NUCLEIC ACID AMPLIFICATION METHOD USING ALLELE-SPECIFIC REACTIVE PRIMER

The present invention relates to a nucleic acid amplification method using an allele-specific reactive primer (ASRP) which is designed to solve problems of conventional allele-specific PCR. More specifically, the present invention relates to a detection method of a target nucleic acid, comprising a DNA polymerase having proofreading activity and a base sequence complementary to the target nucleic acid, wherein the target nucleic acid is amplified in the presence of an ASRP which is modified in such a manner that one or more bases from the base immediately in front, in the 5'-end direction, of a base that is cut off by the proofreading activity of the DNA polymerase when a non-complementary base exists at the 3' end, to the base at the 5' end, cannot serve as a primer for polymerization. The detection method using an ASRP according to the present invention is a technique with very high specificity of amplification due to the characteristics of the ASRP and the proofreading DNA polymerase, and is capable of effectively detecting mutations (point mutation, insertion, and deletion) including single nucleotide polymorphism (SNP). Furthermore, the method can also be used for detecting the presence of CpG methylation after bisulfite treatment, or for amplifying and detecting a target DNA starting with a desired base sequence from a DNA library.