ABO chimerism determined by real-time polymerase chain reaction analysis after ABO-incompatible haematopoietic stem cell transplantation.
BACKGROUND
The dynamic alteration of ABO blood group in ABO-incompatible haematopoietic stem cell transplantation can be well defined by ABO chimerism analysis. In view of the intrinsic difference in ABO phenotypes or genotypes between donor and recipient in ABO incompatible transplantation, ABO allele-associated nucleotide polymorphic sites could theoretically be used as available informative markers for chimerism analysis.


METHODS
We chose nucleotide polymorphism sites (261, 467, 802, 803 and 1,061) of common ABO alleles to use as markers from 76 randomly chosen donors and assessed the limit of linear detection of a specifically designed real-time quantitative polymerase chain reaction using SYBR Green I dye with these sites for a chimerism assay.


RESULTS
We successfully established the real-time quantitative polymerase chain reaction for detecting 261, 467(mutated) and 803 sites and their limits of linear detection, which were at least 10(-3), 10(-2) and 10(-2), respectively. The limits of linear detection between heterozygous DNA and homozygous DNA with 261 or 803 sites were not significantly different.


CONCLUSIONS
ABO chimerism can be well analysed by real-time polymerase chain reaction with ABO gene-associated nucleotide polymorphic sites. This strategy could be very beneficial for the early and accurate judgement of the crucial point of transition in order to plan a safe transfusion strategy following ABO-incompatible transplantation.
Department of Blood Transfusion, Union Hospital of Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei Province, People's Republic of China.
Feng
F
Liu
Guining
G
Li
Xiaolu
X
Mao
Lihua
L
Hu
D000017:ABO Blood-Group System / Q000235:genetics*; D000483:Alleles; D001788:Blood Grouping and Crossmatching / Q000379:methods*; D005260:Female; D005838:Genotype; D018380:Hematopoietic Stem Cell Transplantation; D006801:Humans; D008297:Male; D010641:Phenotype; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D018183:Transplantation Chimera / Q000097:blood / Q000235:genetics*; D014184:Transplantation, Homologous
D000017:ABO Blood-Group System