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P. Temperani P. Vaccari F. Giacobbi G. Gandini M. G. Ferrari G. Emilia 《European journal of haematology》1995,55(1):10-13
Abstract: To detect the minimal residual disease (MRD) in acute promyelocytic leukaemia patients treated with all-trans retinoic acid, we compared the sensitivity of metaphase fluorescence in situ hybridization (FISH) with conventional analysis of G-banded metaphases. 5 out of 6 patients studied at diagnosis showed the t(15;17) translocation. 4 out of 5 patients carrying t(15;17) achieved complete remission and conventional cytogenetic conversion. In 3 cases the whole chromosome painting (WCP) probe 17 discovered one normal chromosome 17 and two fragments indicative of t(15;17) persistence. The FISH-WCP technique seems to be highly sensitive and recommendable in monitoring leukaemias with specific chromosome rearrangements. 相似文献
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Localization of the G-CSF gene on chromosome 17 proximal to the breakpoint in the t(15;17) in acute promyelocytic leukemia 总被引:3,自引:0,他引:3
Simmers RN; Webber LM; Shannon MF; Garson OM; Wong G; Vadas MA; Sutherland GR 《Blood》1987,70(1):330-332
The human granulocyte-colony stimulating factor gene (G-CSF) is localized at 17q11.2-q21, the region of one of the breakpoints in the 15;17 chromosome translocation specific for acute promyelocytic leukemia (APL). As G-CSF induces differentiation and loss of tumorigenicity in myeloid leukemic cells or cell lines, it was possible that the translocation in APL involved the DNA of the G-CSF coding region or its regulatory region. In situ hybridization to chromosomes with the t(15;17) from patients with the APL translocation using a G- CSF cDNA clone revealed that the coding region of this gene is proximal to the t(15;17) breakpoint on chromosome 17. Southern analysis of DNA from patients with the APL translocation showed no differences in hybridization between normal and leukemic cells. These results indicate that the G-CSF coding sequence is not disrupted by the chromosomal rearrangement characteristic of APL. 相似文献
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Reverse transcription polymerase chain reaction for the rearranged retinoic acid receptor alpha clarifies diagnosis and detects minimal residual disease in acute promyelocytic leukemia. 总被引:5,自引:0,他引:5
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W H Miller Jr A Kakizuka S R Frankel R P Warrell Jr A DeBlasio K Levine R M Evans E Dmitrovsky 《Proceedings of the National Academy of Sciences of the United States of America》1992,89(7):2694-2698
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Detection of minimal residual disease in T-cell acute lymphoblastic leukemia using polymerase chain reaction predicts impending relapse. 总被引:4,自引:1,他引:4
G A Neale J Menarguez G R Kitchingman T J Fitzgerald M Koehler J Mirro R M Goorha 《Blood》1991,78(3):739-747
After achieving remission, approximately one-third of patients with T-cell acute lymphoblastic leukemia (T-ALL) relapse due to the resurgence of residual leukemic cells that cannot be detected in remission by morphologic methods. Thus, the early detection of residual disease is highly desirable to monitor the efficacy of therapy, or to institute an alternative mode of therapy. Toward this aim, we have examined the applicability of polymerase chain reaction (PCR) amplification in the detection of minimal residual disease (MRD) in bone marrow samples from patients with T-ALL in morphologic remission. Two different approaches were taken to identify leukemic clone-specific sequences that could be used as targets for PCR amplification. The first technique used T-cell receptor-delta (TCR-delta) gene rearrangements that were sequenced directly after PCR amplification of leukemic DNA. This method was successful in generating clone-specific probes for 76% of T-ALL patients screened. An alternative method was used to clone and sequence a TCR-beta chain gene from leukemic cells to generate a specific probe. The PCR assays that we used were specific for each patient's leukemic clone, and were capable of routinely detecting one leukemic cell in 10(4) normal cells. Using these sensitive PCR-based assays, we found no evidence for persistence of the leukemic clone in any of the bone marrow samples from four T-ALL patients who are in long-term (3.9 + to 8.1 + years) remission. In contrast, we detected residual disease in clinical remission samples from two patients who subsequently relapsed. In one patient, where we had appropriate samples, we observed a dramatic expansion of the leukemic clone 3 months before clinical relapse. These results suggest that PCR-based assays for detection of MRD in T-ALL patients have great potential in predicting impending relapse, and in determining the efficacy of the anti-leukemic therapy. These methods may also allow the identification of long-term survivors. 相似文献
