Skip to main content
SpringerLink
Log in

Detection of the BCR-ABL Gene by Interphase Fluorescence In Situ Hybridization (iFISH) in Chronic Myelogenous Leukemia Patients after Hemopoietic Stem Cell Transplantation: The Feasibility of iFISH Monitoring of Therapeutic Response in Peripheral Blood

  • Case Report
  • Published:
International Journal of Hematology Aims and scope Submit manuscript

Abstract

The detection of the Philadelphia (Ph) translocation has been accomplished primarily by cytogenetic analysis and reverse transcriptase polymerase chain reaction (RT-PCR). RT-PCR is highly sensitive (1/104-106) but not quantitatively reliable and is thus unsuitable for the monitoring of Ph-positive cells during therapy. Interphase fluorescence in situ hybridization (iFISH) allows analysis of a large number of cells (>500) in a timely and efficiently quantitative manner. We obtained 118 peripheral blood (PB) and 127 bone marrow (BM) samples from 75 adult chronic myelogenous leukemia (CML) patients undergoing stem cell transplantation. We simultaneously performed nested RT-PCR and iFISH for all samples. False-positive cells were detected in 2.48% ± 0.93% (mean ± SD) of PB samples and 2.75% ± 0.83% of BM samples. The iFISH results for PB and BM ranged from 1.4% to 92.8% and 1.0% to 93.8%, respectively. Correlation analysis of iFISH results for PB versus BM samples showed a strong relation (r = .993). A significant correlation (P < .05) was also found between iFISH and first-round RT-PCR. The sensitivity of BCR-ABL iFISH was similar to that of first-round RT-PCR, and iFISH results for PB and BM were also well correlated. Thus, iFISH analysis of PB and/or BM samples may be more clinically reliable than RT-PCR in the quantitative monitoring of BCR-ABL fusion in CML after transplantation.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. Nowell PC, Hungerford DA. A minute chromosome in human chronic myelocytic leukemia.Science. 1960;132:1497–1501.

    Google Scholar 

  2. Rowley JD. A new consistent chromosomal abnormality in chronic myelogenous leukemia identified by quinacrine fluorescence and Giemsa staining.Nature. 1973;243:290–293.

    Article  PubMed  CAS  Google Scholar 

  3. Groffen J, Stephenson JR, Heisterkamp N, de Klein A, Bartram CR, Grosveld G. Philadelphia chromosomal breakpoints are clustered within a limited region, bcr, on chromosome 22.Cell. 1984;36:93–99.

    Article  PubMed  CAS  Google Scholar 

  4. Shtivelman E, Lifshitz B, Gale RP, Canaani E. Fused transcript of abl and bcr genes in chronic myelogenous leukaemia.Nature. 1985;315:550–554.

    Article  PubMed  CAS  Google Scholar 

  5. Arnoldus EP, Wiegant J, Noordermeer IA, et al. Detection of the Philadelphia chromosome in interphase nuclei.Cytogenet Cell Genet. 1990;54:108–111.

    Article  PubMed  CAS  Google Scholar 

  6. Tkachuk DC, Westbrook A, Andreeff M, et al. Detection of bcr-abl fusion in chronic myelogenous leukemia by in situ hybridization.Science. 1990;250:559–562.

    Article  PubMed  CAS  Google Scholar 

  7. Dewald GW, Schad CR, Christensen ER, et al. The application of fluorescent in situ hybridization to detect Mbcr/abl fusion in variants Ph chromosomes in CML and ALL.Cancer Genet Cytogenet. 1993;71:7–14.

    Article  PubMed  CAS  Google Scholar 

  8. Bentz M, Cabot G, Moos M, et al. Detection of chimeric BCR-ABL genes on bone marrow samples and blood smears in chronic myeloid and acute lymphoblastic leukemia by in situ hybridization.Blood. 1994;83:1922–1928.

