Clonality Analysis of B-Cell Lymphoproliferative Disorders Using PCR and Melting Curve Analysis
| Autor: | Larry Dobbs, Lori Earls |
|---|---|
| Rok vydání: | 2003 |
| Předmět: |
Adult
Male Lymphoma B-Cell Gene Rearrangement B-Lymphocyte Heavy Chain Lymphoproliferative disorders Nucleic Acid Denaturation Polymerase Chain Reaction Sensitivity and Specificity Fluorescence Melting curve analysis Pathology and Forensic Medicine Consensus Sequence medicine Humans Child Molecular Biology Polyacrylamide gel electrophoresis B cell Aged DNA Primers Aged 80 and over Clonality Analysis Chemistry Reproducibility of Results DNA Neoplasm Cell Biology Middle Aged Amplicon medicine.disease Molecular biology Clone Cells Lymphoma medicine.anatomical_structure Child Preschool Monoclonal Electrophoresis Polyacrylamide Gel Female |
| Zdroj: | Diagnostic Molecular Pathology. 12:212-223 |
| ISSN: | 1052-9551 |
| DOI: | 10.1097/00019606-200312000-00005 |
| Popis: | Detection of a monoclonal immunoglobulin heavy chain gene (IgH gene) rearrangement is commonly used to support the diagnosis of B-cell non-Hodgkin lymphoma. We investigated the application of melting curve analysis as a substitute for polyacrylamide gel electrophoresis (PAGE) in the detection of monoclonal IgH gene rearrangements after PCR. A total of 140 cases were selected for this study, including 63 B-cell malignancies with a previously documented monoclonal IgH gene rearrangement. These 140 specimens were tested using PCR with melting curve analysis, and the results obtained were compared with PAGE results to calculate the relative sensitivity and specificity of melting curve analysis. Melting curve analysis detected monoclonal rearrangements in 56 of 63 specimens (relative sensitivity 88.9%). No false positives were detected (relative specificity = 100%). False-negative results were obtained only when a weak monoclonal band was present on PAGE. These results show that a positive result on melting curve analysis is specific for a monoclonal IgH gene rearrangement. However, with a sensitivity of only 88.9%, the majority of negative results would require further evaluation of the amplicons using PAGE. The application of melting curve analysis in the detection of monoclonal IgH gene rearrangements in the clinical laboratory setting is discussed. |
| Databáze: | OpenAIRE |
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