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Giuseppe Gaipa Giovanni Cazzaniga Maria Grazia Valsecchi Renate Panzer-Gr��mayer Barbara Buldini Daniela Silvestri Leonid Karawajew Oscar Maglia Richard Ratei Alessandra Benetello Simona Sala Angela Schumich Andre Schrauder Tiziana Villa Marinella Veltroni Wolf-Dieter Ludwig Valentino Conter Martin Schrappe Andrea Biondi Michael N. Dworzak Giuseppe Basso 《Haematologica》2012,97(10):1582-1593
Background
Flow cytometric analysis of leukemia-associated immunophenotypes and polymerase chain reaction-based amplification of antigen-receptor genes rearrangements are reliable methods for monitoring minimal residual disease. The aim of this study was to compare the performances of these two methodologies in the detection of minimal residual disease in childhood acute lymphoblastic leukemia.Design and Methods
Polymerase chain reaction and flow cytometry were simultaneously applied for prospective minimal residual disease measurements at days 15, 33 and 78 of induction therapy on 3565 samples from 1547 children with acute lymphoblastic leukemia enrolled into the AIEOP-BFM ALL 2000 trial.Results
The overall concordance was 80%, but different results were observed according to the time point. Most discordances were found at day 33 (concordance rate 70%) in samples that had significantly lower minimal residual disease. However, the discordance was not due to different starting materials (total versus mononucleated cells), but rather to cell input number. At day 33, cases with minimal residual disease below or above the 0.01% cut-off by both methods showed a very good outcome (5-year event-free survival, 91.6%) or a poor one (5-year event-free survival, 50.9%), respectively, whereas discordant cases showed similar event-free survival rates (around 80%).Conclusions
Within the current BFM-based protocols, flow cytometry and polymerase chain reaction cannot simply substitute each other at single time points, and the concordance rates between their results depend largely on the time at which they are used. Our findings suggest a potential complementary role of the two technologies in optimizing risk stratification in future clinical trials.Key words: childhood ALL, minimal residual disease, prognostic value 相似文献10.
Grimwade D 《Best Practice & Research: Clinical Haematology》2002,15(1):137-158
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Kwan E Norris MD Zhu L Ferrara D Marshall GM Haber M 《British journal of haematology》2000,109(2):430-434
A number of prospective studies have indicated the clinical utility of measuring minimal residual disease (MRD) in childhood acute lymphoblastic leukaemia (ALL) and have highlighted the need for improved methodology for quantification of residual disease. We describe a novel real-time polymerase chain reaction (PCR) strategy for MRD analysis based on the exonuclease activity of Taq polymerase to cleave a fluorescently labelled probe. Using a consensus probe designed to the framework 2 region of the IgH gene, together with leukaemia-specific primers, the utility of this technique for simultaneous detection and quantification of MRD was demonstrated in samples from six ALL patients. This technique provides a rapid quantitative assay for determining MRD levels which lends itself to the routine monitoring of minimal residual leukaemia. 相似文献
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Aurélie Caye Khe?ra Beldjord Kelly Mass-Malo Séverine Drunat Jean Soulier Virginie Gandemer André Baruchel Yves Bertrand Hélène Cavé Emmanuelle Clappier 《Haematologica》2013,98(4):597-601
Deletion of the Ikaros (IKZF1) gene is an oncogenic lesion frequently associated with BCR-ABL1-positive acute lymphoblastic leukemias. It is also found in a fraction of BCR-ABL1-negative B-cell precursor acute lymphoblastic leukemias, and early studies showed it was associated with a higher risk of relapse. Therefore, screening tools are needed for evaluation in treatment protocols and possible inclusion in risk stratification. Besides monosomy 7 and large 7p abnormalities encompassing IKZF1, most IKZF1 alterations are short, intragenic deletions. Based on cohorts of patients, we mapped the microdeletion breakpoints and developed a breakpoint-specific fluorescent multiplex polymerase chain reaction that allows detection of recurrent intragenic deletions. This sensitive test could also detect IKZF1 subclonal deletions, whose prognostic significance should be evaluated. Moreover, we show that consensus breakpoint sequences can be used as clonal markers to monitor minimal residual disease. This paper could be useful for translational studies and in clinical management of BCP-ALL. 相似文献
