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Clinical Chemistry and Laboratory Medicine (CCLM)

Published in Association with the European Federation of Clinical Chemistry and Laboratory Medicine (EFLM)

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Ed. by Gillery, Philippe / Lackner, Karl J. / Lippi, Giuseppe / Melichar, Bohuslav / Payne, Deborah A. / Schlattmann, Peter / Tate, Jillian R.

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Volume 44, Issue 5 (May 2006)


Real-time RT-PCR quantification of PRAME gene expression for monitoring minimal residual disease in acute myeloblastic leukaemia

Nicolas Tajeddine / Isabelle Millard
  • Laboratory of Applied Molecular Technology, Center for Human Genetics, Université catholique de Louvain, Brussels, Belgium
  • Other articles by this author:
  • De Gruyter OnlineGoogle Scholar
/ Philippe Gailly / Jean-Luc Gala
  • Laboratory of Applied Molecular Technology, Center for Human Genetics, Université catholique de Louvain, Brussels, Belgium and Defence Laboratories Department (DG MR), Belgian Armed Forces, Brussels, Belgium
  • Other articles by this author:
  • De Gruyter OnlineGoogle Scholar
Published Online: 2011-09-21 | DOI: https://doi.org/10.1515/CCLM.2006.106


Background: Specific gene rearrangements are used for minimal residual disease (MRD) assessment, but are frequently lacking in leukaemias. In these cases, the quantification of PRAME (preferentially expressed antigen of melanoma) transcripts could be useful.

Methods: PRAME transcripts were quantified by real-time RT-PCR in normal and leukaemic samples, and the results were compared with those of conventional RT-PCR. Basal expression of PRAME was determined in 25 blood samples and 25 bone marrow samples from healthy donors, as well as in 12 haematological cell lines (Jurkat, K562, HL60, DOHH2, IM9, Daudi, CEM, KG1, DG75, 8226, U937, Raji).

Results: In paediatric acute myeloid leukaemia (AML) (n=22) and acute lymphoblastic leukaemia (ALL) (n=17), and in adult AML (n=20), abnormal PRAME expression was found in 41%, 35% and 40% of cases, respectively. To assess the sensitivity of PRAME for monitoring MRD, PRAME-positive t(8;21) AML samples with detectable AML1/ETO expression by conventional RT-PCR (n=17) were assessed for quantitative expression of AML1/ETO and PRAME. The expression of these genes was closely correlated. To confirm that PRAME expression was correlated with clinical data, the expression of PRAME was also sequentially followed in patients (n=13) from onset to cytological remission or relapse. The cytological and molecular data were highly correlated in all patients.

Conclusions: Our data confirm that PRAME quantification by real-time RT-PCR appears suitable for monitoring MRD in PRAME-positive leukaemia.

Keywords: leukaemia; minimal residual disease; PRAME (preferentially expressed antigen of melanoma); real-time RT-PCR


  • 1.

    Campana D. Determination of minimal residual disease in leukaemia patients. Br J Haematol 2003; 121:823–38.Google Scholar

  • 2.

    Cave H, van der Werff ten Bosch J, Suciu S, Guidal C, Waterkeyn C, Otten J, et al. Clinical significance of minimal residual disease in childhood acute lymphoblastic leukemia. European Organization for Research and Treatment of Cancer – Childhood Leukemia Cooperative Group. N Engl J Med 1998; 339:591–8.Google Scholar

  • 3.

    Pongers-Willemse MJ, Verhagen OJ, Tibbe GJ, Wijkhuijs AJ, de Haas V, Roovers E, et al. Real-time quantitative PCR for the detection of minimal residual disease in acute lymphoblastic leukemia using junctional region specific TaqMan probes. Leukemia 1998; 12:2006–14.CrossrefGoogle Scholar

  • 4.

    van Dongen JJ, Seriu T, Panzer-Grumayer ER, Biondi A, Pongers-Willemse MJ, Corral L, et al. Prognostic value of minimal residual disease in acute lymphoblastic leukaemia in childhood. Lancet 1998; 352:1731–8.Google Scholar

  • 5.

