应用等位基因特异性PCR检测胆固醇酯转运蛋白基因突变

目的:建立检测胆固醇酯转运蛋白(cholesterol ester transfer protein,CETP)基因6种常见突变的等位基因特异性PCR技术。方法:应用针对CETP基因TaqIB(G→A)、I405V(A→G)、D442G(A→G)、R451Q(G→A)、A373P(G→C)和I14A(G→A)这6种常见的突变位点设计的等位基因特异性PCR技术,对海南汉、黎族人群中CETP基因突变类型进行了检测,同时对经上述等位基因特异性PCR检测的样本进行序列测定。结果:在海南汉、黎族人群中,TaqIB(G→A)突变位点可检测出GG、GA、AA3种基因型,I405V(A→G)突变位点可检测出AA、AG、GG3种基因型,D442G(A→G)突变位点可检测出AA、AG2种基因型,但在海南汉、黎族人群中未检测到R451Q(G→A)、A373P(G→C)和I14A(G→A)3种突变类型,用等位基因特异性PCR鉴定的CETP基因突变的基因分型结果与序列测定结果完全符合。结论:等位基因特异性PCR技术操作简便,重复性和稳定性好,可作为鉴定CETP基因突变类型的可行方法。

Detection of the CETP gene mutation by AS-PCR

Objective:To establish the allele specific polymerase chain reaction(AS-PCR)technique for the detection of 6 normal mutations of cholesterol ester transfer protein(CETP)gene.Methods:Designed the AS-PCR system according to the 6 mutations of CETP gene,TaqIB(G→A),I405V(A→G),D442G(A→G),R451Q(G→A),A373P(G→C)and I14A(G→A).Detected the mutations in Li and Han nationalities of Hainan province by the designed method and sequenced partial mutation samples.Results:3 genotypes were successfully detected in TaqIB(G→A)mutation site which were GG,GA,AA,3 genotypes were successfully detected in I405V(A→G)mutation site which were AA,AG and GG,and 2 genotypes were successfully detected in D442G(A→G)mutation site which were AA and AG,while only the wild genotype could be detected separately in R451Q(G→A),A373P(G→C)and I14A(G→A)mutation sites which was GG.All the results above provided by the AS-PCR were exactly consistent with the sequencing results.Conclusion:The AS-PCR with advantages of stability and simplify is an effective method for investigation of the mutations of CETP gene.