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Chimaerism

Analysis of haematopoietic chimaerism by quantitative real-time polymerase chain reaction

Bone Marrow Transplantation volume 35, pages 283–290 (2005)Cite this article

Summary:

Allogeneic bone marrow transplantation (BMT) with marrow ablative conditioning is the treatment of choice for haematopoietic malignancies. The use of nonmyeloablative stem cell transplants has allowed the treatment of patients previously ineligible for BMT because of age or other disease. These reduced conditioning regimes allow the persistence initially of some recipient cells in the blood and bone marrow (haematopoietic chimaerism). Monitoring of the relative proportion of donor and recipient cells is required to assess the success of the procedure, to predict subsequent rejection or impending relapse and to guide the use of donor lymphocyte infusions. We present a quantitative real-time PCR approach for the measurement of haematopoietic chimaerism using the TaqMan™. This approach exploits the presence of single-nucleotide polymorphisms (SNPs) to distinguish cells of patient or donor origin. We have designed and validated a panel of seven allele-specific probes to quantify the contribution of patient and donor cells in the haematopoietic population from 12 patient and donor pairs. We have compared the performance of this approach with an existing method and proved it to be superior in both accuracy and sensitivity. The use of more sensitive and accurate techniques permits earlier intervention for improved clinical outcome.

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Acknowledgements

This work was funded by a small research grant from the Royal Devon and Exeter NHS Foundation Trust R&D Directorate. We thank the Exeter Leukaemia Fund for funding the purchase of additional probes. We thank Dr Richard Lee, Dr Marilyn Pocock and Dr Claudius Rudin at the Royal Devon and Exeter NHS Foundation Trust for providing samples and valuable advice; Dr Tim Frayling from the Peninsula Medical School for designing the T515 primers and probes and Mr Neil Goodman for carrying out the artificial mixing experiments.

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Authors and Affiliations

  1. Department of Molecular Genetics, Royal Devon and Exeter NHS Foundation Trust, Barrack Road, Exeter, Devon, England

    L W Harries, C L Wickham, J C Evans & S Ellard

  2. Institute of Biomedical and Clinical Science, Peninsula Medical School, Exeter, UK

    L W Harries & S Ellard

  3. Department of Haematology, Derriford Hospital NHS Healthcare Trust, Plymouth, UK

    S A Rule

  4. Department of Haematology, Royal Devon and Exeter NHS Foundation Trust, Barrack Road, Exeter, Devon, England

    M V Joyner

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  1. L W Harries
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Correspondence to L W Harries.

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Harries, L., Wickham, C., Evans, J. et al. Analysis of haematopoietic chimaerism by quantitative real-time polymerase chain reaction. Bone Marrow Transplant 35, 283–290 (2005). https://doi.org/10.1038/sj.bmt.1704764

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