    Ikeda H, Lethe B, Lehmann F, van Baren N, Baurain JF, de Smet C, et al. Characterization of an antigen that is recognized on a melanoma showing partial HLA loss by CTL expressing an NK inhibitory receptor. Immunity 1997; 6:199–208.CrossrefGoogle Scholar

  • 6.

    van Baren N, Chambost H, Ferrant A, Michaux L, Ikeda H, Millard I, et al. PRAME, a gene encoding an antigen recognized on a human melanoma by cytolytic T cells, is expressed in acute leukaemia cells. Br J Haematol 1998; 102:1376–9.Google Scholar

  • 7.

    Tajeddine N, Gala JL, Louis M, Van Schoor M, Tombal B, Gailly P. Tumor-associated antigen preferentially expressed antigen of melanoma (PRAME) induces caspase-independent cell death in vitro and reduces tumorigenicity in vivo. Cancer Res 2005; 65:7348–55.Google Scholar

  • 8.

    McElwaine S, Mulligan C, Groet J, Spinelli M, Rinaldi A, Denyer G, et al. Microarray transcript profiling distinguishes the transient from the acute type of megakaryoblastic leukaemia (M7) in Down's syndrome, revealing PRAME as a specific discriminating marker. Br J Haematol 2004; 125:729–42.Google Scholar

  • 9.

    Steinbach D, Hermann J, Viehmann S, Zintl F, Gruhn B. Clinical implications of PRAME gene expression in childhood acute myeloid leukemia. Cancer Genet Cytogenet 2002; 133:118–23.Google Scholar

  • 10.

    Matsushita M, Yamazaki R, Ikeda H, Kawakami Y. Preferentially expressed antigen of melanoma (PRAME) in the development of diagnostic and therapeutic methods for hematological malignancies. Leuk Lymphoma 2003; 44:439–44.CrossrefGoogle Scholar

  • 11.

    Matsushita M, Ikeda H, Kizaki M, Okamoto S, Ogasawara M, Ikeda Y, et al. Quantitative monitoring of the PRAME gene for the detection of minimal residual disease in leukaemia. Br J Haematol 2001; 112:916–26.Google Scholar

  • 12.

    Guerrasio A, Pilatrino C, De Micheli D, Cilloni D, Serra A, Gottardi E, et al. Assessment of minimal residual disease (MRD) in CBFbeta/MYH11-positive acute myeloid leukemias by qualitative and quantitative RT-PCR amplification of fusion transcripts. Leukemia 2002; 16:1176–81.CrossrefGoogle Scholar

  • 13.

    Wattjes MP, Krauter J, Nagel S, Heidenreich O, Ganser A, Heil G. Comparison of nested competitive RT-PCR and real-time RT-PCR for the detection and quantification of AML1/MTG8 fusion transcripts in t(8;21) positive acute myelogenous leukemia. Leukemia 2000; 14:329–35.Google Scholar

  • 14.

    Donovan JW, Ladetto M, Zou G, Neuberg D, Poor C, Bowers D, et al. Immunoglobulin heavy-chain consensus probes for real-time PCR quantification of residual disease in acute lymphoblastic leukemia. Blood 2000; 95:2651–8.Google Scholar

  • 15.

    Cilloni D, Gottardi E, De Micheli D, Serra A, Volpe G, Messa F, et al. Quantitative assessment of WT1 expression by real time quantitative PCR may be a useful tool for monitoring minimal residual disease in acute leukemia patients. Leukemia 2002; 16:2115–21.CrossrefGoogle Scholar

  • 16.

    Hosen N, Sonoda Y, Oji Y, Kimura T, Minamiguchi H, Tamaki H, et al. Very low frequencies of human normal CD34+ haematopoietic progenitor cells express the Wilms' tumour gene WT1 at levels similar to those in leukaemia cells. Br J Haematol 2002; 116:409–20.Google Scholar

  • 17.

