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Quantification of donor and recipient hemopoietic cells by real-time PCR of single nucleotide polymorphisms

Leukemia volume 17, pages 621–629 (2003)Cite this article

A Corrigendum to this article was published on 24 February 2004

Abstract

Analysis of changes in recipient and donor hemopoietic cell origin is extremely useful to monitor the effect of stem cell transplantation (SCT) and sequential adoptive immunotherapy by donor lymphocyte infusions (DLI). We developed a sensitive and accurate method to quantify the percentage of recipient and donor cells by real-time PCR using single nucleotide polymorphisms (SNPs) as markers. Allele-specific PCR of seven SNPs resulted in specific markers for donor or recipient in 97% of HLA-identical sibling pairs. Both, recipient- and donor-derived hemopoietic cells can be simultaneously analyzed in 67% sibling pairs. We expect this can be increased to approximately 99% by developing three additional SNP-PCR. Serial dilution of SNP-positive DNA into either SNP-negative DNA or water revealed a detection limit of 0.1–0.01% depending on the amount of input DNA and start Ct of the used SNP-PCR. Application of our real-time SNP-PCR method for a CML patient treated by allogeneic SCT and DLI demonstrated its feasibility to follow donor T-cell chimerism and early detection of residual and recurrent autologous hemopoiesis in response to treatment. This detailed monitoring of the genetic origin of hemopoietic cells, in particular immune effector cells and target cells after SCT and DLI, may substantially contribute to understanding of the mechanisms that play a role in the success of treatment.

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Acknowledgements

We are indebted to Marieke Overdijk, Paulien Polderman, Adrian van der Heijden, and Rob Woestenenk for screening recipient and donor DNA for SNPs. We thank Dr Joop Jansen for critically reading the manuscript. This work was supported by the Dutch Cancer Foundation Grant KUN 96-1363.

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  1. N Schaap and S Kolen: NS and SK contributed equally to this manuscript

Authors and Affiliations

  1. Central Hematology Laboratory, University Medical Center St Radboud, Nijmegen, The Netherlands

    F Maas, S Kolen, A Zoetbrood, H Dolstra & E van de Wiel-van Kemenade

  2. Department of Hematology, University Medical Center St Radboud, Nijmegen, The Netherlands

    N Schaap, T de Witte & A Schattenberg

  3. Bone Marrow Transplantation Unit, Hosp. G.U. Gregorio Marañon, Madrid, Spain

    I Buño

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Maas, F., Schaap, N., Kolen, S. et al. Quantification of donor and recipient hemopoietic cells by real-time PCR of single nucleotide polymorphisms. Leukemia 17, 621–629 (2003). https://doi.org/10.1038/sj.leu.2402856

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