Highly Sensitive Fluorescence In Situ Hybridization Method to Detect Double BCR/ABL Fusion and Monitor Response to Therapy in Chronic Myeloid Leukemia

Dewald, Gordon W.; Wyatt, William A.; Juneau, Amy L.; Carlson, Richard O.; Zinsmeister, Alan R.; Jalal, Syed M.; Spurbeck, Jack L.; Silver, Richard T.

doi:10.1182/blood.v91.9.3357

Cited by 161 publications

(38 citation statements)

References 11 publications

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“…Finally, the slides were removed from the Autostainer, rinsed in water, and counterstained in hematoxylin for 1 minute. Cytoprep slides were prepared from pooled EEC cells, and fluorescence in situ hybridization (FISH) analysis was performed with relevant probes as previously described [20].…”

Section: Immunostaining and Fluorescence In Situ Hybridization Analysismentioning

confidence: 99%

Extending Jak2V617F and MplW515 Mutation Analysis to Single Hematopoietic Colonies and B and T Lymphocytes

et al. 2007

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JAK2V617F and MPLW515L/K are myeloproliferative disorder (MPD)-associated mutations. We genotyped 552 individual hematopoietic colonies obtained by CD34؉ cell culture from 16 affected patients (13 JAK2V617F and 3 MPLW515L/K) to determine (a) the proportion of colonies harboring a particular mutation in the presence or absence of cytokines, (b) the lineage distribution of endogenous colonies for each mutation, and (c) the differences (if any) in the pattern of mutation among the various MPDs, as established by genotyping of individual colonies. Genotyping analysis revealed cohabitation of mutation-negative and mutationpositive endogenous colonies in polycythemia vera as well as other MPDs. Culture of progenitor cells harboring MPLW515L/K yielded virtually no endogenous erythroid colonies in contrast to JAK2V617F-harboring progenitor cells. The mutation pattern (i.e., relative distribution of homozygous, heterozygous, or wild-type colonies) was not a distinguishing feature among the MPDs, and MPLW515 mutations were detected in B and/or T lymphocytes in all three patients tested. These observations suggest that clonal myelopoiesis antedates acquisition of JAK2V617F or MPLW515L/K mutations and that the latter is acquired in a lympho-myeloid progenitor cell. STEM CELLS 2007;25: 2358 -2362 Disclosure of potential conflicts of interest is found at the end of this article.

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Section: Immunostaining and Fluorescence In Situ Hybridization Analysismentioning

confidence: 99%

Extending Jak2V617F and MplW515 Mutation Analysis to Single Hematopoietic Colonies and B and T Lymphocytes

et al. 2007

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“…27 The cutoff for FISH in our study was 3.5 percent (mean ± 3SD). This detection limit might be improved to less than 1 percent, as has been shown in other studies with probe sets that span both the abl and bcr breakpoints 28 or that are labeled with three different colors. 29 In our patients, no bcr/abl-negative leukapheresis components were obtained, and an increasing percentage of bcr/abl positivity was seen.…”

Section: Discussionmentioning

confidence: 90%

PBPC mobilization with chemotherapy and G–CSF in patients with chronic myeloid leukemia: quantification of bcr/abl‐positive cells by interphase fluorescence in situ hybridization and competitive PCR

et al. 2001

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Both FISH and QC-PCR were reliable methods of quantifying bcr/abl positivity, and they allowed selection of the optimal apheresis component for autologous transplantation. In both methods, a significant increase in bcr/abl positivity was seen from the first to the last leukapheresis. With FISH, results can be obtained within 24 hours. This method may prevent additional contaminated leukapheresis in case of increasing percentages of bcr/abl-positive cells.

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“…This may be related to the high difference between the number of cells analyzed by both methods and the relatively low sensitivity of FISH analysis (normal cut-off value is 8%). Dewald et al (1998) described an innovation of FISH with very high sensitivity and very low normal cut-off value (<1%) using a combination of DNA probes that detect double BCR/ABL fusion (D-FISH) on the abnormal chromosomes 9 and 22. The evaluation of PB smears by D-FISH and the correction of the results for lymphocytes or the T-cell subset might be the best alternative for BM methods of cytogenetic analysis in CML.…”

Section: Discussionmentioning

confidence: 99%

Efficiency of interphase fluorescence in situ hybridization for BCR/ABL on peripheral blood smears for monitoring of CML patients: a comparison with bone marrow findings

Akel

Kοlialexi

Mavrou

et al. 2002

Clinical &Amp; Laboratory Haematology

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Conventional cytogenetic analysis (CCA) is the standard method for monitoring of the Philadelphia (Ph) chromosome in chronic myeloid leukemia (CML). Evaluation of breakpoint cluster region/abelson murine leukemia (BCR/ABL) fusion using interphase fluorescence in situ hybridization on peripheral blood smears (PB-FISH) might be another approach allowing more frequent and less invasive follow-up investigations. Herein, BCR/ABL fusion gene was assessed on 21 PB smears from 16 CML patients in chronic phase. Results of PB-FISH were compared with those of CCA and interphase FISH on bone marrow aspirates (BM-FISH). PB-FISH analysis was combined with CD3 immunophenotyping that allowed simultaneous investigation of the leukemic status of CD3(+) T lymphocytes and scoring CD3(-) cells for BCR/ABL fusion gene. Moreover, the frequency of BCR/ABL fusion in nonlymphoid PB cells was estimated according to the differential leukocyte counts. The incidence of BCR/ABL(+) fusion signals in CD3(+) T cells of CML patients was 5.3% (SD +/- 1.9) and did not exceed the normal cut-off value of 8%. A significant correlation (P < 0.001) was found between results of PB-FISH and methods of BM analysis (CCA or BM-FISH). Correction of PB-FISH results to include only nonlymphoid or CD3(-) cells reduced the mean of differences and improved agreement between PB-FISH and CCA or BM-FISH methods. The best agreement was noted between CCA and PB-FISH on nonlymphoid cells. On the other hand, results of BM-FISH agreed well with those of PB-FISH on CD3(-) cells. These findings imply that PB-FISH on nonlymphoid or CD3(-) cells is reliable and may replace BM analysis for monitoring of response to treatment in CML patients.

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Highly Sensitive Fluorescence In Situ Hybridization Method to Detect Double BCR/ABL Fusion and Monitor Response to Therapy in Chronic Myeloid Leukemia

Cited by 161 publications

References 11 publications

Extending Jak2V617F and MplW515 Mutation Analysis to Single Hematopoietic Colonies and B and T Lymphocytes

Extending Jak2V617F and MplW515 Mutation Analysis to Single Hematopoietic Colonies and B and T Lymphocytes

PBPC mobilization with chemotherapy and G–CSF in patients with chronic myeloid leukemia: quantification of bcr/abl‐positive cells by interphase fluorescence in situ hybridization and competitive PCR

Efficiency of interphase fluorescence in situ hybridization for BCR/ABL on peripheral blood smears for monitoring of CML patients: a comparison with bone marrow findings

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