Method for detecting base mutation

A method for detecting nucleotide mutation is provided, thereby verifying incorrect analysis of nucleotide mutation, simultaneously detecting several nucleotide mutations adjacent to 32 nucleotides, and discriminating the presentation of nucleotide mutations in one genotype or two or more genotypes. A method for detecting nucleotide mutation comprises the steps of: (a) amplifying a specific polynucleotide using forward primer and reverse primer; (b) digesting the amplified polynucleotide with restriction enzymes form two or more single strand polynucleotide fragments having 2 to 32 nucleotides and mutation sequences; and (c) determining the molecular weight of the polynucleotide fragments, wherein the forward primer comprises a primer binding sequence 1, a restriction enzyme recognition sequence, and a primer binding sequence 2; the forward primer has the nucleotide sequence selected from the SEQ ID NOs: 2, 7, 12, 20, 25 and 30; and the restriction enzymes consist of an enzyme having low optimal temperature selected from Fok1, BbvI, BsgI, BcgI, BpmI, BseRI and BaeI, and an enzyme having high optimal temperature selected from BstF5I, TaqI, BsaBI, BrtI, BstAPI, FauI, BclI, PciI and ApoI.