    PubMed  CAS  Google Scholar 

  9. Froncillo MC, Maffei L, Cantonetti M, et al. FISH analysis for CML monitoring?Ann Hematol. 1996;73:113–119.

    Article  PubMed  CAS  Google Scholar 

  10. Morgan GJ, Hughes T, Janssen JW, et al. Polymerase chain reaction for detection of residual leukemia.Lancet. 1989;1:928–929.

    Article  PubMed  CAS  Google Scholar 

  11. Sklar J. Polymerase chain reaction: the molecular microscope of residual disease.J Clin Oncol. 1991;9:1521–1524.

    Article  PubMed  CAS  Google Scholar 

  12. Chomczynski P, Sacchi N. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction.Anal Biochem. 1987;162:156–159.

    Article  PubMed  CAS  Google Scholar 

  13. Kwok S, Higuchi R. Avoiding false positives with PCR.Nature. 1989;339:237–238.

    Article  PubMed  CAS  Google Scholar 

  14. Bland JM, Altman DG. Statistical methods for assessing agreement between two methods of clinical measurement.Lancet. 1986;1:307–310.

    Article  PubMed  CAS  Google Scholar 

  15. Lukasova E, Kozubek S, Kozubek M, et al. Localisation and distance between ABL and BCR genes in interphase nuclei of bone marrow cells of control donors and patients with chronic myeloid leukaemia.Hum Genet. 1997;100:525–535.

    Article  PubMed  CAS  Google Scholar 

  16. Clarkson B, Strife A. Linkage of proliferative and maturational abnormalities in chronic myelogenous leukemia and relevance to treatment.Leukemia. 1993;7:1683–1721.

    PubMed  CAS  Google Scholar 

  17. Morley A. Quantifying leukemia.N Engl J Med. 1998;339:627–629.

    Article  PubMed  CAS  Google Scholar 

  18. Biernaux C, Loos M, Sels A, Huez G, Stryckmans P. Detection of major bcr-abl gene expression at a very low level in blood cells of some healthy individuals.Blood. 1995;86:3118–3122.

    PubMed  CAS  Google Scholar 

  19. Bose S, Deininger M, Gora-Tybor J, Goldman JM, Melo JV. The presence of typical and atypical BCR-ABL fusion genes in leukocytes of normal individuals: biologic significance and implications for the assessment of minimal residual disease.Blood. 1998;92:3362–3367.

    PubMed  CAS  Google Scholar 

  20. Faderl S, Talpaz M, Kantarjian HM, Estrov Z. Should polymerase chain reaction analysis to detect minimal residual disease in patients with chronic myelogenous leukemia be used in clinical decision making?Blood. 1999;93:2755–2759.

    PubMed  CAS  Google Scholar 

  21. Garcia-Isidoro M, Tabernero MD, Garcia JL, et al. Detection of the Mbcr/abl translocation in chronic myeloid leukemia by fluorescence in situ hybridization: comparison with conventional cytogenetics and implications for minimal residual disease detection.Hum Pathol. 1997;28:154–159.

    Article  PubMed  CAS  Google Scholar 

  22. Muhlmann J, Thaler J, Hilbe W, et al. Fluorescence in situ hybridization (FISH) on peripheral blood smears for monitoring Philadelphia chromosome-positive chronic myeloid leukemia (CML) during interferon treatment: a new strategy for remission assessment.Genes Chromosomes Cancer. 1998;21:90–100.

    Article  PubMed  CAS  Google Scholar 

  23. Yanagi M, Shinjo K, Takeshita A, et al. Simple and reliably sensitive diagnosis and monitoring of Philadelphia chromosome-positive cells in chronic myeloid leukemia by interphase fluorescence in situ hybridization of peripheral blood cells.Leukemia. 1999;13:542–552.

    Article  PubMed  CAS  Google Scholar 

  24. Dewald GW, Wyatt WA, Juneau AL, et al. Highly sensitive fluorescence in situ hybridization method to detect double BCR/ABL fusion and monitor response to therapy in chronic myeloid leukemia.Blood. 1998;91:3357–3365.

