摘要
目的应用Taq Man-MGB探针实时PCR技术,建立1个Duffy血型基因分型的新方法,了解大连地区汉族人群Duffy血型等位基因频率分布特征。方法设计并合成Taq Man-MGB探针实时PCR的引物和探针,应用TaqMan-MGB探针实时PCR对120例健康献血者的Duffy血型进行基因分型,对其重复性和最低检出限进行评价。并将该法和传统的等位基因特异性引物PCR(PCR-ASP)方法进行比较。结果 Taq Man-MGB探针实时PCR方法和PCR-ASP法对Duffy血型的基因分型结果是完全一致的。Taq Man-MGB探针实时PCR再次分型结果和初次基因分型的结果是完全一致的。Taq Man-MGB探针实时PCR的最低检测限是100 pg。大连汉族人群Duffy血型FY*A基因频率为93.3%,FY*B的基因频率为6.7%,其基因分布特点和国内其他地区汉族人群相似,无显著性差异(P>0.05),但与浙江畲族相比,有显著性差异(P<0.05)。Fya和Fyb抗原不配合几率为0.116 7。结论应用Taq ManMGB探针实时PCR技术建立的Duffy血型基因分型方法具有快速、简单、准确、灵敏、重复性好、通量高等特点,优于传统的PCR-ASP法。
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.
出处
《中国输血杂志》
CAS
北大核心
2016年第1期57-61,共5页
Chinese Journal of Blood Transfusion
