Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Original Manuscript
  • Published:

Chronic Myeloid Leukemia (CML)

Comparison of chromosome banding analysis, interphase- and hypermetaphase-FISH, qualitative and quantitative PCR for diagnosis and for follow-up in chronic myeloid leukemia: a study on 350 cases

Leukemia volume 16, pages 53–59 (2002)Cite this article

Abstract

For the diagnosis of CML and for monitoring of treatment response the detection of the t(9;22)(q34;q11) or the BCR-ABL rearrangement is necessary. Chromosome banding analysis (CA) is still the gold standard but other techniques like Southern blot, fluorescence in situ hybridization (FISH) and polymerase chain reaction (PCR) are available. We analyzed 350 CML patients at different stages of disease in parallel with CA, interphase-FISH (IP-FISH), hypermetaphase-FISH (HM-FISH) and RT-PCR. In 20 cases with no Ph+ metaphases in CA, HM-FISH detected 0.2 to 10% BCR-ABL+metaphases. After IP-FISH 107 samples were judged as negative. However, in 17 of these samples HM-FISH detected BCR-ABL+ metaphases (0.3–11%), and in eight cases CA detected Ph+ metaphases (2.5–25%). A comparison of IP-FISH performed on uncultivated cells vs cells cultivated for 48 h in 70 cases revealed a higher proportion of BCR-ABL+ cells in the cultivated samples. If nested PCR was negative, all other methods were negative in all cases too. In addition, 94 cases were evaluated using real-time PCR (LightCycler technology). The BCR-ABL/cABL ratio measured showed a high correlation with all other methods. Interestingly, a wide range in the BCR-ABL/ABL ratio was observed especially in patients who showed 100% Ph-positive metaphases in CA. In conclusion, CA, IP-FISH, HM-FISH and real-time PCR give reliable results but differences due to measurement of different target structures have to be kept in mind when using these data for definition of remission status.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1
Figure 2
Figure 3

Similar content being viewed by others

References

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

    Article  CAS  Google Scholar 

  2. Davis RL, Konopka JB, Witte ON . Activation of the c-abl oncogene by viral transduction generates altered c-abl proteins with similar in vitro kinase properties Mol Cell Biol 1985 5: 204–213

    Article  CAS  Google Scholar 

  3. Daley GQ, Van Etten RA, Baltimore D . Induction of chronic myelogenous leukemia in mice by the P210bcr/abl gene of the Philadelphia chromosome Science 1990 247: 824–830

    Article  CAS  Google Scholar 

  4. Ganesan TS, Rassool R, Guo A-P, Th'ng KH, Dowding C, Hibbin JA, Young BD, White H, Kumaran TO, Galton DAG, Goldman JM . Rearrangement of the bcr gene in Philadelphia chromosome-negative chronic myeloid leukemia Blood 1986 68: 957–960

    CAS  PubMed  Google Scholar 

  5. Tkachuk DC, Westbrook CA, Andreeff M, Donlon TA, Cleary ML, Suryanarayan K, Homge M, Redner A, Gray J, Pinkel D . Detection of bcr-abl fusion in chronic myelogeneous leukemia by in situ hybridization Science 1990 250: 559–562

    Article  CAS  Google Scholar 

  6. Kawasaki ES, Clark SS, Coyne MY, Smith SD, Champlin R, Witte ON, McCormick FP . Diagnosis of chronic myeloid and acute lymphocytic leukemias by detection of leukemia-specific mRNA sequences amplified in vitro Proc Natl Acad Sci USA 1988 85: 5698–5702

    Article  CAS  Google Scholar 

  7. Reiter A, Skladny H, Hochhaus A, Seifarth W, Heimpel H, Bartram CR, Cross NCP, Hehlmann R . Molecular response of CML patients treated with interferon-a monitored by quantitative Southern blot analysis Br J Haematol 1997 97: 86–93

    Article  CAS  Google Scholar 

  8. Hochhaus A, Weisser A, La Rosée P, Emig M, Müller MC, Saussele S, Reiter A, Kuhn C, Berger U, Hehlmann R, Cross NCP . Detection and quantification of residual disease in chronic myelogenous leukemia Leukemia 2000 14: 998–1005

    Article  CAS  Google Scholar 

  9. Nanjangud G, Kadam PR, Saikia T, Bhisey AN, Kumar A, Gopal R, Chopra H, Nair CN, Advani SH . Karyotypic findings as an independent prognostic marker in chronic myeloid leukaemia blast crisis Leuk Res 1994 18: 385–392

    Article  CAS  Google Scholar 

  10. Mitelman F . The cytogenetic scenario of chronic myeloid leukemia Leuk Lymphoma 1993 11: 11–15

    Article  Google Scholar 

  11. Cross NC . Minimal residual disease in chronic myeloid leukaemia Hematol Cell Ther 1998 40: 224–228

    CAS  PubMed  Google Scholar 

  12. Seong DC, Kantarjian HM, Ro JY, Talpaz M, Xu J, Robinson JR, Deisseroth AB, Champlin RE, Siciliano MJ . Hypermetaphase fluorescence in situ hybridization for quantitative monitoring of Philadelphia chromosome-positive cells in patients with chronic myelogenous leukemia during treatment Blood 1995 86: 2343–2349

    CAS  PubMed  Google Scholar 

  13. Lion T, Izraeli S, Henn T, Gaiger A, Mor W, Gadner H . Monitoring of residual disease in chronic myelogenous leukemia by quantitative polymerase chain reaction Leukemia 1992 6: 495–499

