多重引物等位特异PCR法快速检测ApoE基因型的研究

Rapid Determination of Apolipoprotein E Genotypes by Multi-ASP-PCR

目的建立多重引物等位特异PCR方法结合内参照基因对ApoE基因型进行分析,并与PCR-RFLP法和直接测序结果进行比较.方法以基因组DNA为模板,用多重引物等位特异PCR法、PCR-RFLP检测了158例体检者的APOE基因型,典型基因型用直接测序法检测并通过GeneBank对比.结果共检测出5种常见的基因型:ε2/2(0%),ε3/3(77.2%),ε4/4(0.6%),ε3/2(13.3%),ε4/3(7.0%),ε4/2(1.9%),两种方法结果一致,测序结果与GeneBank对比一致.结论多重ASP-PCR检测ApoE基因型具有快速简便、准确可靠和经济、适用的特点,有一定的推广价值.

Objective To establish a method of multi allele-specific primers PCR （Muhi-ASP-PCR） with internal control gene to analyze APOE genotypes, and to compare the results with those by PCR-RFLP and those by direct nucleotide sequencing. Method APOE genotypes of 158 samples were analyzed by Muhi-ASP-PCR and PCR-RFLP. Typical genotypes were identified by direct nucleotide sequencing and the results were compared with those of Gene Bank. Results 5 common genotypes of APOE had been identified, ε3/3 （77.2%）, ε4/4 （0.6%）, ε3/2 （13.3%）, ε4/3 （7.0%）, ε4/2 （1.9%）, and forallsamples, the resuhs of Muhi-ASP-PCR were in 100% concordance with those of PCR-RFLP. The results of sequencing were in agreement with those of Gene Bank. Conclusion Muhi-ASP-PCR is a simple, rapid, reliable, economic and applicable method, which can be applied to clinical research.