Droplet Digital PCR-Based Chimerism Analysis for Primary Immunodeficiency Diseases.
OBJECTIVE
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).


METHODS
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.


RESULTS
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.


CONCLUSION
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.
['Okano|Tsubasa|T|', 'Tsujita|Yuki|Y|', 'Kanegane|Hirokazu|H|', 'Mitsui-Sekinaka|Kanako|K|', 'Tanita|Kay|K|', 'Miyamoto|Satoshi|S|', 'Yeh|Tzu-Wen|TW|', 'Yamashita|Motoi|M|', 'Terada|Naomi|N|', 'Ogura|Yumi|Y|', 'Takagi|Masatoshi|M|', 'Imai|Kohsuke|K|', 'Nonoyama|Shigeaki|S|', 'Morio|Tomohiro|T|']
D000483:Alleles D046528:Chimerism D005260:Female D018380:Hematopoietic Stem Cell Transplantation D006801:Humans D007153:Immunologic Deficiency Syndromes D017404:In Situ Hybridization, Fluorescence D008297:Male D018598:Minisatellite Repeats D009154:Mutation D060888:Real-Time Polymerase Chain Reaction D018183:Transplantation Chimera D014184:Transplantation, Homologous
chimerism hematopoietic stem cell transplantation Droplet digital PCR severe combined immunodeficiency