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  • Biotechnical Methods Section BTS
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Biotechnical Methods Section (BTS)

Quality assurance in RT-PCR-based BCR/ABL diagnostics – results of an interlaboratory test and a standardization approach

Leukemia volume 14, pages 1850–1856 (2000)Cite this article

Abstract

Here we describe the results of an interlaboratory test for RT-PCR-based BCR/ABL analysis. The test was organized in two parts. The number of participating laboratories in the first and second part was 27 and 20, respectively. In the first part samples containing various concentrations of plasmids with the ela2, b2a2 or b3a2 BCR/ABL transcripts were analyzed by PCR. In the second part of the test, cell samples containing various concentrations of BCR/ABL-positive cells were analyzed by RT-PCR. Overall PCR sensitivity was sufficient in approximately 90% of the tests, but a significant number of false positive results were obtained. There were significant differences in sensitivity in the cell-based analysis between the various participants. The results are discussed, and proposals are made regarding the choice of primers, controls, conditions for RNA extraction and reverse transcription.

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References

  1. Maurer J, Janssen JWG, Thiel E, Van Denderen J, Ludwig W-D, Aydemir Ü, Heinze B, Fonatsch C, Harbott J, Reiter A, Riehm H, Hoelzer D, Bartram CR . Detection of chimeric BCR/ABL genes in acute lymphoblastic leukemia by the polymerase chain reaction Lancet 1991 337: 1055–1058

    Article  CAS  Google Scholar 

  2. Schlieben S, Borkhardt A, Reinisch I, Ritterbach J, Janssen JWG, Ratei R, Schrappe M, Repp R, Zimmermann M, Kabisch H, Janka-Schaub G, Bartram CR, Ludwig W-D, Lampert F, Harbott J . Incidence and clinical outcome of children with BCR/ABL-positive acute lymphoblastic leukemia (ALL). A prospective RT-PCR study based on 673 patients enrolled in the German pediatric multicenter therapy trials ALL-BFM-90 and CoALL-05-92 Leukemia 1996 10: 957–963

    CAS  PubMed  Google Scholar 

  3. Hoelzer D, Gökbuget N, Arnold R, Büchner T, Freund M, Gassmann W, Heil G, Hiddemann W, Löffler H, Lipp T, Ludwig WD, Maschmeyer G, Thiel E, Messerer D . Acute lymphatic leukemia of the adult. Diagnosis, risk groups and therapy Internist (Berl) 1997 37: 994–1007

    Google Scholar 

  4. Heisterkamp N, Stam K, Groffen J, de Klein A, Grosveld G . Structural organization of the bcr gene and its role in the Ph′ translocation Nature 1985 315: 758–761

    Article  CAS  Google Scholar 

  5. Pane F, Frigeri F, Sindona M, Luciano L, Ferrara F, Cimino R, Meloni G, Saglio G, Salvatore F, Rotoli B . Neutrophilic-chronic myeloid leukemia: a distinct disease with a specific molecular marker (BCR/ABL with C3/A2 junction) Blood 1996 88: 2410–2414

    CAS  PubMed  Google Scholar 

  6. How GF, Tan LT, Lim LC . Chronic myeloid leukemia with e19a2 (c3a2) BCR/ABL fusion junction – is it truly a benign disease? Leukemia 1998 12: 1166–1167

    Article  CAS  Google Scholar 

  7. Melo JV . BCR-ABL gene variants Baillières Clin Haematol 1997 10: 203–222

    Article  CAS  Google Scholar 

  8. Iwata S, Mizutani S, Nakazawa S, Yata J . Heterogeneity of the breakpoint in the ABL gene in cases with BCR/ABL transcript lacking ABL exon a2 Leukemia 1994 8: 1696–1702

    CAS  PubMed  Google Scholar 

  9. Lozzio CB, Lozzio BB . Human chronic myelogenous leukemia cell line with positive Philadelphia chromosome Blood 1975 45: 321–334

    CAS  PubMed  Google Scholar 

  10. Seelig R, Schranz P, Bottner C, Neubauer A, Seelig HP, Renz M . The chronic myelocytic cell line K 562 contains minor (m) as well as major (M) bcr/abl fusion mRNAs Leukemia 1993 7: 1886–1887

    CAS  PubMed  Google Scholar 

  11. Pegoraro L, Matera L, Ritz J, Levis A, Palumbo A, Biagini G . Establishment of a Ph1-positive human cell line (BV173) J Natl Cancer Inst 1983 70: 447–453

    CAS  PubMed  Google Scholar 

  12. van Rhee F, Hochhaus A, Lin F, Melo JV, Goldman JM, Cross NCP . p190 BCR-ABL mRNA is expressed at low levels in p210-positive chronic myeloid and acute lymphoblastic leukemia Blood 1996 87: 5213–5217

    CAS  PubMed  Google Scholar 

  13. Dhut S, Gibbons B, Chaplin T, Young BD . Establishment of a lymphoblastoid cell line, SD-1, expressing the p190 bcr-abl chimaeric protein Leukemia 1991 5: 49–55

    CAS  PubMed  Google Scholar 

  14. Cross NC, Melo JV, Feng L, Goldman JM . An optimized multiplex polymerase chain reaction (PCR) for detection of BCR-ABL fusion mRNAs in haematological disorders Leukemia 1994 8: 186–189

