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Digoxigenin-end labeled oligonucleotides facilitate the detection of minimal residual disease in acute lymphoblastic leukemia

Leukemia volume 14, pages 767–768 (2000)Cite this article

Recently, three large prospective trials unequivocally demonstrated that the evaluation of minimal residual disease (MRD) plays a pivotal role as a prognostic parameter in acute lymphoblastic leukemia (ALL).123 Individualized therapeutic adaptations based on the MRD status might be of considerable benefit for ALL patients. Several methodologies are available for MRD analyses in ALL patients.1234 Immunoglobulin (Ig) and T cell receptor (TCR) gene rearrangements have been utilized as PCR targets. An important PCR strategy for MRD analysis is the amplification of the rearranged Ig or TCR genes and hybridization with 32P-labeled clone-specific probes in dot blot hybridization. Now that the relevance of MRD monitoring in ALL has been recognized, modification of current techniques or development of new techniques enabling rapid and reliable MRD analysis is relevant. We report on a non-radioactive dot blot analysis taking advantage of digoxigenin (DIG)-labeled oligonucleotides for the rapid detection of MRD. DIG is a steroid hapten which is linked via a spacer arm to a nucleotide. DIG-ddUTP is incorporated at the 3′-end of an oligonucleotide by terminal transferase. After hybridization, the target DNA is detectable by an enzyme-linked immunoassay through an anti-DIG-AP Fab fragment.

We applied this technique on 10 adult ALL patients with IGK and/or TCRD gene rearrangements. The patients were treated according to the German multicenter trial of ALL in adults 05/99.5 High molecular weight DNA was prepared from bone marrow samples obtained at initial diagnosis using standard techniques. IGK and TCRD gene rearrangements were identified and characterized by PCR and sequencing as described.67 Oligonucleotides corresponding to the clone-specific junctional regions of the respective IGK/TCRD genes are shown in Table 1.

Table 1  IGK and TCRD rearrangements analyzed by dot blot hybridizations
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References

  1. Cave H, van der Werff ten Bosch J, Suciu S, Guidal C, Waterkeyn C, Otten J, Bakkus M, Thielemans K, Grandchamp B, Vilmer E . Clinical significance of minimal residual disease in childhood acute lymphoblastic leukemia New Engl J Med 1998 339: 591–598

    Article  CAS  Google Scholar 

  2. Coustan-Smith E, Behm FG, Sanchez J, Boyett JM, Hancock ML, Raimondi SC, Rubnitz JE, Rivera GK, Sandlund JT, Pui CH, Campana D . Immunological detection of minimal residual disease in children with acute lymphoblastic leukaemia Lancet 1998 351: 550–554

    Article  CAS  Google Scholar 

  3. Van Dongen JJM, Seriu T, Panzer-Grümayer ER, Biondi A, Pongers-Willemse MJ, Corral L, Stolz F, Schrappe M, Masera G, Kamps WA, Gadner H, Van Wering ER, Ludwig W-D, Basso G, de Bruijn MAC, Cazzaniga G, Hettinger K, Van der Does-van den Berg A, Hop WCJ, 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 

  4. Bartram CR . Detection of minimal residual leukemia by the polymerase chain reaction: potential implication for therapy Clin Chim Acta 1993 217: 75–83

    Article  CAS  Google Scholar 

  5. Hoelzer D, Thiel E, Löffler H, Büchner T, Ganser A, Heil G, Koch P, Freund M, Diedrich H . Prognostic factors in a multicenter study for treatment of acute lymphoblastic leukemia in adults Blood 1988 71: 123–131

    CAS  PubMed  Google Scholar 

  6. Pongers-Willemse MJ, Seriu T, Stolz F, d'Aniello E, Gameiro P, Pisa P, Gonzales M, Bartram CR, Panzer-Grümayer ER, Biondi A, San Miguel JF, van Dongen JJM . 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 Leukemia 1999 13: 110–118

    Article  CAS  Google Scholar 

  7. Nakao M, Janssen JWG, Bartram CR . Duplex PCR facilitates the identification of immunoglobulin kappa (IGK) gene rearrangements in acute lymphoblastic leukemia Leukemia 2000 14: 218–219

    Article  CAS  Google Scholar 

  8. Brown A, Akinsanya AA, Barker SJ, Brophy M, Dobb AK, Doyle SM, Hudson IR, Minter SJ, Wraith MJ, Oultram JD . Automated system for capture and detection of nucleic acids BioTechniques 1999 27: 176–180

    Article  CAS  Google Scholar 

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  1. Institute of Human Genetics, University of Heidelberg, Germany

    M Nakao, JWG Janssen, M Schäfer & CR Bartram

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  1. M Nakao
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  2. JWG Janssen
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Nakao, M., Janssen, J., Schäfer, M. et al. Digoxigenin-end labeled oligonucleotides facilitate the detection of minimal residual disease in acute lymphoblastic leukemia. Leukemia 14, 767–768 (2000). https://doi.org/10.1038/sj.leu.2401732

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