Materials and methods for rapid and specific detection of synthetic cathinones
View Patent ↗The subject invention provides methods, assays, and products for detecting small molecules in a sample, in particular, in both clinical and field settings. The method for detecting a small-molecule target, preferably, a synthetic cathinone in a sample comprises contacting the sample with an aptamer-based sensor selective for the small-molecule target, and detecting the small-molecule target in the sample. Specifically, the method utilizes an aptamer-based sensor comprising a dye binding to a three-way junction binding domain of an aptamer. Binding of small-molecule target to the aptamer displaces the dye, generating a spectroscopic signal that can be used for detection of the small-molecule target and quantitative measurement of the target concentration.
1. An aptamer-based sensor comprising an aptamer, the aptamer comprising SEQ ID NO: 12 and having a maximal length of 73 nucleotides.
2. The aptamer-based sensor according to claim 1 , the aptamer being SEQ ID NO: 12.
3. The aptamer-based sensor according to claim 1 , the aptamer having a maximal length of 53 nucleotides.
4. The aptamer-based sensor according to claim 1 , the aptamer having a maximal length of 51 nucleotides.
5. The aptamer-based sensor according to claim 1 , the aptamer comprising SEQ ID No: 13 and having a maximal length of 73 nucleotides.
6. The aptamer-based sensor according to claim 1 , the aptamer comprising SEQ ID No: 14 and having a maximal length of 73 nucleotides.
7. The aptamer-based sensor according to claim 1 , the aptamer being SEQ ID No: 13.
8. The aptamer-based sensor according to claim 1 , the aptamer being SEQ ID No: 14.
9. The aptamer-based sensor according to claim 1 , the aptamer binding to a dye.
10. The aptamer-based sensor according to claim 9 , the dye being Cy7.
11. A method for detecting a synthetic cathinone in a sample comprising contacting the sample with the aptamer-based sensor according to claim 1 , and detecting the synthetic cathinone in the sample, the detection comprising measuring a signal generated upon binding of the small-molecule target to the aptamer-based sensor, the signal being a change in absorbance or fluorescence intensity.
12. The method according to claim 11 , wherein the sample is a biological sample or an environmental sample.
13. The method according to claim 12 , wherein the biological sample is selected from blood, plasma, urine, tears, and saliva.
14. The method according to claim 11 , wherein the synthetic cathinone being 3, 4-methylenedioxypyrovalerone (MDPV), naphyrone, methylone, pentylone, mephedrone, ethylone, butylone, pyrovalerone, 3-methylmethcathinone (3-MMC), or 3′,4′-methylenedioxy-α-pyrrolidinopropiophenone (MDPPP).
15. The method according to claim 11 , the aptamer-based sensor comprising a dye.
16. The method according to claim 15 , the dye being Cy7.
17. The method according to claim 11 , the synthetic cathinone having a core structure of:
wherein R 1 and R 2 , are each independently selected from the group consisting of hydrogen, alkyl, and alkoxy; or R 1 and R 2 , taken together with the carbon atoms to which they are attached, form a substituted or unsubstituted 5- or 6-membered methylenedioxy group or aromatic ring;
R 3 is alkyl;
R 4 and R 5 are each independently selected from the group consisting of hydrogen and alkyl; or R 4 and R 5 , taken together with the nitrogen atom to which they are attached, form a substituted or unsubstituted 5- or 6-membered pyrrolidino group; and
R 6 is hydrogen.
18. The method according to claim 17 , wherein R 1 and R 2 together form a methylenedioxy group or an aromatic ring, and R 4 and R 5 together form a pyrrolidino group.