    CAS  PubMed  Google Scholar 

  15. Lee A, Kirk J, Edmands S, Radich J . Multiplex PCR of bcr-abl fusion transcripts in Philadelphia positive acute lymphoblastic leukemia PCR Methods Appl 1995 4: 283–287

    Article  CAS  Google Scholar 

  16. Hochhaus A, Reiter A, Skladny H, Melo JV, Sick C, Berger U, Guo JQ, Arlinghaus RB, Hehlmann R, Goldman JM, Cross NC . A novel BCR-ABL fusion gene (e6a2) in a patient with Philadelphia chromosome-negative chronic myelogenous leukemia Blood 1996 88: 2236–2240

    CAS  PubMed  Google Scholar 

  17. Kidd V, Lion T . Debate round table: appropriate controls for RT-PCR Leukemia 1997 11: 871–881

    Article  CAS  Google Scholar 

  18. van Dongen JJ, Seriu T, Panzer-Grumayer ER, Biondi A, Pongers-Willemse MJ, Corral L, Stolz F, Schrappe M, Masera G, Kamps WA, Gadner H, van Wering ER, Ludwig WD, Basso G, de Bruijn MA, Cazzaniga G, Hettinger K, van der Does-van den Berg A, Hop WC, Riehm H, Bartram CR . Prognostic value of minimal residual disease in acute lymphoblastic leukaemia in childhood Lancet 1998 352: 1731–1738

    Article  CAS  Google Scholar 

  19. 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, Diaz MG, Malec M, Langerak AW, San Miguel JF, Biondi A . Standardized RT-PCR analysis of fusion gene transcripts from chromosome aberrations in acute leukemia for detection of minimal residual disease. Report of the BIOMED-1 Concerted Action: investigation of minimal residual disease in acute leukemia Leukemia 1999 13: 1901–1928

    Article  CAS  Google Scholar 

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Acknowledgements

This work was supported by a grant from the German Federal Ministry of Health (Bundesministerium für Gesundheit), project ‘quality assurance in molecular medicine’. The authors (TB and ET) wish to thank all participants for their cooperation.

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Authors and Affiliations

  1. Freie Universität Berlin, Med Klinik III, Germany

    T Burmeister, J Maurer & E Thiel

  2. Universität Düsseldorf, Klinik für Hämatologie und Onkologie, Germany

    M Aivado

  3. Universität-Gesamthochschule Essen, Klinik für Knochenmarktransplantation, Germany

    AH Elmaagacli

  4. Universität Tübingen, Med, Klinik, Germany

    F Grünebach

  5. Praxis Held/Kesser/Arndt Hamburg, Germany

    KR Held

  6. Universität Mainz, III, Med, Klinik, Germany

    G Heß

  7. Universität Heidelberg, III, Med, Klinik, Klinikum Mannheim, Germany

    A Hochhaus

  8. GLE medicon GmbH, Hamburg, Germany

    W Höppner

  9. Universität Trier, Abt, Molekulare Neurogenetik, Germany

    KU Lentes

  10. Universität Freiburg, Med. Klinik I, Germany

    M Lübbert

  11. Universität Münster, Institut für Pathologie, Germany

    KL Schäfer

  12. Universität Hamburg, Med. Klinik III, Germany

    P Schafhausen

  13. Humboldt-Universität Berlin, Med. Klinik, Hämatologie/Onkologie, Germany

    CA Schmidt

  14. Universität Greifswald, Klinik für Innere Medizin C, Germany

    F Schüler

  15. Humboldt-Universität Berlin, Zentrum für Kinderheilkunde, Germany

    K Seeger

  16. Labor Prof Seelig und Partner, Karlsruhe, Germany

    R Seelig

  17. Technische Universität Dresden, Med, Klinik I, Germany

    C Thiede

  18. Universität Gießen, Kinderklinik, Onkogenetisches Labor, Germany

    S Viehmann

  19. Universität Gießen, Abt, für Hämatologie, Germany

    C Weber

  20. Universität Rostock, Abt für Hämatologie/Onkologie, Germany

    S Wilhelm

  21. Universität Homburg, Institut für Humangenetik, Germany

    A Christmann

  22. Universität Jena, Innere Medizin II, Germany

    JH Clement

  23. Universität Frankfurt, Zentrum für Kinderheilkunde, Germany

    U Ebener

  24. Universität Düsseldorf, Knochenmarkspenderzentrale, Germany

    J Enczmann

  25. Universität Münster, Med, Klinik A, Germany

    R Leo

  26. LMU München, Klinikum Groβhadern, Med, Klinik III, Germany

    M Schleuning

  27. Universität Kiel, II, Med, Klinik, Germany

    R Schoch

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  1. T Burmeister
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  2. J Maurer
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  14. CA Schmidt
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  15. F Schüler
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  17. R Seelig
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  18. C Thiede
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  19. S Viehmann
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  20. C Weber
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  21. S Wilhelm
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  29. E Thiel
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Burmeister, T., Maurer, J., Aivado, M. et al. Quality assurance in RT-PCR-based BCR/ABL diagnostics – results of an interlaboratory test and a standardization approach. Leukemia 14, 1850–1856 (2000). https://doi.org/10.1038/sj.leu.2401899

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  • DOI: https://doi.org/10.1038/sj.leu.2401899

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