Detection of nucleic acids and proteins
The invention generally relates to methods for detecting a target nucleic acid and a target protein in a single assay.
1. A method of detecting multiple analytes in a sample, the method comprising the steps of:
obtaining a sample comprising at least one target nucleic acid and at least one target protein;
introducing an aptamer that binds to the target protein in the sample; and
conducting a single assay, wherein the assay detects both the target nucleic acid and the target protein, the assay comprising: removing unbound aptamer-protein complex from bound aptamer-protein complex performing PCR on the target nucleic acid; performing PCR on the aptamer protein complex; introducing a first detectably labeled probe that binds to the amplified target nucleic acid; introducing a second detectably labeled probe that binds to the amplified aptamer protein complex; detecting the first probe thereby detecting the nucleic acid in the sample; and detecting the second probe that is bound to the amplified aptamer protein complex thereby detecting the protein in the sample.
2. The method according to claim 1 , further comprising quantifying the target nucleic acid.
3. The method according to claim 1 , further comprising quantifying the target protein.
4. The method according to claim 1 , further comprising quantifying the target nucleic acid and the target protein.
5. The method according to claim 4 , wherein the quantifying comprises:
performing real time PCR on the nucleic acid; and
performing real time PCR on the aptamer, wherein quantifying the amplified aptamer quantifies the target protein.
6. The method according to claim 1 , wherein the target nucleic acid is associated with a disease.
7. The method according to claim 6 , wherein the disease is cancer.
8. The method according to claim 6 , wherein the target nucleic acid is selected from the group consisting of: FGFR3, K-ras, K-ras2, APC, DCC, TP53, PRC1, NUSAPI1, CAPZ, PFKP, EVER1, FLT1, ESPL1, AKAP2, CDC45L, RAMP, SYNGR2, NDRG1, ZNF533, and hypermethylated nucleic acid.
9. The method according to claim 1 , wherein the target protein is associated with a disease.
10. The method according to claim 9 , wherein the disease is cancer.
11. The method according to claim 9 , wherein the target protein is selected from the group consisting of: MMP-2, MMP-9, MMP9/NGAL complex, MMP/TIMP complex, MMP/TIMP1 complex, ADAMTS-7, ADAM-12, thymosin 15, thymosin 16, profilin, prohibitin, ubiquitin, tropomyosin, Cyr61, cystatin B, and chaperonin 10.
12. A method of detecting multiple analytes in a sample, the method comprising the steps of:
obtaining a sample comprising at least one target nucleic acid and at least one target protein;
introducing to the sample an aptamer that binds the target protein; and
conducting a single assay that detects and quantifies the target nucleic acid and the target protein, wherein the assay comprises removing unbound aptamer-protein complex from bound aptamer-protein complex: performing real time PCR on the nucleic acid, thereby quantifiably detecting the nucleic acid in the sample; and performing real time PCR on the aptamer-protein complex, thereby quantifiably detecting the aptamer-protein complex which quantifiably detects the target protein in the sample.
13. The method according to claim 12 , wherein the target nucleic acid is associated with a disease.
14. The method according to claim 13 , wherein the disease is cancer.
15. The method according to claim 13 , wherein the target nucleic acid is selected from the group consisting of: FGFR3, K-ras, K-ras2, APC, DCC, TP53, PRC1, NUSAPI1, CAPZ, PFKP, EVER1, FLT1, ESPL1, AKAP2, CDC45L, RAMP, SYNGR2, NDRG1, ZNF533, and hypermethylated nucleic acid.
16. The method according to claim 12 , wherein the target protein is associated with a disease.
17. The method according to claim 16 , wherein the disease is cancer.
18. The method according to claim 16 , wherein the target protein is selected from the group consisting of: MMP-2, MMP-9, MMP9/NGAL complex, MMP/TIMP complex, MMP/TIMP1 complex, ADAMTS-7, ADAM-12, thymosin 15, thymosin 16, profilin, prohibitin, ubiquitin, tropomyosin, Cyr61, cystatin B, and chaperonin 10.