Sequence Conversion and Signal Amplifier DNA Having Poly DNA Spacer Sequences and Detection Methods Using Same
AssigneesAbbott Japan Co., Ltd., Tokyo Institute of Technology
InventorsKen KOMIYA, Makoto KOMORI, Toru YOSHIMURA
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Abstract
Disclosed are methods for detecting a target nucleic acid in a sample. The methods include contacting said sample, in the presence of a polymerase and an endonuclease, with a sequence conversion oligonucleotide. Also disclosed are methods for detecting a target nucleic acid in a sample in which said sample is contacted, in the presence of a polymerase and an endonuclease, with a sequence conversion oligonucleotide and a signal amplifier oligonucleotide. The disclosure also provides compositions and kits comprising such sequence conversion and signal amplifier oligonucleotides.
Claims37
A method for detecting a target nucleic acid in a sample, said method comprising contacting said sample with:
- a first oligonucleotide comprising, in the 5′ to 3′ direction, a signal DNA generation sequence, an endonuclease recognition site, a poly DNA spacer sequence, and a sequence complementary to the 3′ end of said target nucleic acid;
- a second oligonucleotide comprising, in the 5′ to 3′ direction, a signal DNA generation sequence homologous to the signal DNA generation sequence of the first oligonucleotide, an endonuclease recognition site, a poly DNA spacer sequence, and a sequence that is homologous to the signal DNA generation sequence of the first oligonucleotide;
- a polymerase;
- and
- an endonuclease for a nicking reaction.
A composition for detecting a target nucleic acid in a sample, said composition comprising:
- a first oligonucleotide comprising, in the 5′ to 3′ direction, a signal DNA generation sequence, an endonuclease recognition site, a poly DNA spacer sequence, and a sequence complementary to the 3′ end of said target nucleic acid;
- a second oligonucleotide comprising, in the 5′ to 3′ direction, a signal DNA generation sequence homologous to the signal DNA generation sequence of the first oligonucleotide, an endonuclease recognition site, a poly DNA spacer sequence, and a sequence that is homologous to the signal DNA generation sequence of the first oligonucleotide.
A kit for detecting a target nucleic acid in a sample, said kit comprising:
- a first oligonucleotide comprising, in the 5′ to 3′ direction, a signal DNA generation sequence, an endonuclease recognition site, a poly DNA spacer sequence, and a sequence complementary to the 3′ end of said target nucleic acid;
- and
- a second oligonucleotide comprising, in the 5′ to 3′ direction, a signal DNA generation sequence homologous to the signal DNA generation sequence of the first oligonucleotide, an endonuclease recognition site, a poly DNA spacer, and a sequence that is homologous to the signal DNA generation sequence of the first oligonucleotide.
The kit according to any of claims 21 to 32, further comprising instructions for use.
A chemically modified oligonucleotide comprising, in the 5′ to 3′ direction, a first sequence that is complementary to a known signal DNA sequence, an endonuclease recognition site, a poly DNA spacer sequence, and a second sequence that is complementary to the same known signal DNA sequence as the first sequence.
Dependencies are inferred from the claim text.