Abstract
In this chapter, selected molecular methods currently widely applied for the diagnostics and assessment of minimal residual disease (MRD) in patients with chronic myelogenous leukemia (CML) are presented. At the time of diagnosis, the qualitative RT-PCR method is employed. It allows for the identification of the type of BCR-ABL fusion gene. During follow-up, the quantitative RQ-PCR analysis is used for monitoring the kinetics of changes in the BCR-ABL fusion gene expression level. The results of these analyses might indicate the need for therapy modification. In case of undetectable BCR-ABL fusion gene levels in RQ-PCR, a more sensitive approach, nested RT-PCR, might be performed. In case of lack or loss of molecular response, the assessment of mutational status of ABL kinase domain is required.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Borrow J (2007) Guidelines for mutation analysis of BCR/ABL kinase domain: interpreting TKI-resistance mutations in CML patients. West Midlands Regional Genetics Laboratory. Available via http://www.bwhct.nhs.uk/tki_sensitivities_final.pdf. Cited 9 Dec 2011
Branford S, Hughes T (2006) Detection of BCR-ABL mutations and resistance to imatinib mesylate. Methods Mol Med 125:93–106
Branford S, Cross NC, Hochhaus A, Radich J, Saglio G, Kaeda J, Goldman J, Hughes T (2006) Rationale for the recommendations for harmonizing current methodology for detecting BCR-ABL transcripts in patients with chronic myeloid leukaemia. Leukemia 20:1925–1930
Ernst T, Hoffmann J, Erben P, Hanfstein B, Leitner A, Hehlmann R, Hochhaus A, Müller MC (2008) ABL single nucleotide polymorphisms may masquerade as BCR-ABL mutations associated with resistance to tyrosine kinase inhibitors in patients with chronic myeloid leukemia. Haematologica 93:1389–1393
Gabert J, Beillard E, van der Velden VH, Bi W, Grimwade D, Pallisgaard N, Barbany G, Cazzaniga G, Cayuela JM, Cavé H, Pane F, Aerts JL, De Micheli D, Thirion X, Pradel V, González M, Viehmann S, Malec M, Saglio G, van Dongen JJ (2003) Standardization and quality control studies of ‘real-time’ quantitative reverse transcriptase polymerase chain reaction of fusion gene transcripts for residual disease detection in leukemia—a Europe Against Cancer program. Leukemia 17:2318–2357
Hughes T, Branford S (2006) Molecular monitoring of BCR-ABL as a guide to clinical management in chronic myeloid leukaemia. Blood Rev 20:29–41
Hughes T, Branford S (2009) Monitoring disease response to tyrosine kinase inhibitor therapy in CML. Hematol Am Soc Hematol Educ Program 477-87
Hughes T, Deininger M, Hochhaus A, Branford S, Radich J, Kaeda J, Baccarani M, Cortes J, Cross NC, Druker BJ, Gabert J, Grimwade D, Hehlmann R, Kamel-Reid S, Lipton JH, Longtine J, Martinelli G, Saglio G, Soverini S, Stock W, Goldman JM (2006) Monitoring CML patients responding to treatment with tyrosine kinase inhibitors: review and recommendations for harmonizing current methodology for detecting BCR-ABL transcripts and kinase domain mutations and for expressing results. Blood 108:28–37
Jones D, Kamel-Reid S, Bahler D, Dong H, Elenitoba-Johnson K, Press R, Quigley N, Rothberg P, Sabath D, Viswanatha D, Weck K, Zehnder J (2009) Laboratory practice guidelines for detecting and reporting BCR-ABL drug resistance mutations in chronic myelogenous leukemia and acute lymphoblastic leukemia: a report of the Association for Molecular Pathology. J Mol Diagn 11:4–11
Martinelli G, Soverini S, Rosti G, Cilloni D, Baccarani M (2005) New tyrosine kinase inhibitors in chronic myeloid leukemia. Haematologica 90:534–541
Müller MC, Cross NC, Erben P, Schenk T, Hanfstein B, Ernst T, Hehlmann R, Branford S, Saglio G, Hochhaus A (2009) Harmonization of molecular monitoring of CML therapy in Europe. Leukemia 23:1957–1963
Shah NP, Sawyers CL (2003) Mechanisms of resistance to STI571 in Philadelphia chromosome-associated leukemias. Oncogene 20(22):7389–7395
Stentoft J, Pallisgaard N, Kjeldsen E, Holm MS, Nielsen JL, Hokland P (2001) Kinetics of BCR-ABL fusion transcript levels in chronic myeloid leukemia patients treated with STI571 measured by quantitative real- time polymerase chain reaction. Eur J Haematol 67:302–308
van Dongen JJ, Macintyre EA, Gabert JA, Delabesse E, Rossi V, Saglio G, Gottardi E, Rambaldi A, Dotti G, Griesinger F, Parreira A, Gameiro P, Diáz MG, Malec M, Langerak AW, San Miguel JF, Biondi A (1999) Standardized RT-PCR analysis of fusion gene transcripts from chromosome aberrations in acute leukaemia for detection of minimal residual disease. Report of the BIOMED-1 Concerted Action: investigation of minimal residual disease in acute leukaemia. Leukemia 13:1901–1928
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2012 Springer-Verlag Berlin Heidelberg
About this chapter
Cite this chapter
Czekalska, S., Zawada, M., Florek, I. (2012). Assessment of the Presence and the Level of BCR-ABL Fusion Gene Expression and Mutational Status in ABL Kinase Domain. In: Witt, M., Dawidowska, M., Szczepanski, T. (eds) Molecular Aspects of Hematologic Malignancies. Principles and Practice. Springer, Berlin, Heidelberg. https://doi.org/10.1007/978-3-642-29467-9_25
Download citation
DOI: https://doi.org/10.1007/978-3-642-29467-9_25
Published:
Publisher Name: Springer, Berlin, Heidelberg
Print ISBN: 978-3-642-29466-2
Online ISBN: 978-3-642-29467-9
eBook Packages: Biomedical and Life SciencesBiomedical and Life Sciences (R0)