Single-stranded nucleic acid for real-time detection of genetic variation of single target gene and detection method using the same

A method for simultaneously and real-time detecting six (6) different genotypes of ApoE gene, the method comprising the steps of:

• (a) obtaining a target nucleic acid comprising ApoE gene single nucleotide polymorphism sites to be detected from a biological sample;
• (b) preparing labeled single-stranded nucleic acids of SEQ ID Nos:
• 11 to 14 acting as a primer and a probe, each single-stranded nucleic acid consisting of:
• (i) a RNA region;
• (ii) a first DNA region at the 5′ side of the RNA region;
• and (iii) a second DNA region at the 3′ side of the RNA region;
• (c) mixing the target nucleic acid obtained in step (a), the single-stranded nucleic acids of SEQ ID Nos:
• 11 to 14 prepared in step (b) and a cleavage reagent, followed by amplifying the target nucleic acid through an extension reaction, wherein the single-stranded nucleic acids of SEQ ID Nos:
• 11 to 14 hybridize to the target nucleic acid at specific single nucleotide polymorphism sites of the ApoE gene and the cleavage reagent specifically cleaves the first RNA region from the first DNA region in each single-stranded nucleic acid;
• and (d) measuring the amount of the cleaved first DNA region, thereby detecting six (6) different genotypes of the ApoE gene.