Real-time t(11;14) and t(14;18) PCR assays provide sensitive and quantitative assessments of minimal residual disease (MRD)
| Autor: | C Reading, R Ramanathan, C J Gerard, James L. Zehnder, E G Hanania, Carol D. Jones, K Olsson |
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| Rok vydání: | 1999 |
| Předmět: |
Cancer Research
Neoplasm Residual Time Factors Molecular Sequence Data Antigens CD34 Bone Marrow Cells Biology Polymerase Chain Reaction Sensitivity and Specificity Translocation Genetic law.invention Antigen law Tumor Cells Cultured TaqMan Humans Single-Blind Method Polymerase chain reaction Chromosomes Human Pair 14 Clinical Trials as Topic Base Sequence Chromosomes Human Pair 11 Lymphoma Non-Hodgkin Bone Marrow Purging Reproducibility of Results Chromosome Breakage Hematology Minimal residual disease Molecular biology Genes bcl-1 Bone marrow purging Genes bcl-2 Oncology T cell differentiation Thy-1 Antigens Chromosomes Human Pair 18 Nested polymerase chain reaction Ex vivo |
| Zdroj: | Leukemia. 13:1833-1842 |
| ISSN: | 1476-5551 0887-6924 |
| DOI: | 10.1038/sj.leu.2401575 |
| Popis: | Non-Hodgkin's lymphoma (NHL) arises as a clonal transformation of normal B and T cell differentiation and is often characterized by a higher incidence of specific chromosomal translocations. We have developed real-time TaqMan PCR assays directed toward two of these tumor-associated DNA markers, the t(14;18)(q32;q21.3) at the major breakpoint region of the bcl-2 gene and the t(11;14)(q13;q32) at the bcl-1 major translocation cluster. During analysis of serial dilutions of t(14;18)-positive DNA, the t(14;18) real-time PCR was at least as sensitive as nested PCR and demonstrated enhanced quantitative potential. Moreover, in a blinded comparison of the t(14;18) real-time PCR and a clinically validated nested PCR protocol using 134 cell line and patient DNA samples, the real-time PCR detected the translocation in 30.0% more cases than nested PCR. Both the t(14;18) and t(11;14) real-time PCR assays were used to quantitate minimal residual disease (MRD) in an NHL clinical trial assessing the safety and efficacy of a tumor-purging protocol in autologous stem cell transplantation. The assays were also used to evaluate disease depletion in an ex vivo tumor spiking model in which normal peripheral blood was spiked with tumor cell lines and processed according to the clinical purging method. PCR data from both the clinical trial and the ex vivo model demonstrated a 4 to 6 log reduction in tumor cells during CD34+ and CD34+ Thy-1+ enrichment. Because the t(14;18) and t(11;14) real-time PCR assays are very sensitive, quantitative, rapid, and require no post-PCR manipulation, they may serve as practical alternatives to nested PCR. |
| Databáze: | OpenAIRE |
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