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Reliable quantification of hematopoietic chimerism after allogeneic transplantation for acute leukemia using amplification by real-time PCR of null alleles and insertion/deletion polymorphisms

Leukemia volume 19, pages 336–343 (2005)Cite this article

Abstract

Increasing mixed chimerism (MC) after allogeneic stem cell transplantation (SCT) has been associated with a high risk of relapse in acute leukemia. We evaluated a new method for chimerism detection, based on the quantitative real-time PCR (qrt-PCR) amplification of null alleles or insertion/deletion polymorphisms (indels). All qrt-PCR assays with null alleles and indels attained a sensitivity of at least 10−4, as well as good intra- and interassay concordance, and a high accuracy in experiments with cell mixtures. Informativeness was found in 80.3% of the 61 donor/recipient pairs tested. Nonrelapsed patients showed a progressive decrease in peripheral blood chimerism to values below 0.01% (complete chimerism (CC)). Bone marrow chimerism failed to reach CC more than 4 years after SCT. Increasing MC was observed prior to relapse in 88.2% of patients. Compared with conventional PCR amplification of variable number of tandem repeats, qrt-PCR predicted a significantly higher number of relapses (88.2 vs 44.4%) with a median anticipation period of 58 days. In conclusion, chimerism determination by qrt-PCR amplification of null alleles and indels constitutes a useful tool for the follow-up of patients with acute leukemia after SCT, showing better results than those obtained with conventional PCR.

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Acknowledgements

We would like to thank the nursing and clinical staff, the medical residents of the Hematology Department, and the Transplantation Unit for taking care of our patients and helping us to collect the present data. We are grateful to Beli Torroba for her useful technical assistance and to Ian Johnstone for help with the English editing of the manuscript. This work was supported by grants 02/1299 and 03/0141 from the Fondo de Investigaciones Sanitarias (FIS) and grants 143/03 and 144/03 from Junta de Andalucía. GN holds fellowship Cajamar-Fundación Hospital Carlos Haya and MB FIS 01/F018

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Authors and Affiliations

  1. Hematology Department, Carlos Haya Hospital, Málaga, Spain

    A Jiménez-Velasco, M Barrios, G Navarro, A I Rodríguez, G García-Gemar & A I Heiniger

  2. Hematology Department, Reina Sofía Hospital, Córdoba, Spain

    J Román-Gómez, J A Castillejo & A Torres

  3. Bone Marrow Transplantation Unit, Gregorio Marañón Hospital, Madrid, Spain

    I Buño

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  1. A Jiménez-Velasco
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Correspondence to A Jiménez-Velasco.

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Jiménez-Velasco, A., Barrios, M., Román-Gómez, J. et al. Reliable quantification of hematopoietic chimerism after allogeneic transplantation for acute leukemia using amplification by real-time PCR of null alleles and insertion/deletion polymorphisms. Leukemia 19, 336–343 (2005). https://doi.org/10.1038/sj.leu.2403622

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