Primers and protocols for standardized detection of minimal residual disease in acute lymphoblastic leukemia using immunoglobulin and T cell receptor gene rearrangements and TAL1 deletions as PCR targets Report of the BIOMED-1 CONCERTED ACTION: Investigation of minimal residual disease in acute leukemia (original) (raw)

Biotechnical Methods Section (BTS)

Leukemia volume 13, pages 110–118 (1999)Cite this article

Abstract

It is now widely accepted that the detection of minimal residual disease (MRD) has prognostic value in acute leukemia. However clinical MRD studies need standardized techniques. Therefore, several European laboratories have aligned their goals and performed comparative studies to achieve optimization and standardization of MRD techniques. This was achieved via the BIOMED-1 Concerted Action “Investigation of minimal residual disease in acute leukemia: International standardization and clinical evaluation.” This report describes the development of PCR primers and protocols for the detection of MRD in acute lymphoblastic leukemia (ALL) using clone-specific junctional regions of immunoglobulin and T cell receptor gene rearrangements and TAL1 deletions as PCR targets. A total of 54 primers was developed (1) to amplify rearrangements of the TCRD, TCRG, and IGK (Kde) genes as well as TAL1 deletions; (2) to sequence the junctional regions and breakpoint fusion regions; and (3) to perform MRD detection in bone marrow or peripheral blood samples during follow-up of ALL patients. Protocols were established to identify PCR targets at diagnosis by performing 25 PCR reactions per patient using appropriate positive and negative controls. Standardized protocols were developed for MRD monitoring via single amplification of the PCR target followed by dot blot hybridization with the corresponding patient-specific junctional region probe. In addition, alternative approaches were designed for cases where the target sensitivity of at least 10−4 was not obtained. The standardization described here of MRD-PCR techniques is essential for the process of translating MRD research into clinical practice.

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Authors and Affiliations

  1. Department of Immunology, University Hospital Rotterdam/Erasmus University Rotterdam, Rotterdam, The Netherlands
    MJ Pongers-Willemse & JJM van Dongen
  2. Institute of Human Genetics, University of Heidelberg, Heidelberg, Germany
    T Seriu & CR Bartram
  3. Children’s Cancer Research Institute, St Anna Kinderspital, Vienna, Austria
    F Stolz & ER Panzer-Grümayer
  4. Department of Pediatrics, Universitá Milano, Ospedale San Gerardo, Monza, Italy
    E d’Aniello & A Biondi
  5. Departamento de Hematologia, Instituto Português de Oncologia, Lisboa, Portugal
    P Gameiro
  6. Department of Hematology, Karolinska Hospital, Stockholm, Sweden
    P Pisa
  7. Department of Hematology, Hospital Clinico Universitario, Salamanca, Spain
    M Gonzalez & JF San Miguel

Authors

  1. MJ Pongers-Willemse
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  2. T Seriu
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  3. F Stolz
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  4. E d’Aniello
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  5. P Gameiro
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  6. P Pisa
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  7. M Gonzalez
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  8. CR Bartram
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  9. ER Panzer-Grümayer
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  10. A Biondi
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  11. JF San Miguel
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  12. JJM van Dongen
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Pongers-Willemse, M., Seriu, T., Stolz, F. et al. Primers and protocols for standardized detection of minimal residual disease in acute lymphoblastic leukemia using immunoglobulin and T cell receptor gene rearrangements and TAL1 deletions as PCR targets Report of the BIOMED-1 CONCERTED ACTION: Investigation of minimal residual disease in acute leukemia.Leukemia 13, 110–118 (1999). https://doi.org/10.1038/sj.leu.2401245

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