[Detection of hematopoietic chimera by real-time fluorescent quantitative PCR with erythrocyte Kidd blood group gene].
This study was aimed to establish the real-time fluorescent quantitative PCR (RT-qPCR) with erythrocyte Kidd blood group gene for detecting the hematopoietic chimera and to investigate the feasibility of this method. The TaqMan MGB probes and special primers were designed on basis of difference of erythrocyte Kidd blood group alleles, the hematopoietic chimerism was detected by RT-qPCR, the DNA chimerism was simulated by means of dilution of multiple proportions, and the sensitivity analysis was performed. The results showed that the RT-qPCR with erythrocyte Kidd blood group gene could effectively distinguish JK*A and JK*B alleles. There was no significant difference between the theoretic value and the practical measured value by this method (P > 0.05). As 156 donor's cells could be discriminated from 10(4) chimeric cells, this method may effectively detect donor's cells with correlation coefficient 0.998. It is concluded that the established RT-qPCR with erythrocyte Kidd blood group gene shows the feasibility for quantitative detection of hematopoietic chimera, and may be used to quantitatively detect chimera in a certain range.
Zhejiang University Medical College, Zhejiang Province, China.
Shu
S
Chen
Xian-Guo
XG
Xu
Ying
Y
Liu
Xiao-Zhen
XZ
Hong
Fa-Ming
FM
Zhu
Hang-Jun
HJ
Lü
Li-Xing
LX
Yan
D002678:Chimera; D004912:Erythrocytes; D006801:Humans; D007667:Kidd Blood-Group System / Q000235:genetics*; D060888:Real-Time Polymerase Chain Reaction
D007667:Kidd Blood-Group System