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Thermostable DNA polymerase chain amplification of t(14;18) chromosome breakpoints and detection of minimal residual disease. 总被引:25,自引:3,他引:25
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M Crescenzi M Seto G P Herzig P D Weiss R C Griffith S J Korsmeyer 《Proceedings of the National Academy of Sciences of the United States of America》1988,85(13):4869-4873
Achieving the capacity to detect minimal numbers of neoplastic cells is a major cancer diagnostic challenge. Chromosomal translocations such as the t(14;18)(q32;q21) found in follicular and some nonfollicular lymphomas provide a tumor-specific molecular marker. The 14;18 breakpoints are focused at one of six immunoglobulin heavy chain joining (JH) regions on chromosome 14 and a small major breakpoint region (MBR) of the BCL2 gene on chromosome 18. We utilized universal oligonucleotide primers of a region 5' to the BCL2 MBR and at the 3' end of JH segments to initiate a DNA polymerase chain reaction that amplified these BCL2-JH junctures. Use of thermostable DNA polymerase enabled annealing and synthesis steps at temperatures approaching the melting point of the primers, providing a sensitive and specific assay capable of detecting 1 lymphoma cell in 10(6) normal cells. This technique identified the subclinical presence of leukemic cells in all seven patients examined, including two in clinical remission. It also assessed the effectiveness of protocols designed to purge malignant cells from marrow. Moreover, this approach enabled the rapid DNA sequencing of chromosomal breakpoints without their molecular cloning. This assay markedly refines the capacity to detect minimal residual disease and should improve the ability to determine the stage of disease, stratify treatment, and evaluate therapy. 相似文献
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Use of polymerase chain reactions to monitor minimal residual disease in acute lymphoblastic leukemia patients. 总被引:8,自引:0,他引:8
S Yokota T E Hansen-Hagge W D Ludwig A Reiter A Raghavachar E Kleihauer C R Bartram 《Blood》1991,77(2):331-339
T-cell receptor (TCR) delta gene rearrangements are observed in more than 80% of acute lymphoblastic leukemia (ALL) patients. Moreover, a preferential usage of specific genetic elements has been shown in different ALL subtypes: V delta 1 DJ delta 1 rearrangements predominate in T-ALL, while most B-precursor ALLs show a recombination of V delta 2 to D delta 3. Recently we have proposed a strategy for the detection of minimal residual disease (MRD) based on the isolation of clonospecific probes following the in vitro amplification of V delta 1 DJ delta 1 junctions by polymerase chain reaction (PCR) and now have adapted this method to the preparation of specific V delta 2 D delta 3 fragments. In the present study, clonospecific probes were generated from 11 T-ALL and 16 cALL patients (21 children, 6 adults). The sensitivity of these 27 probes in detecting residual leukemia cells varied between 10(-4) to 10(-6) as determined by semiquantitative evaluation of dilution experiments. PCR analysis of 55 bone marrow (BM) and peripheral blood (PB) samples obtained from the 27 patients during complete clinical remission showed the following results: (1) Evidence for MRD was obtained in the BM of all patients (eight of eight) investigated 2 to 6 months after remission induction and also in 6 of 11 cases on maintenance therapy 7 to 19 months after diagnosis. (2) In contrast, all patients but one (10 of 11) analyzed 6 to 41 months after the termination of treatment lacked apparent evidence for leukemia DNA; the exception was a girl exhibiting 10(-4) to 10(-5) residual cells in her PB 5.5 years after diagnosis. (3) Longitudinal analysis in nine patients disclosed marked individual differences in the intervals between achievement of clinical remission and complete eradication of the leukemia cell clone. (4) Differences in the duration of MRD were not associated with distinct clinical-hematologic features. (5) Detection of residual disease by PCR proceeded clinical relapse in two cases. 相似文献
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Robillard N Cavé H Méchinaud F Guidal C Garnache-Ottou F Rohrlich PS Avet-Loiseau H Garand R 《Haematologica》2005,90(11):1516-1523
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K Miyamura M Tanimoto Y Morishima K Horibe K Yamamoto M Akatsuka Y Kodera S Kojima K Matsuyama N Hirabayashi 《Blood》1992,79(5):1366-1370
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Muñoz L López O Martino R Brunet S Bellido M Rubiol E Sierra J Nomdedéu JF 《Haematologica》2000,85(7):704-710