Journal of Clinical Immunology

, Volume 38, Issue 3, pp 300–306 | Cite as

Droplet Digital PCR-Based Chimerism Analysis for Primary Immunodeficiency Diseases

  • Tsubasa Okano
  • Yuki Tsujita
  • Hirokazu KaneganeEmail author
  • Kanako Mitsui-Sekinaka
  • Kay Tanita
  • Satoshi Miyamoto
  • Tzu-Wen Yeh
  • Motoi Yamashita
  • Naomi Terada
  • Yumi Ogura
  • Masatoshi Takagi
  • Kohsuke Imai
  • Shigeaki Nonoyama
  • Tomohiro Morio
Original Article



In the current study, we aimed to accurately evaluate donor/recipient or male/female chimerism in samples from patients who underwent hematopoietic stem cell transplantation (HSCT).


We designed the droplet digital polymerase chain reaction (ddPCR) for SRY and RPP30 to detect the male/female chimerism. We also developed mutation-specific ddPCR for four primary immunodeficiency diseases.


The accuracy of the male/female chimerism analysis using ddPCR was confirmed by comparing the results with those of conventional methods (fluorescence in situ hybridization and short tandem repeat-PCR) and evaluating dilution assays. In particular, we found that this method was useful for analyzing small samples. Thus, this method could be used with patient samples, especially to sorted leukocyte subpopulations, during the early post-transplant period. Four mutation-specific ddPCR accurately detected post-transplant chimerism.


ddPCR-based male/female chimerism analysis and mutation-specific ddPCR were useful for all HSCT, and these simple methods contribute to following the post-transplant chimerism, especially in disease-specific small leukocyte fractions.


Droplet digital PCR chimerism severe combined immunodeficiency hematopoietic stem cell transplantation 



Activated PI3K-delta syndrome type 1


Confidence intervals


Droplet digital PCR


Fluorescence in situ hybridization


Graft versus host diseases


Hematopoietic stem cell transplantation


Immune dysregulation, polyendocrinopathy, enteropathy, and X-linked




Primary immunodeficiency diseases


Polymerase chain reaction


Quantitative PCR


Reduced intensity conditioning


Severe combined immunodeficiency


Short tandem repeat


Variable number tandem repeat


Wiskott-Aldrich syndrome


X-linked lymphoproliferative syndrome type 1



We thank the patients and their parents as well as the doctors who provided the samples.

Author Contributions

T.O. and Y.T. performed the experiments and wrote the manuscript. H.K. designed the study and wrote the manuscript. K.M-S., K.T., S.M., T-W.Y., M.Y., N.T., and Y.O performed the experiments. M.T., K.I., S.N., and T.M. provided critical discussion.

Funding Information

This study was supported by grants from the Ministry of Education, Culture, Sports, Science, and Technology of Japan and the Ministry of Health, Labor, and Welfare of Japan.

Compliance with Ethical Standards

We obtained written informed consent from the parents of patients and healthy controls. The study was conducted in accordance with the Declaration of Helsinki and approved by the ethics boards of Tokyo Medical and Dental University and National Defense Medical College.

Conflict of Interest

The authors declare that they have no conflict of interest.


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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  • Tsubasa Okano
    • 1
  • Yuki Tsujita
    • 2
  • Hirokazu Kanegane
    • 1
    Email author
  • Kanako Mitsui-Sekinaka
    • 2
  • Kay Tanita
    • 1
  • Satoshi Miyamoto
    • 1
  • Tzu-Wen Yeh
    • 1
  • Motoi Yamashita
    • 1
  • Naomi Terada
    • 1
  • Yumi Ogura
    • 2
  • Masatoshi Takagi
    • 3
  • Kohsuke Imai
    • 3
  • Shigeaki Nonoyama
    • 2
  • Tomohiro Morio
    • 1
  1. 1.Department of Pediatrics and Developmental Biology, Graduate School of Medical and Dental SciencesTokyo Medical and Dental UniversityTokyoJapan
  2. 2.Department of PediatricsNational Defense Medical CollegeSaitamaJapan
  3. 3.Department of Community Pediatrics, Perinatal and Maternal Medicine, Graduate School of Medical and Dental SciencesTokyo Medical and Dental University (TMDU)TokyoJapan

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