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Comparison of FISH and Quantitative RT-PCR for the Diagnosis and Follow-Up of BCR-ABL-Positive Leukemias

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Abstract

Background: For Philadelphia chromosome positive (Ph+) leukemias (chronic myelogenous leukemia [CML], acute lymphoblastic leukemia [ALL], and rare other leukemias), both allogeneic transplantation and treatment with tyrosine kinase inhibitors offer chances of molecular remission (the molecular marker being consistently undetectable). Molecular remission is defined as a reduction in the quantification of BCR-ABL transcripts to an undetectable level by molecular diagnostic methods, and is considered as a surrogate marker for cure or long-term disease control. The molecular diagnostic methods including fluorescence in situ hybridization (FISH) and quantitative reverse transcription-polymerase chain reaction (QRT-PCR) are more sensitive than classical cytogenetic analysis for the detection of BCR-ABL positive cells. QRT-PCR, due to its superior sensitivity, is considered the gold standard for the follow-up of Ph+ leukemias treated with imatinib.

Aim: The objective of our study was to compare the diagnostic and clinical usefulness of FISH and QRT-PCR at different timepoints for Ph+ leukemias.

Patients and methods: We investigated 23 unselected patients with Ph+ CML (n = 21) or Ph+ ALL (n = 2) at 77 different timepoints in a comparative study with both FISH and QRT-PCR using commercially available reagents in a routine laboratory.

Results: Our study demonstrated a good correlation of QRT-PCR with FISH in detecting the BCR-ABL fusion gene among patients with CML or ALL (coefficient of correlation = 0.77493, p < 0.0001, using Spearman’s correlation procedure). All newly diagnosed or untreated cases were positive with both methods. Lower coefficients of correlation were found when FISH and QRT-PCR were correlated with the white blood cell count (WBC). An overall concordance of FISH and QRT-PCR (being either negative or positive in both tests) was found in 65 cases (84.4%) and a discrepancy identified in 12 cases (15.6%).

Conclusions: We confirm that QRT-PCR allows precise measurement of low levels of BCR-ABL transcripts and can serve as a sensitive indicator for minimal residual disease. In addition, we demonstrate in most cases a good correlation of QRT-PCR with FISH in detecting the BCR-ABL fusion gene among patients with CML or Ph+ ALL. FISH is not suitable for monitoring minimal residual disease.

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  1. The use of trade names is for product identification purposes only and does not imply endorsement.

References

  1. Sawyers CL. Chronic myeloid leukemia. N Engl J Med 1999; 340: 1330–40

    Article  PubMed  CAS  Google Scholar 

  2. Talpaz M. Interferon-alpha-based treatment of chronic myeloid leukemia and implications of signal transduction inhibition. Semin Hematol 2001; 38(3 Suppl. 8): 22–7

    Article  PubMed  CAS  Google Scholar 

  3. Gratwohl A, Brand R, Apperley J, et al. Allogeneic hematopoietic stem cell transplantation for chronic myeloid leukemia in Europe 2006: transplant activity, long-term data and current results. An analysis by the Chronic Leukemia Working Party of the EBMT. Haematologica 2006; 91: 513–21

    PubMed  Google Scholar 

  4. Deininger M, Buchdunger E, Druker BJ. The development of imatinib as a therapeutic agent for chronic myeloid leukemia. Blood 2005; 105: 2640–53

    Article  PubMed  CAS  Google Scholar 

  5. Kebriaei P, Larson RA. Progress and challenges in the therapy of adult acute lymphoblastic leukemia. Curr Opin Hematol 2003; 10: 284–9

    Article  PubMed  Google Scholar 

  6. Goldman J. Monitoring minimal residual disease in BCR-ABL positive chronic myeloid leukemia in the imatinib era. Curr Opin Hematol 2004; 12: 33–9

    Article  Google Scholar 

  7. Martinelli G, Iacobucci I, Rosti G, et al. Prediction of response to imatinib by prospective quantitation of BCR-ABL transcript in late chronic phase chronic myeloid leukemia patients. Ann Oncol 2006; 17: 495–502

    Article  PubMed  CAS  Google Scholar 

  8. Hughes T, Branford S. Molecular monitoring of BCR-ABL as a guide to clinical management in chronic myeloid leukaemia. Blood Rev 2006; 20: 29–41