    PubMed  CAS  Google Scholar 

  25. Sinclair PB, Green AR, Grace C, Nacheva EP. Improved sensitivity of BCR-ABL detection: a triple-probe three-color fluorescence in situ hybridizaton system.Blood. 1997;90:1395–1402.

    PubMed  CAS  Google Scholar 

  26. Buno I, Wyatt WA, Zinsmeister AR, Dietz-Band J, Silver RT, Dewald GW. A special fluorescent in situ hybridization technique to study peripheral blood and assess the effectiveness of interferon therapy in chronic myeloid leukemia.Blood. 1998;92:2315–2321.

    PubMed  CAS  Google Scholar 

  27. Dewald GW, Wyatt WA, Silver RT. Atypical BCR and ABL D-FISH patterns in chronic myeloid leukemia and their possible role in therapy.Leuk Lymphoma. 1999;34:481–491.

    Article  PubMed  CAS  Google Scholar 

  28. Mohr B, Bornhauser M, Platzbecker U, et al. Problems with interphase fluorescence in situ hybridization in detecting BCR/ABL-positive cells in some patients using a novel technique with extra signals.Cancer Genet Cytogenet. 2001;27:111–117.

    Article  Google Scholar 

  29. Mensink E, van de Locht A, Schattenberg A, et al. Quantitation of minimal residual disease in Philadelphia chromosome positive chronic myeloid leukaemia patients using real-time quantitative RT-PCR.Br J Haematol. 1998;102:768–774.

    Article  PubMed  CAS  Google Scholar 

  30. Eder M, Battmer K, Kafert S, Stucki A, Ganser A, Hertenstein B. Monitoring of BCR-ABL expression using real-time RT-PCR in CML after bone marrow or peripheral blood stem cell transplantation.Leukemia. 1999;13:1383–1389.

    Article  PubMed  CAS  Google Scholar 

  31. Chomel JC, Brizard F, Veinstein A, et al. Persistence of BCR-ABL genomic rearrangement in chronic myeloid leukemia patients in complete and sustained cytogenetic remission after interferonalpha therapy or allogeneic bone marrow transplantation.Blood. 2000;95:404–408.

    PubMed  CAS  Google Scholar 

  32. Le Gouill S, Talmant P, Milpied N, et al. Fluorescence in situ hybridization on peripheral-blood specimens is a reliable method to evaluate cytogenetic response in chronic myeloid leukemia.J Clin Oncol. 2000;18:1533–1538.

    Article  PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Clinical Pathology, College of Medicine, Soonchunhyang University, Seoul, Korea

    You Kyoung Lee & Dong Wha Lee

  2. Department of Catholic Hemopoietic Stem Cell Transplantation Center, St. Mary’s Hospital, The Catholic University of Korea, #62 Youido-dong Yeungdeungpo-gu, 150-713, Seoul, Korea

    Yoo Li Kim, Seok Lee, Chang Ki Min, Yoo-Jin Kim, Il-Hoan Oh, Tai-Gyu Kim, Chun Choo Kim & Dong-Wook Kim

Authors
  1. You Kyoung Lee
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Dong Wha Lee
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Yoo Li Kim
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Seok Lee
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Chang Ki Min
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Yoo-Jin Kim
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Il-Hoan Oh
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Tai-Gyu Kim
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Chun Choo Kim
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Dong-Wook Kim
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Dong-Wook Kim.

About this article

Cite this article

Lee, Y.K., Lee, D.W., Kim, Y.L. et al. Detection of the BCR-ABL Gene by Interphase Fluorescence In Situ Hybridization (iFISH) in Chronic Myelogenous Leukemia Patients after Hemopoietic Stem Cell Transplantation: The Feasibility of iFISH Monitoring of Therapeutic Response in Peripheral Blood. Int J Hematol 76, 180–185 (2002). https://doi.org/10.1007/BF02982582

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF02982582

Key words

Search

Navigation