T cell receptor gamma gene rearrangements as targets for detection of minimal residual disease in acute lymphoblastic leukemia by real-time quantitative PCR analysis
| Autor: | Jjm van Dongen, Vhj van der Velden, J M Wijkhuijs, Dch Jacobs, E. R. Van Wering |
|---|---|
| Přispěvatelé: | Immunology |
| Rok vydání: | 2002 |
| Předmět: |
Cancer Research
Neoplasm Residual Touchdown polymerase chain reaction Biology Polymerase Chain Reaction Sensitivity and Specificity law.invention Predictive Value of Tests law Acute lymphocytic leukemia TaqMan medicine Humans Childhood Acute Lymphoblastic Leukemia Polymerase chain reaction Acute leukemia Gene Rearrangement gamma-Chain T-Cell Antigen Receptor Genes T-Cell Receptor gamma Hematology Gene rearrangement Precursor Cell Lymphoblastic Leukemia-Lymphoma medicine.disease Molecular biology Minimal residual disease Oncology Immunology |
| Zdroj: | Leukemia, 16, 1372-1380. Nature Publishing Group |
| ISSN: | 1476-5551 0887-6924 |
| DOI: | 10.1038/sj.leu.2402515 |
| Popis: | Several studies have shown that quantitative detection of minimal residual disease (MRD) predicts clinical outcome in childhood acute lymphoblastic leukemia (ALL). In this report we investigated the applicablility of T cell receptor gamma (TCRG) gene rearrangements as targets for MRD detection by real-time quantitative PCR analysis. Seventeen children with precursor-B-ALL and 15 children with T-ALL were included in this study. Using an allele-specific (ASO) forward primer in combination with germline Jgamma reverse primers and Jgamma TaqMan probes, a reproducible sensitivity ofor =10(-4) (defined by strict criteria) was obtained in only four out of 19 (21%) TCRG gene rearrangements in precursor-B-ALL patients and in 10 out of 15 (67%) TCRG gene rearrangements in T-ALL patients. The main reason for not obtaining a reproducible sensitivity ofor =10(-4) in approximately 60% of cases was the non-specific amplification of TCRG gene rearrangements in normal T-lymphocytes. A maximal sensitivity ofor =10(-4) (defined by less strict criteria) was obtained in 42% of TCRG gene rearrangements in precursor-B-ALL patients. The number of inserted nucleotides was significantly higher in T-ALL (mean: 8.5) as compared to precursor-B-ALL (mean: 6.8) and appeared to be the most important predictor for reaching a reproducible sensitivityor =10(-4). The usage of a touchdown PCR or the usage of an ASO reverse primer in combination with Vgamma member forward primers and TaqMan probes did not clearly improve the overall results. Nevertheless, RQ-PCR analysis of TCRG gene rearrangements in follow-up samples obtained from 12 ALL patients showed the applicability of this method for MRD detection. We conclude that RQ-PCR analysis of TCRG gene rearrangements can be used for the detection of MRD, but that sensitivities might be limited due to non-specific amplification. This method is applicable in the majority of T-ALL patients and in almost half of precursor-B-ALL patients, particularly when used as second-choice target for confirmation of the MRD results obtained via the first-choice target. |
| Databáze: | OpenAIRE |
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