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions

Figure 3 HRM analysis data for exon 14a, which contains the frequent polymorphism T854T. The figure shows that the difference plot profile is modified when another variant (I853I) is associated with this frequent polymorphism. The Journal of Molecular Diagnostics 2008 10, 424-434DOI: (10.2353/jmoldx.2008.080056) Copyright © 2008 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions

Figure 4 HRM analysis data for exon 10, which contains the frequent mutation F508del. Differences in melting plots obtained for a heterozygous F508del and two compounds heterozygous (F508del/S492F and F508del/S489X). The Journal of Molecular Diagnostics 2008 10, 424-434DOI: (10.2353/jmoldx.2008.080056) Copyright © 2008 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions

Figure 5 Some homozygous variants detected: 394delT in exon 3, 1001+11C>T in exon 6b, G542X in exon 11, and 4016insT in exon 21. The Journal of Molecular Diagnostics 2008 10, 424-434DOI: (10.2353/jmoldx.2008.080056) Copyright © 2008 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions

Figure 6 Analysis of a homozygous F508del sample. Difference plots obtained with or without mixing with a normal DNA before PCR (a) and after a first melting (b). a: DNA from a CF patient with analyses alone or after mixing with a known wild-type sample before amplification. The profile obtained for the mixed DNA corresponds to those of a heterozygous sample. b: The samples may also be first analyzed without mixing, and when the result is negative for DNA from CF patients, the PCR product can be mixed with that of a normal DNA. After a denaturation/hybridization cycle of heating at 94°C followed by a rapid cooling to 45°C to generate heteroduplexes, a melting profile corresponding with those of a heterozygous sample is obtained. The Journal of Molecular Diagnostics 2008 10, 424-434DOI: (10.2353/jmoldx.2008.080056) Copyright © 2008 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions

Figure 7 Detection of three common mutations in exon 7. The 1078delT mutation, which is frequent in Brittany shows a subtle curve modification and must be included as a control for the systematic analysis of exon 7. The Journal of Molecular Diagnostics 2008 10, 424-434DOI: (10.2353/jmoldx.2008.080056) Copyright © 2008 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions

Figure 8 HRM analysis data for exon 17a. Because of its size and of the presence of many melting domains, it was split into two overlapping fragments. Part A contains codons 1047 to 1088 and part B contains codons 1082 to 1122. The Journal of Molecular Diagnostics 2008 10, 424-434DOI: (10.2353/jmoldx.2008.080056) Copyright © 2008 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions