Detection and quantitation of neoplastic cells in acute lymphoblastic leukaemia, by use of the polymerase chain reaction

Brisco, Michael J.; Condon, John T.; Sykes, Pamela J.; Neoh, S. H.; Morley, Alexander A.

doi:10.1111/j.1365-2141.1991.tb04524.x

Cited by 68 publications

(23 citation statements)

References 22 publications

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“…Each result was then converted into estimates of the actual amounts of amplifiable gDNA in mg and multiplied by 3.3 Â 10 5 to estimate the number of amplifiable haploid genomes. 22,23 PCRs were performed using PrimerDesign (Southampton, UK) master mix under standard conditions, with 5 ml of gDNA (approximately 100 ng). PCRs were set up on a RotorGene CAS1200 robot and run on the RotorGene 6000 platform (Cambridge, UK).…”

Section: Absolute Quantification Of Gdna Fusions In Pre-imatinib Samplesmentioning

confidence: 99%

Detection and molecular monitoring of FIP1L1-PDGFRA-positive disease by analysis of patient-specific genomic DNA fusion junctions

et al. 2008

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To evaluate current detection methods for FIP1L1-PDGFRA in hypereosinophilic syndrome (HES), we developed a means to rapidly amplify genomic break points. We screened 202 cases and detected genomic junctions in all samples previously identified as RT-PCR positive (n ¼ 43). Genomic fusions were amplified by single step PCR in all cases whereas only 22 (51%) were single step RT-PCR positive. Importantly, FIP1L1-PDGFRA was detected in two cases that initially tested negative by RT-PCR or fluorescence in situ hybridization. Absolute quantitation of the fusion by real-time PCR from genomic DNA (gDNA) using patient-specific primer/probe combinations at presentation (n ¼ 13) revealed a 40-fold variation between patients (range, 0.027-1.1 FIP1L1-PDGFRA copies/haploid genome). In follow up samples, quantitative analysis of gDNA gave 1-2 log greater sensitivity than RQ-PCR of cDNA. Minimal residual disease assessment using gDNA showed that 11 of 13 patients achieved complete molecular response to imatinib within a median of 9 months (range, 3-17) of starting treatment, with a sensitivity of detection of up to 1 in 10 5 . One case relapsed with an acquired D842V mutation. We conclude that detection of FIP1L1-PDGFRA from gDNA is a useful adjunct to standard diagnostic procedures and enables more sensitive follow up of positive cases after treatment.

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Section: Absolute Quantification Of Gdna Fusions In Pre-imatinib Samplesmentioning

confidence: 99%

Detection and molecular monitoring of FIP1L1-PDGFRA-positive disease by analysis of patient-specific genomic DNA fusion junctions

et al. 2008

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“…17 For each patient the rearranged IgH gene was sequenced and a pair of 'leukemia-specific' primers was synthesized. The numbers of amplifiable leukemic targets and amplifiable N-ras targets were quantified by PCR using limiting dilution analysis, which involved multiple replicates and Poisson statistics, 12 and the proportion of nucleated marrow cells which belong to the leukemic clone was calculated assuming each leukemic cell contains one rearranged IgH gene and all cells contain two N-ras genes.…”

Section: Sensitive Quantification Of Mrdmentioning

confidence: 99%

Comparison of methods for assessment of minimal residual disease in childhood B-lineage acute lymphoblastic leukemia

et al. 2001

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The level of minimal residual disease (MRD) early in treatment of acute lymphoblastic leukemia (ALL) strongly predicts the risk of marrow relapse. As a variety of methods of varying complexity have been separately used for detecting and quantifying MRD, we compared the prognostic utility of three methodsmeasurement of blast percentage on day 14 of treatment, detection of monoclonality on day 14 or day 35, and measurement of MRD by PCR-based limiting dilution analysis on day 14 or day 35. The study group comprised 38 children aged 1-15 with Philadelphia-negative B-lineage ALL who were uniformly treated and followed until relapse or for a minimum of 5 years. We also studied some of the technical factors which influence the ability to detect MRD. Measurement of blast percentage on day 14 by an expert morphologist, detection of monoclonality on day 35, and PCR-based measurement of MRD levels on days 14 and 35 all showed significant ability to divide patients into prognostic groups. Measurement of blast percentage on day 14 by routine morphology or detection of monoclonality on day 14 were not useful. The quality of DNA samples varied greatly, as determined by amplifiability in the PCR. However, virtually all amplifiable leukemic targets in a sample were detectable which suggests that the level of detection achieved by limiting dilution analysis is essentially determined by the amount of DNA which it is practicable to study. We conclude that quantification of MRD at the end of induction provides the full range of prognostic information for marrow relapse but is complex; detection of monoclonality on day 35 is simple and has good positive predictive value; and quantification of MRD on day 14 merits further study. PCR-based methods for measurement of MRD levels should incorporate a correction for variation in DNA amplifiability. Leukemia (2001) 15, 385-390.

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“…[1][2][3][4][5][6][7][8][9][10][11][12][13][14][15][16] MRD may be detected at a sensitivity of at least 10 À4 either by molecular means, generally based on polymerase chain reaction (PCR) amplification of rearranged immunoglobulin (Ig) or T-cell receptor genes 13,14,[16][17][18][19][20] or by multiparameter flow cytometry. 1,2,[8][9][10]15,[21][22][23][24][25][26] The latter approach is based on the fact that leukemic cells express aberrant phenotypic combinations of antigens that are not present on normal bone marrow cells. 9,22,[27][28][29] Both methods are generally recognized as useful, although they have different advantages and disadvantages.…”

Section: Introductionmentioning

confidence: 99%

Minimal residual disease detection in childhood precursor–B-cell acute lymphoblastic leukemia: relation to other risk factors. A Children's Oncology Group study

et al. 2003

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Minimal residual disease (MRD) can be detected in the marrows of children undergoing chemotherapy either by flow cytometry or polymerase chain reaction. In this study, we used four-color flow cytometry to detect MRD in 1016 children undergoing therapy on Children's Oncology Group therapeutic protocols for precursor-B-cell ALL. Compliance was excellent, with follow-up samples received at the end of induction on nearly 95% of cases; sensitivity of detection at this time point was at least 1/10,000 in more than 90% of cases. Overall, 28.6% of patients had detectable MRD at the end of induction. Patients with M3 marrows at day 8 were much more likely to be MRD positive (MRD+) than those with M2 or M1 marrows. Different genetically defined groups of patients varied in their prevalence of MRD. Specifically, almost all patients with BCR-ABL had high levels of end-of-induction MRD. Only 8.4% of patients with TEL-AML1 were MRD+40.01% compared with 20.3% of patients with trisomies of chromosomes 4 and 10. Our results show that MRD correlates with conventional measures of slow early response. However, the high frequency of MRD positivity in favorable trisomy patients suggests that the clinical significance of MRD positivity at the end of induction may not be the same in all patient groups.

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Detection and quantitation of neoplastic cells in acute lymphoblastic leukaemia, by use of the polymerase chain reaction

Cited by 68 publications

References 22 publications

Detection and molecular monitoring of FIP1L1-PDGFRA-positive disease by analysis of patient-specific genomic DNA fusion junctions

Detection and molecular monitoring of FIP1L1-PDGFRA-positive disease by analysis of patient-specific genomic DNA fusion junctions

Comparison of methods for assessment of minimal residual disease in childhood B-lineage acute lymphoblastic leukemia

Minimal residual disease detection in childhood precursor–B-cell acute lymphoblastic leukemia: relation to other risk factors. A Children's Oncology Group study

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