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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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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DOI: https://doi.org/10.1038/sj.leu.2404983
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