Single nucleotide polymorphisms (SNPs) and single-base mismatch in genome are closely associated with disease development,progress,treatment,and prognosis [1].Many methods have been developed to detect SNPs.Some assays,such as Taqman MGB probe and high resolution melting,amplify and detect both wild-type and mutant genomic DNA by specific primers and probes [2],while some other assays,such as ARMS and PNA-LNA clamp PCR,only amplify and detect mutant templates [2,3].Amplification Refractory Mutation System is widely used to detect SNP and mutations;however,the detection limit is ～ 1％ to 5 ％ of mutants in wild-type DNA content [4].