    Article  PubMed  CAS  Google Scholar 

  9. Landstrom AP, Tefferi A. Fluorescent in situ hybridization in the diagnosis, prognosis, and treatment monitoring of chronic myeloid leukemia. Leuk Lymph 2006; 47: 397–402

    Article  CAS  Google Scholar 

  10. Max N, Wolf K, Wittor B, et al. Rapid real-time quantification of BCR-ABL transcripts in research samples: intra- and inter-assay variation of a novel RT-PCR assay [abstract H13]. J Mol Diag 1999; 1: 48

    Google Scholar 

  11. SAS Institute Inc. SAS OnlineDoc® 9.1.3. Cary (NC): SAS Institute Inc., 2004

  12. Cortes J, Talpaz M, O’Brien S, et al. Molecular responses in patients with chronic myelogenous leukemia in chronic phase treated with imatinib mesylate. Clin Cancer Res 2005; 11: 3425–32

    Article  PubMed  CAS  Google Scholar 

  13. Tchirkov A, Giollant M, Tavernier F, et al. Interphase cytogenetics and competitive RT-PCR for residual disease monitoring with chronic myeloid leukaemia during interferon-α therapy. Brit J Haematol 1998; 101: 552–7

    Article  CAS  Google Scholar 

  14. Kim YJ, Kim DW, Lee S, et al. Comprehensive comparison of FISH, RT-PCR, and RQ-PCR for monitoring the BCR-ABL gene after hematopoietic stem cell transplantation in CML. Eur J Haematol 2002; 68: 272–80

    Article  PubMed  CAS  Google Scholar 

  15. Raanani P, Ben-Bassat I, Gan S, et al. Assessment of the response to imatinib in chronic myeloid leukemia patients: comparison between the FISH, multiplex and RT-PCR methods. Eur J Haematol 2002; 73: 243–50

    Article  Google Scholar 

  16. 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 2002; 16: 53–9

    Article  PubMed  CAS  Google Scholar 

  17. Bhatia R, Holtz M, Niu N, et al. Persistence of malignant hematopoietic progenitors in chronic myelogenous leukemia patients in complete cytogenetic remission following imatinib mesylate treatment. Blood 2003; 101: 4701–7

    Article  PubMed  CAS  Google Scholar 

  18. 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 interferon-α therapy or allogeneic bone marrow transplantation. Blood 2000; 95: 404–9

    PubMed  CAS  Google Scholar 

  19. Fuggazza G, Miglino M, Bruzzone R, et al. Cytogenetic and fluorescence in situ monitoring in Ph+ chronic myeloid leukemia patients treated with imatinib mesylate. J Exp Clin Cancer Res 2004; 23: 295–9

    Google Scholar 

  20. Hughes T. ABL kinase inhibitor therapy for CML: baseline assessments and response monitoring. Hematology Am Soc Hematol Educ Program 2006, 211-8

  21. Rulcová J, Zmeková V, Zemanová Z, et al. The effect of total-ABL, GUS and B2M control genes on BCR-ABL monitoring by real-time RT-PCR. Leuk Res 2007; 31: 483–91

    Article  PubMed  Google Scholar 

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Acknowledgments

The authors thank all colleagues and referring physicians for clinical information. The authors have no conflict of interest directly relevant to the content of this article.

This study was supported by research funds of the Feist Weiller Cancer Center.

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Authors and Affiliations

  1. Department of Pathology, Louisiana State University, Shreveport, Louisiana, USA

    Fei Bao, Clarissa Lowery, Sherry Martin, Diana M. Veillon, James D. Cotelingam & Mary Lowery Nordberg

  2. Division of Hematology/Oncology, (Feist Weiller Cancer Center), Department of Medicine, Louisiana State University Health Sciences Center, 1501 Kings Highway, Shreveport, LA, 71130, USA

    Reinhold Munker, Runhua Shi & Mary Lowery Nordberg

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  1. Fei Bao
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  2. Reinhold Munker
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  3. Clarissa Lowery
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  4. Sherry Martin
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  5. Runhua Shi
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  6. Diana M. Veillon
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  7. James D. Cotelingam
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Correspondence to Reinhold Munker.

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Bao, F., Munker, R., Lowery, C. et al. Comparison of FISH and Quantitative RT-PCR for the Diagnosis and Follow-Up of BCR-ABL-Positive Leukemias. Mol Diag Ther 11, 239–245 (2007). https://doi.org/10.1007/BF03256245

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