Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Correspondence
  • Published:

Buccal swabs but not mouthwash samples can be used to obtain pretransplant DNA fingerprints from recipients of allogeneic bone marrow transplants

Bone Marrow Transplantation volume 25, pages 575–577 (2000)Cite this article

With great interest we read a recent report by Endler and coworkers.1 In this paper, the authors show that DNA derived from mouthwashes after allogeneic blood stem cell transplantation frequently displays a chimeric or complete donor genotype and is thus not suitable to obtain information on the pre-transplantation allelotype of the recipient. Although we agree with the results obtained in this study, we would like to report our experiences on buccal epithelial cells as a source for chimerism analysis.

DNA-based assessment of chimerism after allogeneic blood stem cell transplantation (BSCT) using highly polymorphic DNA sequences, such as short tandem repeat (STR) markers, has become a widely used method because of the sensitivity, the ease of performance and the independence of sex mismatch between donor and recipient. A limitation frequently imposing uncertainties in the interpretation of the results is that a pre-transplantation sample has to be analyzed for donor and recipient, in order to define informative STR combinations. Although this should not be difficult in theory, our experience shows that in 10–15% of all patients, the corresponding pre-transplantation sample is missing. The oral mucosa represents an attractive source of appropriate material, since it is easily accessible and should theoretically be derived from the recipient. However, Endler et al1 found that mouthwashes performed after BSCT frequently contain a high proportion of donor cells and sometimes the DNA derived from this material completely reflects the donor STR profile. We have employed a different approach, aiming to directly use epithelial cells derived from buccal swabs to obtain the pre-transplantation STR genotype of the recipient. This technique is standard in forensic medicine. However, after allogeneic transplantation the situation is complicated, since granulocytes and lymphocytes, which are trafficking through the normal human mucosa and can be found in the saliva, are usually derived from the donor. There is highly divergent information on the number of leukocytes in the normal saliva, ranging from 2 to 136000 cells/ml, and in patients with inflammation of the oral cavity, this number was found to be up to 1.1 × 106/ml.2 Thus, in order to remove as many non-epithelial cells as possible, patients were asked to thoroughly rinse their mouth twice with 5–10 ml of water. The first portion of rinsing fluid was used as mouthwash sample (see below). Then, a sterile cytological brush (Cytosoft Cytology Brush, Medical Packaging, Camarillo, CA, USA) was employed to collect epithelial cells from the buccal mucosa on both sides of the oral cavity. Cells were immediately recovered by immersion in 200 μl PBS. DNA was subsequently extracted from this sample and an aliquot of the mouthwash sample using a silica-based procedure (Qiagen DNA Blood Kit; Qiagen, Hilden, Germany). The DNA was eluted in 40–50 μl of TE buffer (10 mM Tris, 0.1 mM EDTA, pH 8), quantified and diluted to a final concentration of 0.125 ng/μl in TE buffer. Usually, the DNA derived from the buccal swabs was too dilute to be accurately quantified. In these cases, a 1/10 dilution of the original sample was used. Quantitative chimerism analysis was performed using a recently developed procedure based on a multiplex PCR with coamplification of nine STRs labeled with fluorescent primers.3 PCR, gel electrophoresis and calculation of the donor/recipient ratio were performed as described.

This is a preview of subscription content, access via your institution

Relevant articles

Open Access articles citing this article.

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1

References

  1. Endler G, Greinix H, Winkler K et al. Genetic fingerprinting in mouthwashes of patients after allogeneic bone marrow transplantation Bone Marrow Transplant 1999 24: 95–98

    Article  CAS  Google Scholar 

  2. Burnett GW, Scherp HW . Oral Microbiology and Infectious Disease Williams & Wilkins Co: Baltimore 1962 pp 322–323

  3. Thiede C, Florek M, Bornhäuser M et al. Rapid quantification of mixed chimerism using multiplex amplification of short tandem repeat markers and fluorescence detection Bone Marrow Transplant 1999 23: 1055–1060

    Article  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Medizinische Klinik und Poloklinik I, Universitätsklinikum Carl Gustav Carus, Technische Universität Dresden, Fetscherstr. 74, Dresden, 01307, Germany

    C Thiede, G Prange-Krex, J Freiberg-Richter, M Bornhäuser & G Ehninger

Authors
  1. C Thiede
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. G Prange-Krex
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. J Freiberg-Richter
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. M Bornhäuser
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. G Ehninger
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Thiede, C., Prange-Krex, G., Freiberg-Richter, J. et al. Buccal swabs but not mouthwash samples can be used to obtain pretransplant DNA fingerprints from recipients of allogeneic bone marrow transplants. Bone Marrow Transplant 25, 575–577 (2000). https://doi.org/10.1038/sj.bmt.1702170

Download citation

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/sj.bmt.1702170

This article is cited by

Search

Advanced search

Quick links