Methods and Compositions for Detecting Target Sequences
First Claim
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1. A method for detecting a target nucleic acid, comprising:
- a) providing a sample comprising;
i) synthetic DNA amplified from target nucleic acid by loop-mediated isothermal amplification;
ii) oligonucleotides capable of forming an invasive cleavage structure with said synthetic DNA; and
iii) an agent for cleaving an invasive cleavage structure;
b) exposing said sample to conditions wherein said oligonucleotides form an invasive cleavage structure with said synthetic DNA, wherein said invasive cleavage structure is cleaved by said agent; and
c) detecting cleavage of an invasive cleavage structure by said agent, thereby detecting the presence of said target nucleic acid.
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Abstract
The present invention relates to compositions and methods for the detection and characterization of nucleic acid sequences and variations in nucleic acid sequences. The present invention relates to methods for amplifying a synthetic DNA from a target nucleic acid, forming a nucleic acid cleavage structure on the synthetic DNA, and detecting cleavage of the nucleic acid cleavage structure as an indicator of the preset of the target nucleic acid.
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Citations
20 Claims
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1. A method for detecting a target nucleic acid, comprising:
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a) providing a sample comprising; i) synthetic DNA amplified from target nucleic acid by loop-mediated isothermal amplification; ii) oligonucleotides capable of forming an invasive cleavage structure with said synthetic DNA; and iii) an agent for cleaving an invasive cleavage structure; b) exposing said sample to conditions wherein said oligonucleotides form an invasive cleavage structure with said synthetic DNA, wherein said invasive cleavage structure is cleaved by said agent; and c) detecting cleavage of an invasive cleavage structure by said agent, thereby detecting the presence of said target nucleic acid. - View Dependent Claims (2, 3, 4, 5, 6, 7)
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8. A method for detecting a target nucleic acid, comprising:
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a) providing a mixture comprising; i) a sample suspected of containing a target nucleic acid; and ii) detection reagents, said detection reagents comprising; (a) primers configured to produce amplified synthetic DNA from said target nucleic acid by loop-mediated isothermal amplification; (b) a polymerase for amplifying synthetic DNA from said target nucleic acid by loop-mediated isothermal amplification; (c) oligonucleotides capable of forming an invasive cleavage structure with said synthetic DNA; and (d) an agent for cleaving an invasive cleavage structure; b) exposing said mixture to conditions wherein, if said target nucleic acid is present, synthetic DNA is amplified from said target nucleic acid using said primers and said polymerase, and wherein said oligonucleotides form an invasive cleavage structure with said synthetic DNA, and wherein said invasive cleavage structure is cleaved by said agent; and c) detecting the cleavage of an invasive cleavage structure by said agent, thereby detecting the presence of said target nucleic acid. - View Dependent Claims (9, 10, 11, 12, 13, 14)
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- 15. A nucleic acid treatment kit comprising primers configured to produce amplified synthetic DNA from a target nucleic acid by loop-mediated isothermal amplification, and oligonucleotides capable of forming an invasive cleavage structure with said synthetic DNA.
Specification