Chimerism analysis by ABO blood group genotyping with digital droplet PCR
Takuya Naruto1, Maiko Sagisaka1, Mieko Ito1
1Clinical Research Institute, Kanagawa Children's Medical Center, Yokohama, Kanagawa, Japan.
Objective:
Chimerism analysis is an important post-transplant assessment for allogeneic hematopoietic stem cell transplant (HCT) recipients. Although various chimerism analysis techniques are already established, they are limited in terms of sensitivity, versatility, and turnaround time. Our objective was to develop a digital droplet polymerase chain reaction (ddPCR) assay for chimerism analysis using ABO gene polymorphisms as markers.
Methods:
Our new chimerism analysis method utilizes ddPCR to assess the ABO gene polymorphisms that encode the ABO blood genotype. ABO genotypes were determined in blood samples from 15 HCT recipients using the O panel (rs8176719) and B panel (rs8176746 and rs8176747).
Results:
The two panels distinguished six ABO genotypes (AA, AO, BB, BO, AB, and OO). The results of chimerism analysis using ABO genotypes with ddPCR were compatible with those of established methods, such as SRY gene analysis and the use of short tandem repeat markers via standard PCR. Our method could distinguish chimerism in 77% of donor and recipient combinations in the Japanese population.
Conclusions:
We developed a sensitive and rapid chimerism analysis method for HCT using ABO gene polymorphisms in ddPCR.
