Specific Detection of Cytokeratin 20-Positive Cells in Blood of Colorectal and Breast Cancer Patients by a High Sensitivity Real-Time Reverse Transcriptase-Polymerase.

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,

Association of Clinical Status of Follicular Lymphoma Patients after Autologous Stem Cell Transplant and Quantitative Assessment of Lymphoma in Blood and.

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

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

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

Quantitative Detection and Differentiation of Human Herpesvirus 6 Subtypes in Bone Marrow Transplant Patients by Using a Single Real-Time Polymerase Chain.

A Prospective Clinical Study on the Use of Reverse Transcription-Polymerase Chain Reaction for the Early Diagnosis of Dengue Fever Kamaljit Singh, Ajit.

A. Doménech-Sánchez, C. Juan, J.L. Pérez, C.I. Berrocal

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,

by Karen Reue, Robert D. Cohen, and Michael C. Schotz

A. Doménech-Sánchez, C. Juan, J.L. Pérez, C.I. Berrocal

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

Rapid Detection of the Epidermal Growth Factor Receptor Mutation in Non-Small-Cell Lung Cancer for Analysis of Acquired Resistance Using Molecular Beacons

Real-Time Polymerase Chain Reaction for Detecting Bacterial DNA Directly from Blood of Neonates Being Evaluated for Sepsis Jeanne A. Jordan, Mary Beth.

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

Β-Glucuronidase Is an Optimal Normalization Control Gene for Molecular Monitoring of Chronic Myelogenous Leukemia Joong Won Lee, Qiaofang Chen, Daniel.

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

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

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

Philippe Szankasi, Mohamed Jama, David W. Bahler

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,

Association of Clinical Status of Follicular Lymphoma Patients after Autologous Stem Cell Transplant and Quantitative Assessment of Lymphoma in Blood.

Detection of Central Nervous System Leukemia in Children with Acute Lymphoblastic Leukemia by Real-Time Polymerase Chain Reaction Sharon R. Pine, Changhong.

Protocol for the Use of Polymerase Chain Reaction in the Detection of Intraocular Large B-Cell Lymphoma in Ocular Samples Aires Lobo, Narciss Okhravi,

Single Monochrome Real-Time RT-PCR Assay for Identification, Quantification, and Breakpoint Cluster Region Determination of t(9;22) Transcripts Marina.

Single-Run, Parallel Detection of DNA from Three Pneumonia-Producing Bacteria by Real-Time Polymerase Chain Reaction Reinhard B. Raggam, Eva Leitner,

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

A Fast Real-Time Polymerase Chain Reaction Method for Sensitive and Specific Detection of the Neisseria gonorrhoeae porA Pseudogene Stig Ove Hjelmevoll,

Assessment of EGFR Mutation Status in Lung Adenocarcinoma by Immunohistochemistry Using Antibodies Specific to the Two Major Forms of Mutant EGFR Marie.

Rapid and Accurate Detection of Monoclonal Immunoglobulin Heavy Chain Gene Rearrangement by DNA Melting Curve Analysis in the LightCycler System Dongsheng.

Real-Time Polymerase Chain Reaction for Detecting Bacterial DNA Directly from Blood of Neonates Being Evaluated for Sepsis Jeanne A. Jordan, Mary Beth.

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

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

Sensitive Detection of Deletions of One or More Exons in the Neurofibromatosis Type 2 (NF2) Gene by Multiplexed Gene Dosage Polymerase Chain Reaction

Expression of Two Breast-Specific Molecules in the Lung

Analytical Detection of Immunoglobulin Heavy Chain Gene Rearrangements in Gastric Lymphoid Infiltrates by Peak Area Analysis of the Melting Curve in the.

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

Critical Evaluation of Real-Time Reverse Transcriptase-Polymerase Chain Reaction for the Quantitative Detection of Cytokeratin 20 mRNA in Colorectal Cancer.

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 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.

Diagnosis of Trypanosomatid Infections

A Novel Semiquantitative Fluorescence-Based Multiplex Polymerase Chain Reaction Assay for Rapid Simultaneous Detection of Bacterial and Parasitic Pathogens.

