Primer group and method for detecting single-base mutations

A method for detecting a nucleotide at an expected single-base mutation site of a nucleic acid sequence, the method comprising:

• providing a sample containing a nucleic acid sequence to be detected;
• amplifying the nucleic acid sequence in the sample by a polymerase chain reaction using a primer group, wherein the primer group comprises the following primers:
• an identification primer, which is composed of, from the 5′ end to the 3′ end, (a) a nucleotide sequence which complements a segment of continuous nucleotides in the nucleic acid sequence to be detected, wherein the 5′ end of the continuous nucleotides starts at a first nucleotide downstream of an expected mutation site, and (b) a nucleotide which complements a nucleotide expected to exist at the expected single-base mutation site of the nucleic acid sequence to be detected, and
• an amplification primer capable of amplifying an amplification product obtained by amplifying the nucleic acid sequence to be detected using the identification primer,
• wherein the identification primer is 1 to 19 nucleotides less than the amplification primer;
• and
• detecting whether there is a specific amplification product in a reaction product, wherein the presence of the specific amplification product indicates that there is the nucleotide expected to exist at the expected mutation site of the nucleic acid sequence to be detected.

A primer group for detecting a single-base mutation in a nucleic acid sequence, the primer group comprising the following primers:

• an identification primer, which is composed of, from the 5′ end to the 3′ end, (a) a nucleotide sequence which complements a segment of continuous nucleotides in a nucleic acid sequence to be detected, wherein the 5′ end of the continuous nucleotides starts at a first nucleotide downstream of an expected mutation site;
• and (b) a nucleotide which complements a non-mutated nucleotide or an expected mutated nucleotide at an expected single-base mutation site of a nucleic acid sequence to be detected;
• an amplification primer capable of amplifying an amplification product obtained by amplifying the nucleic acid sequence to be detected using the identification primer,
• wherein the identification primer is 1 to 19 nucleotides less than the amplification primer.