    Elmaagacli AH, Beelen DW, Trenschel R, Schaefer UW. The detection of wt-1 transcripts is not associated with an increased leukemic relapse rate in patients with acute leukemia after allogeneic bone marrow or peripheral blood stem cell transplantation. Bone Marrow Transplant 2000; 25:91–6.CrossrefGoogle Scholar

  • 18.

    Steinbach D, Viehmann S, Zintl F, Gruhn B. PRAME gene expression in childhood acute lymphoblastic leukemia. Cancer Genet Cytogenet 2002; 138:89–91.Google Scholar

  • 19.

    Paydas S, Tanriverdi K, Yavuz S, Disel U, Baslamisli F, Burgut R. PRAME mRNA levels in cases with acute leukemia: clinical importance and future prospects. Am J Hematol 2005; 79:257–61.Google Scholar

  • 20.

    Deindl E, Boengler K, van Royen N, Schaper W. Differential expression of GAPDH and beta3-actin in growing collateral arteries. Mol Cell Biochem 2002; 236:139–46.Google Scholar

  • 21.

    Glare EM, Divjak M, Bailey MJ, Walters EH. β-Actin and GAPDH housekeeping gene expression in asthmatic airways is variable and not suitable for normalising mRNA levels. Thorax 2002; 57:765–70.Google Scholar

  • 22.

    Selvey S, Thompson EW, Matthaei K, Lea RA, Irving MG, Griffiths LR. β-Actin – an unsuitable internal control for RT-PCR. Mol Cell Probes 2001; 15:307–11.CrossrefGoogle Scholar

  • 23.

    Steele BK, Meyers C, Ozbun MA. Variable expression of some “housekeeping” genes during human keratinocyte differentiation. Anal Biochem 2002; 307:341–7.Google Scholar

  • 24.

    Gala JL, Heusterspreute M, Loric S, Hanon F, Tombal B, Van Cangh P, et al. Expression of prostate-specific antigen and prostate-specific membrane antigen transcripts in blood cells: implications for the detection of hematogenous prostate cells and standardization. Clin Chem 1998; 44:472–81.Google Scholar

  • 25.

    Dekairelle A, Tombal B, Cosyns J, Gala J. Assessment of the transcriptional activity of p53 improves the prediction of recurrence in superficial transitional cell carcinoma (TCC) of the bladder. Clin Cancer Res 2005; 11:4724–32.Google Scholar

  • 26.

    Greiner J, Ringhoffer M, Simikopinko O, Szmaragowska A, Huebsch S, Maurer U, et al. Simultaneous expression of different immunogenic antigens in acute myeloid leukemia. Exp Hematol 2000; 28:1413–22.Google Scholar

  • 27.

    Greiner J, Ringhoffer M, Taniguchi M, Li L, Schmitt A, Shiku H, et al. mRNA expression of leukemia-associated antigens in patients with acute myeloid leukemia for the development of specific immunotherapies. Int J Cancer 2004; 108:704–11.Google Scholar

About the article

Corresponding author: Prof. Jean-Luc Gala, MD, PhD, Laboratory of Applied Molecular Technology, Center for Human Genetics (UCL), Clos Chappelle-aux-Champs, 30-UCL30.46, 1200 Brussels, Belgium Phone: +32-2-7643165, Fax: +32-2-7643166,

Received: 2005-12-14

Accepted: 2006-02-12

Published Online: 2011-09-21

Published in Print: 2006-05-01

Citation Information: Clinical Chemistry and Laboratory Medicine (CCLM), ISSN (Online) 1437-4331, ISSN (Print) 1434-6621, DOI: https://doi.org/10.1515/CCLM.2006.106.

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©2006 by Walter de Gruyter Berlin New York. Copyright Clearance Center

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