| 主权项 |
1. A method of detecting a target nucleic acid, the method comprising:
incubating a first polynucleotide and a sample comprising a target nucleic acid, whereby the target nucleic acid and the first polynucleotide hybridize to form a hybridization product,
wherein the first polynucleotide comprises a first complementary region complementary to at least two continuous nucleotides of a first region of the target nucleic acid; and a second complementary region complementary to at least two continuous nucleotides of a second region of the target nucleic acid, wherein the second region of the target nucleic acid is located 3′ of the first region on the target nucleic acid, and the second complementary region is located 3′ of the first complementary region on the first polynucleotide; incubating the hybridization product in the presence of a first nucleic acid polymerase to extend the first polynucleotide to prepare an extended product; incubating the extended product in the presence of a nucleic acid ligase to prepare a second polynucleotide having a cyclic structure; incubating the second polynucleotide having a cyclic structure in the presence of a primer set and a second nucleic acid polymerase to amplify the second polynucleotide having a cyclic structure to prepare an amplification product,
wherein the primer set comprisesa forward primer comprising a polynucleotide sequence that is identical to at least two continuous nucleotides of a third region of the first polynucleotide located between the first complementary region and the second complementary region of the first polynucleotide, and a reverse primer comprising a polynucleotide sequence that is identical to at least two continuous nucleotides located between the first region and the second region of the target nucleic acid, ora forward primer comprising a polynucleotide sequence that is identical to at least two continuous nucleotides located between the first region of the target nucleic acid and the second region of the target nucleic acid, and a reverse primer comprising a polynucleotide sequence that is identical to at least two continuous nucleotides located between the first complementary region of the first polynucleotide and the second complementary region of the first polynucleotide; and detecting the amplification product to detect the target nucleic acid. |
