等位基因特异荧光PCR检测ALDH2基因多态性

Polymorphism of ALDH2 gene with allele specific fluorescence polymerase chain reaction

目的了解人群乙醛脱氢酶2(ALDH2)基因多态性的分布,评价等位基因特异荧光聚合酶链反应(PCR)检测ALDH2基因应用于临床的可行性。方法采用等位基因特异荧光PCR和DNA测序法同时检测375例患者外周血ALDH2基因型,结果不一致的标本采用基因芯片技术进行复检。结果 375例标本中,荧光PCR检出ALDH2*1/1型248例(66.1%)、ALDH2*1/2型107例(28.5%)、ALDH2*2/2型20例(5.4%),而DNA测序法则分别检出246例(65.6%)、108例(28.8%)和21例(5.6%),2种方法检测ALDH2基因型差异无统计学意义(χ~2=1.33,P=0.392),且一致性较好(κ=0.978,P<0.001)。2种方法基因型检测结果一致率为98.9%(371/375)。4例结果不一致的标本经基因芯片法复检,结果与等位基因特异荧光PCR一致。结论等位基因特异荧光PCR检测ALDH2基因型简便、可靠,能满足临床对ALDH2基因型检测的要求。

Objective To investigate the distribution of acetaldehyde dehydrogenase2(ALDH2)genepolymorphism,and to evaluate the feasibility of allele specific fluorescence polymerase chain reaction(PCR)for ALDH2gene polymorphism.Methods Allele specific fluorescence PCR and DNA sequencing were used for ALDH2genotyping in peripheral blood of375patients.Gene chip method was adopted to determine the specimens with inconsistent results by the2methods.Results Among the375specimens,248cases(66.1%)of ALDH2*1/1,107cases(28.5%)of ALDH2*1/2and20cases(5.4%)of ALDH2*2/2were determined by allele specificfluorescence PCR,while246cases(65.6%)of ALDH2*1/1,108cases(28.8%)of ALDH2*1/2and21cases(5.6%)of ALDH2*2/2were determined by DNA sequencing,respectively.There was no statistical significance for ALDH2genotyping between allele specific fluorescence PCR and DNA sequencing(χ2=1.33,P=0.392),with good consistency(κ=0.978,P<0.001).The coincidence rate was98.9%(371/375)between the2methods.For4cases with inconsistent results,the results of gene chip method was in agreement with those by allele specificfluorescence PCR.Conclusions Allele specific fluorescence PCR for ALDH2genotyping is feasibleand could meet the requirements of clinical ALDH2gene determination.