Abstract: There are significant individual differences in the antiplatelet effects of aspirin. Three single nucleotide polymorphisms(SNPs), rs5918, rs12041331 and rs730012, are reported to significantly correlate with the efficacy and side effects of aspirin. In the present study, the genotyping method of the three SNPs was established based on the combination of polymerase chain reaction and pyrosequencing technology. Amplification and sequencing primers were designed independently; the amplification conditions were optimized to amplify the three SNPs in the same condition. The sensitivity of the method was detected using original genome DNA at different concentrations. In order to testify the accuracy of the method, the proposed method and Sanger sequencing technology were both used to genotype the three SNPs in 20 blood samples. The results demonstrated that the genotyping method of aspirin-related SNPs was successfully established, with the detection limit being as low as 0. 4 ng genome DNA. The genotype results of 20 samples by the proposed method were exactly the same as that of Sanger sequencing. It is evident that the proposed method is sensitive and accurate.