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Chronic Myeloproliferative Disorders

Harmonization of BCR-ABL mRNA quantification using a uniform multifunctional control plasmid in 37 international laboratories

Leukemia volume 22, pages 96–102 (2008)Cite this article

Abstract

Individualized PCR strategies hamper comparability of molecular results between different laboratories in several fields of medicine. To harmonize BCR-ABL mRNA quantification an international multicenter trial involving 37 laboratories in 14 countries was initiated using 10 samples, each containing various dilutions (10, 2, 1 and 0.1%) of b3a2 or b2a2 BCR-ABL positive in normal leukocytes and negative controls. A novel control plasmid (pME-2) was designed for external calibration containing BCR-ABL and glucuronidase-β (GUS) sequences. Median BCR-ABL/ABL ratios were 9.1, 1.8, 0.85 and 0.11% in b3a2 samples and 9.5, 1.6, 0.84 and 0.11% in b2a2 samples. Median BCR-ABL/GUS ratios were 3.4, 0.77, 0.37 and 0.042% in b3a2 samples and 2.8, 0.48, 0.29 and 0.031% in b2a2 samples. The coefficients of variation were 0.62 for ratios BCR-ABL/ABL and 1.03 for ratios BCR-ABL/GUS. Five of 37 evaluable participating laboratories (13%) detected low BCR-ABL copy numbers in negative control samples; one laboratory failed to detect BCR-ABL in a low-level sample. We conclude that the use of a common control plasmid does indeed improve comparability of BCR-ABL mRNA quantification results. However, further standardizing efforts like introducing a calibrator and regular control rounds are needed.

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Acknowledgements

The study was supported by the Competence Network ‘Acute and chronic leukemias,’ sponsored by the German Bundesministerium für Bildung und Forschung (Projektträger Gesundheitsforschung; DLR e.V.- 01 GI9980/6), the European LeukemiaNet (WP 4+12) within the 6th European Framework Programme for Research and Technological Development, the German José Carreras-Foundation, Qiagen, Hilden, Germany, and Novartis Pharma, Nürnberg, Germany.

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Author notes

  1. M C Müller and P Erben: These authors contributed equally to this work.

Authors and Affiliations

  1. III. Medizinische Universitätsklinik, Medizinische Fakultät Mannheim der Universität Heidelberg, Mannheim, Germany

    M C Müller, P Erben, T Schenk, T Ernst, S Lauber, J Kruth, R Hehlmann & A Hochhaus

  2. Division of Hematology and Internal Medicine, Department of Clinical and Biological Sciences, University of Turin, Turin, Italy

    G Saglio & E Gottardi

  3. Department of Hematology, Aarhus University Hospital, Aarhus, Denmark

    C G Nyvold

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  1. M C Müller
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on behalf of the European LeukemiaNet

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Correspondence to M C Müller.

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Participating laboratories

Division of Hematology and Internal Medicine, Department of Clinical and Biological Sciences, University of Turin, Turin, Italy (Giuseppe Saglio, Enrico Gottardi); Klinisches Institut für Medizinische und Chemische Labordiagnostik, Ehemalige I. Medizinische Universitätsklinik, Vienna, Austria (Harald Esterbauer); Molecular Biology, Children's Cancer Research Institute, Vienna, Austria (Thomas Lion); Department of Molecular Genetics, Institute of Hematology and Blood Transfusion, Prague, Czech Republic (Jana Rulcova, Cedric Haskovec); Department of Hematology, Aarhus University Hospital, Aarhus, Denmark (Charlotte Guldborg Nyvold); Department of Pathology, Odense University Hospital, Odense, Denmark (Niels Pallisgaard); Laboratoire Central d'Hématologie, Hôpital Saint-Louis, Paris, France (Jean-Michel Cayuela); Luminy Biotech Entreprises, Marseille, France (Fabienne Hermitte); INSERM U 817, Institut de Recherche contre le Cancer, Lille, France (Claude Preudhomme); INSERM E 217, Université Victor Segalen, Bordeaux, France (Francois-Xavier Mahon); Laboratoire de cytogénétique et biologie moléculaire, Centre hospitalier Lyon sud, Pierre Bénite, France (Sandrine Hayette); Klinik für Hämatologie, Onkologie und klinische Immunologie, Universität Düsseldorf, Düsseldorf, Germany (Ralf Kronenwett); Zell- und Molekularpathologie, Medizinische Hochschule, Hannover, Germany (Nils von Neuhoff); MLL-Munich Leukemia Lab, München, Germany (Susanne Schnittger); II. Medizinische Klinik, Universität Leipzig, Leipzig, Germany (Thoralf Lange); III. Medizinische Klinik, Universität Ulm, Ulm, Germany (Konstanze Döhner); Medizinische Klinik und Poliklinik I, Universität Dresden, Dresden, Germany (Christian Thiede); III. Medizinische Klinik, Universität Mainz, Mainz, Germany (Georg Heß); Klinik für Innere Medizin C, Universtität Greifswald, Greifswald, Germany (Frank Schüler); Institute of Hematology and Medical Oncology, University of Bologna, Bologna, Italy (Angela Poerio, Giovanni Martinelli); Department of Hematology, University of Naples, Naples, Italy (Fabrizio Pane); Department of Hematology, Jagiellonian University, Cracow, Poland (Tomasz Sacha); Department of Hematology at Hospital de la Santa Creu i Sant Pau, Universitat Autonoma de Barcelona, Barcelona, Spain (Josep Nomdedeu); Department of Hematopathology, University of Barcelona, Hospital Clinic, Barcelona, Spain (Dolors Colomer); Hematology Department, Hospital Universitario de Salamanca, Salamanca, Spain (Marcos Gonzalez Diaz); Laboratorio de Biologia Molecular, Hospital Universitario La Fe, Valencia, Spain (Pascual Bolufer); Molecular Medicine and Surgery, Karolinska Hospital, Stockholm, Sweden (Gisela Barbany); Molekulare Diagnostik, Inselspital, Bern, Switzerland (Elisabeth Oppliger-Leibundgut); Department of Hematology, VU University Medical Center, Amsterdam, The Netherlands (Quinten Waisfisz); Department of Genetics, Istanbul University , Istanbul, Turkey (Ugur Ozbek); Department of Clinical Haematology, Manchester Royal Infirmary, Manchester, United Kingdom (Abida Awan); Molecular Haematology, Glasgow Royal Infirmary, Glasgow, United Kingdom (Fiona Reid, Jan Baird); Department of Haematology, University of Liverpool, Liverpool, United Kingdom (Richard E Clark); II. Medizinische Klinik, Universität Frankfurt, Germany (Heike Pfeifer); Department of Pathology, Oregon Health and Science University, Portland OR, USA (Richard D Press); Department of Clinical Pathology, Cleveland Clinic Lerner College of Medicine, Cleveland, OH, USA (Raymond R Tubbs).

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Müller, M., Erben, P., Saglio, G. et al. Harmonization of BCR-ABL mRNA quantification using a uniform multifunctional control plasmid in 37 international laboratories. Leukemia 22, 96–102 (2008). https://doi.org/10.1038/sj.leu.2404983

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