HIGH SPECIFICITY AND HIGH SENSITIVITY DETECTION BASED ON STERIC HINDRANCE & ENZYME-RELATED SIGNAL AMPLIFICATION
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Abstract
The present invention relates to a molecular probe capable of high sensitivity and high specificity detection of a target nucleic acid in a sample. Also disclosed is a detection method using this probe.
6 Citations
25 Claims
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13. A method of assaying a target nucleic acid molecule in a sample, the method comprising:
- contacting a probe comprising a ligand and a polynucleotide sequence, wherein the polynucleotide sequence is capable of forming a hairpin and the polynucleotide sequence comprises a 5′
domain and a 3′
domain, wherein the 5′
domain comprises the first six to 10 nucleotides of the 5′
end of the polynucleotide sequence and the 5′
domain is not complementary to the target nucleic acid molecule and wherein the 3′
domain is from 27 to 41 nucleotides in length and can specifically hybridize to a sequence of the target nucleic acid molecule, and is capable of hybridizing with the 5′
domain, wherein hybridization of the 5′ and
3′
domains would produce a loop of at least 21 nucleotides and wherein the polynucleotide sequence is no longer than 51 nucleotides, the probe having a first three-dimensional structure in the absence of the target nucleic acid molecule being bound thereto and a second three-dimensional structure in the presence of the target nucleic acid being bound thereto, wherein the first three-dimensional structure inhibits or prevents the ligand from binding a receptor and the second three-dimensional structure allows the ligand to bind to the receptor., thereby forming a ligand receptor complex wherein the ligand receptor complex results in a detectable signal wherein the detectable signal increases as the number of nucleotides in the 5′
domain increases from six to 10. - View Dependent Claims (14, 15, 16, 17, 18, 19, 20, 21, 23, 24, 25)
- contacting a probe comprising a ligand and a polynucleotide sequence, wherein the polynucleotide sequence is capable of forming a hairpin and the polynucleotide sequence comprises a 5′
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Specification