Genotyping of Duffy blood group by real-time PCR with TaqMan-MGB probes
ZHOU Shihang;LIU Ming;YU Weijian;LIANG Xiaohua;Dalian Blood Station;Biological Laboratory,Dalian Medical College;
Objective To develop a new method for Duffy blood group genotyping using real-time PCR with Taq Man minor groove binding( MGB) probes and to explore the distribution of Duffy blood group alleles in Dalian Han population. Methods Primers and Taq Man-MGB probes were designed and synthesized to genotype Duffy blood group. Using the real-time PCR with Taq Man-MGB probes,a total of 120 samples were genotyped and the reproducibility and detection limit of the method were also assessed. Moreover,the real-time PCR with Taq Man-MGB probes for Duffy blood group genotyping was compared with the conventional polymerase chain reaction with allele-specific primers( PCR-ASP). Results There was a complete concordance of results for all samples genotyped by real-time PCR with Taq Man-MGB probes and PCR-ASP. The retesting results of real-time PCR with Taq Man-MGB probes were consistent with those of the initial testing. The detection limit of real-time PCR with Taq Man-MGB probes was determined as 100 pg.The FY*A and FY*B allelic frequencies were 93. 3% and 6. 7% respectively in the Chinese Han population in Dalian. The distributions of Duffy blood group in Dalian Han population did not differ significantly from that in some Chinese Han population( P 0. 05),but did differ significantly from that in Chinese She population( P 0. 05). The mismatch probability of Fyaand Fybin random transfusion was 0. 1167. Conclusion The real-time PCR with Taq Man-MGB probes for genotyping of Duffy blood group is simple,rapid,accurate,reliable,reproducible,sensitive,and superior to PCR-ASP used in routine genotyping.
【CateGory Index】： R440