Detection of clonal T-cell receptor [gamma ] gene rearrangements using fluorescent-based PCR and automated high-resolution capillary electrophoresis
| Autor: | Cindy Tozer, Bernice M. Benoit, Hanna Rennert, Debra G.B. Leonard, Stuart R. Lessin, Vivian Luo, Robert B. Wilson |
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| Rok vydání: | 2001 |
| Předmět: |
Skin Neoplasms
Biopsy T cell Biology Lymphoma T-Cell Polymerase Chain Reaction Sensitivity and Specificity Fluorescence law.invention chemistry.chemical_compound Capillary electrophoresis law medicine Humans Receptor Gene Polyacrylamide gel electrophoresis Polymerase chain reaction DNA Primers Paraffin Embedding Gene Rearrangement gamma-Chain T-Cell Antigen Receptor Genes T-Cell Receptor gamma Electrophoresis Capillary DNA Neoplasm General Medicine Gene rearrangement Molecular biology Clone Cells Lymphoma T-Cell Cutaneous medicine.anatomical_structure chemistry Electrophoresis Polyacrylamide Gel DNA |
| Zdroj: | Molecular Diagnosis. 6:169-179 |
| ISSN: | 1084-8592 |
| Popis: | Analysis of T-cell receptor gamma (TCR gamma) gene rearrangements by PCR is a powerful tool for detecting clonal T-cell populations for the diagnosis of lymphoid neoplasms. We report a method for TCR gamma PCR analysis using capillary electrophoresis (CE).To define the threshold for identification of a predominant monoclonal population within a polyclonal background, we developed a novel objective parameter of the peak height ratio (Rn) of the peak of interest and the average of the two immediate flanking peaks. After evaluation of monoclonal, reactive, and normal T-cell populations, an Rn of 3.0 or greater was determined to be consistent with a monoclonal population, whereas an Rn between 1.9 and 3.0 was considered an intermediate range. This CE method was compared with the standard denaturing gradient gel electrophoresis (DGGE) method using previously evaluated clinical specimens. Eleven of 12 clinical specimens (92%) with a definitive diagnosis of T-cell lymphoma were monoclonal by CE, with 100% concordance with the DGGE method. Of nine specimens morphologically suspicious for T-cell lymphoma, five specimens were positive by CE analysis compared with four specimens by DGGE. In addition, 14 specimens for staging from patients with known T-cell lymphoma were studied using both the CE and DGGE methods, with a concordance of 86%.CE is a powerful and efficient method for analysis of clonality by TCR gamma PCR. |
| Databáze: | OpenAIRE |
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