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Donor-derived DNA variability in fingernails of acute myeloid leukemia patients after allogeneic hematopoietic stem cell transplantation detected by direct PCR

Bone Marrow Transplantation volume 55, pages 1021–1022 (2020)Cite this article

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Human fingernails are an important material used to identify individuals in forensic medicine as well as a source of DNA for genetic analyses in different fields of study. Nail clippings have many advantages, such as stability and suitability for long-term storage, easy non-invasive sampling and simple transport [1, 2]. On the other hand, the structure of a fingernail (presence of keratinized cells) requires a more complicated DNA extraction method than DNA extraction from blood cells. Therefore, it is advantageous to use a direct PCR method that does not require a DNA isolation step. This method was used on fingernails for the first time by Tie et al. [3], and by Ottens et al. [4]. The benefits of the method are simplicity, reducing the possibility of contamination and time- and cost-saving. We used this innovative method for donor-derived DNA analysis on the fingernails of 30 acute myeloid patients (AML) with long-lasting complete donor chimerism in their peripheral blood after the first allogeneic hematopoietic stem cell transplantation (allo-HSCT).

The nails were collected between 217 to 6169 days after the first allo-HSCT from patients that had complete donor chimerism (CC) in the peripheral blood at least 110 days before nail collection. Specific short tandem repeat (STR) polymorphisms were determined for each donor-recipient pair in the peripheral blood (before allo-HSCT) and the same STR polymorphisms were used for fingernails analysis by direct PCR. A total of 15 different STR markers were used for fingernail analysis. In 26/30 cases we were able to amplify at least two informative STR polymorphisms per patient by direct PCR. Each fingernail (at least one per patient) was divided into 3–6 fragments and each of these fragments was analyzed by direct PCR in two ways: (i) as a pooled sample (in an equivalent ratio) composed from the same patient’s partial nail fragments to analyze interindividual variability, and (ii) as separate samples of partial nail fragments to identify intraindividual variability.

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Acknowledgements

Supported by Ministry of Health, Czech Republic—conceptual development of research organization (FNBr, 65269705).

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Authors and Affiliations

  1. Department of Internal Medicine—Hematology and Oncology, University Hospital Brno, Brno, Czech Republic

    Blanka Robesova, Adam Folta, Marta Krejci, Frantisek Folber, Zuzana Sustkova, Jiri Mayer & Ivana Jeziskova

  2. Department of Animal Morphology, Physiology and Genetics, Faculty of AgriSciences, Mendel University in Brno, Brno, Czech Republic

    Marie Drncova

  3. Department of Internal Medicine—Hematology and Oncology, Faculty of Medicine, Masaryk University, Brno, Czech Republic

    Marta Krejci, Frantisek Folber, Zuzana Sustkova, Jiri Mayer & Ivana Jeziskova

  4. Central European Institute of Technology (CEITEC), Masaryk University, Brno, Czech Republic

    Jiri Mayer

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  1. Blanka Robesova
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Contributions

BR, JM, and IJ designed the study. BR, MD, and AF performed experiments. BR, AF, and IJ analyzed data. MK, FF, and ZS provided the patients samples and clinical data. BR, AF, and IJ wrote the article. MD, MK, FF, ZS, and JM revised the manuscript. All authors have read and agreed with the final version of the manuscript.

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Correspondence to Ivana Jeziskova.

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Robesova, B., Drncova, M., Folta, A. et al. Donor-derived DNA variability in fingernails of acute myeloid leukemia patients after allogeneic hematopoietic stem cell transplantation detected by direct PCR. Bone Marrow Transplant 55, 1021–1022 (2020). https://doi.org/10.1038/s41409-020-0938-x

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  • DOI: https://doi.org/10.1038/s41409-020-0938-x

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