    CAS  PubMed  Google Scholar 

  14. Emig M, Saussele S, Wittor H, Weisser A, Reiter A, Willer A, Berger U, Hehlmann R, Cross NC, Hochhaus A . Accurate and rapid analysis of residual disease in patients with CML using specific fluorescent hybridization probes for real time quantitative RT-PCR Leukemia 1999 13: 1825–1832

    Article  CAS  Google Scholar 

  15. Preudhomme C, Revillion F, Merlat A, Hornez L, Roumier C, Duflos Grardel N, Jouet JP, Cosson A, Peyrat JP, Fenaux P . Detection of BCR-ABL transcripts in chronic myeloid leukemia (CML) using a ‘real time’ quantitative RT-PCR assay Leukemia 1999 13: 957–964

    Article  CAS  Google Scholar 

  16. Fonatsch C, Schaadt M, Kirchner H, Diehl V . A possible correlation between the degree of karyotype aberrations and the rate of sister chromatid exchanges in lymphoma lines Int J Cancer 1980 26: 749–756

    Article  CAS  Google Scholar 

  17. ISCN 1995 In: Mitelman F (ed.). Guidelines for Cancer Cytogenetics, Supplement to: An International System for HumanCytogenetic Nomenclature S Karger: Basel 1995

  18. Kantarjian HM, Smith TL, O'Brien S, Beran M, Pierce S, Talpaz M . Prolonged survival in chronic myelogenous leukemia after cytogenetic response to interferon-alpha therapy. The Leukemia Service Ann Intern Med 1995 122: 254–261

    Article  CAS  Google Scholar 

  19. Maurer J, Janssen JWG, Thiel E, van Denderen J, Ludwig W-D, Aydemir Ü, Heinze B, Fonatsch C, Harbott J, Reiter A, Riehm H, Hoelzer D, Bartram CR . Detection of chimeric BCR-ABL genes in acute lymphoblastic leukaemia by polymerase chain reaction Lancet 1991 337: 1055–1058

    Article  CAS  Google Scholar 

  20. Seong CM, Giralt S, Kantarjian H, Xu J, Swantkowski J, Hayes K, Glassman AB, Khouri I, Korbling M, Thall P, Siciliano MJ, Champlin RE . Early detection of relapse by hypermetaphase fluorescence in situ hybridization after allogeneic bone marrow transplantation for chronic myeloid leukemia J Clin Oncol 2000 18: 1831–1836

    Article  CAS  Google Scholar 

  21. Wilkens L, Varga S, Werner M, Nolte M, v Wasielewski R, Georgii A . Short-term culturing influences the number of bcr/abl-fused cells detected by fluorescence in situ hybridisation in bone marrow aspirates of CML patients Pathobiology 1999 67: 163–168

    Article  CAS  Google Scholar 

  22. Cuneo A, Bigoni R, Emmanuel B, Smit E, Rigolin GM, Roberti MG, Bardi A, Piva N, Scapoli G, Castoldi G, Van Den Berghe H, Hagemeijer A . Fluorescence in situ hybridization for the detection and monitoring of the Ph-positive clone in chronic myelogenous leukemia: comparison with metaphase banding analysis Leukemia 1998 12: 1718–1723

    Article  CAS  Google Scholar 

  23. 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

    CAS  PubMed  Google Scholar 

  24. Muhlmann J, Thaler J, Hilbe W, Bechter O, Erdel M, Utermann G, Duba HC . 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  CAS  Google Scholar 

  25. Le Gouill S, Talmant P, Milpied N, Daviet A, Ancelot M, Moreau P, Harousseau JL, Bataille R, Avet Loiseau H . 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  CAS  Google Scholar 

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

    CAS  PubMed  Google Scholar 

  27. Dewald GW, Wyatt WA, Juneau AL, Carlson RO, Zinsmeister AR, Jalal SM, Spurbeck JL, Silver RT . 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

    CAS  PubMed  Google Scholar 

  28. Radich JP, Gehly G, Gooley T, Bryant E, Clift RA, Collins S, Edmands S, Kirk J, Lee A, Kessler P . Polymerase chain-reaction detection for the bcr-abl fusion transcript after allogeneic marrow transplantation for chronic myeloid leukemia: results and implication in 346 patients Blood 1995 85: 2632–2640

    CAS  PubMed  Google Scholar 

Download references

Acknowledgements

This study was supported by grants from the German José Carreras-Foundation to TH and AH and is part of the Kompetenznetz ‘Akute und chronische Leukämien’ supported by the German Bundesminister für Bildung und Forschung.

Author information

Authors and Affiliations

  1. Department of Internal Medicine III, University Hospital Grosshadern, Ludwig-Maximilians-University, Munich, Germany

    C Schoch, S Schnittger, S Bursch, D Gerstner, W Hiddemann & T Haferlach

  2. III Medizinische Klinik, Klinikum Mannheim, Universität Heidelberg, Germany

    A Hochhaus, U Berger & R Hehlmann

  3. Clinical Cooperative Group ‘Pathogenesis of Acute Myeloid Leukemia’ of the National Center for Environment and Health (GSF), Munich, Germany

    W Hiddemann

Authors
  1. C Schoch
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. S Schnittger
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. S Bursch
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. D Gerstner
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. A Hochhaus
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. U Berger
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. R Hehlmann
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. W Hiddemann
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. T Haferlach
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Schoch, C., Schnittger, S., Bursch, S. et al. Comparison of chromosome banding analysis, interphase- and hypermetaphase-FISH, qualitative and quantitative PCR for diagnosis and for follow-up in chronic myeloid leukemia: a study on 350 cases. Leukemia 16, 53–59 (2002). https://doi.org/10.1038/sj.leu.2402329

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/sj.leu.2402329

Keywords

This article is cited by

Search

Advanced search

Quick links