Molecular Monitoring of Chronic Myelogenous Leukemia

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

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,

Polymerase Chain Reaction Detection of Kaposi's Sarcoma-Associated Herpesvirus- Optimized Protocols and Their Application to Myeloma Langxing Pan, Laura.

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.

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

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

T Cell Receptor γ-Chain Gene Polymerase Chain Reaction to Diagnose Central Nervous System Involvement by Cutaneous T Cell Lymphoma Robert Taylor, Jo-Anne.

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

Rapid and Accurate Detection of Monoclonal Immunoglobulin Heavy Chain Gene Rearrangement by DNA Melting Curve Analysis in the LightCycler System Dongsheng.

Mangalathu S. Rajeevan, Suzanne D

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

Wook Lew Journal of Investigative Dermatology

Analytical Evaluation of Primer Engineered Multiplex Polymerase Chain Reaction– Restriction Fragment Length Polymorphism for Detection of Factor V Leiden.

Visual Format for Detection of Mycobacterium tuberculosis and M

A Prospective Clinical Study on the Use of Reverse Transcription-Polymerase Chain Reaction for the Early Diagnosis of Dengue Fever Kamaljit Singh, Ajit.

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

Qing-Rong Chen, Gordon Vansant, Kahuku Oades, Maria Pickering, Jun S

Maureen J. O'Sullivan, Peter A. Humphrey, Louis P. Dehner, John D

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:

Specific Detection of Cytokeratin 20-Positive Cells in Blood of Colorectal and Breast Cancer Patients by a High Sensitivity Real-Time Reverse Transcriptase-Polymerase Chain Reaction Method Giuliana Giribaldi, Simone Procida, Daniela Ulliers, Franca Mannu, Roberta Volpatto, Giorgia Mandili, Laura Fanchini, Oscar Bertetto, Gianruggero Fronda, Luigi Simula, Elena Rimini, Giovanni Cherchi, Lisa Bonello, Milena Maria Maule, Francesco Turrini The Journal of Molecular Diagnostics Volume 8, Issue 1, Pages 105-112 (February 2006) DOI: 10.2353/jmoldx.2006.050054 Copyright © 2006 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions

Figure 1 CK20 amplification plots of serially diluted HT29 cells. Serially diluted HT29 cells (1, 104 cells; 2, 103 cells; 3, 102 cells; 4, 101 cells) amplified by S-PCR (A), 20N-PCR (B), and 35N-PCR (C). Numbers of cycles were plotted against fluorescence expressed as relative fluorescence units. The Journal of Molecular Diagnostics 2006 8, 105-112DOI: (10.2353/jmoldx.2006.050054) Copyright © 2006 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions

Figure 2 Melting peak analysis of 20N-PCR products. The melting peaks result from plotting the negative first derivative of measured fluorescence emission (y axis) at a given temperature (x axis). Curves were obtained with pure HT29 cells (A and B); with CK20+ clinical samples (C); with intron-spanning primers E and G (A); and with primer E and exon-overlapping primer F (B and C). The Journal of Molecular Diagnostics 2006 8, 105-112DOI: (10.2353/jmoldx.2006.050054) Copyright © 2006 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions

Figure 3 Nonspecific amplifications by conventional intron spanning primers. Amplification plots obtained with cDNA of lympho-monocytes from healthy donors (A); melting curves obtained from the same PCR reactions (B); gel electrophoresis analysis (2% agarose gel stained with ethidium bromide) of amplified material from lympho-monocytes from healthy donors (controls) and HT29 cells (C). The Journal of Molecular Diagnostics 2006 8, 105-112DOI: (10.2353/jmoldx.2006.050054) Copyright © 2006 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions

Figure 4 Schematic positions of the intron-spanning primers E and G (A) and exon-overlapping primer F (B) used for N-PCR. The Journal of Molecular Diagnostics 2006 8, 105-112DOI: (10.2353/jmoldx.2006.050054) Copyright © 2006 American Society for Investigative Pathology and Association for Molecular Pathology Terms and Conditions