Molecular detection using ligation amplification

Abstract

Disclosed herein are compositions, kits, methods, and systems for detecting a target molecule in a sample using a detection molecule.

1 - 177 . (canceled) 
     
     
         178 . A method for amplifying a signal of a target molecule in an assay, comprising:
 a) contacting the target molecule in the assay with a detection molecule and an antisense oligomer, wherein the detection molecule comprises: i) a ligand that binds the target molecule; and ii) a single-stranded nucleic acid linked to the ligand, and wherein the single-stranded nucleic acid is capable of hybridizing to the antisense oligomer to form a double-stranded nucleic acid in situ, and   b) adding to the assay two or more detectors, wherein a first detector is configured for ligating with the double-stranded nucleic acid, and a second detector is configured for ligating in situ with the first detector to amplify the signal of the target molecule.   
     
     
         179 . The method of  claim 178 , wherein the assay is a western blot assay, an enzyme-linked immunosorbent assay (ELISA), a fluorescent microbead based assay, or any combination thereof. 
     
     
         180 . The method of  claim 178 , wherein the two or more detectors are added sequentially. 
     
     
         181 . The method of  claim 178 , wherein the two or more detectors comprises a third detector, and wherein the third detector is configured for ligating in situ with the first detector and/or the second detector, to further amplify the signal of the target molecule. 
     
     
         182 . The method of  claim 181 , wherein the two or more detectors comprises one or more additional detectors that is configured for ligating in situ with the first detector, the second detector, and/or the third detector, to further amplify the signal of the target molecule. 
     
     
         183 . The method of  claim 178 , wherein at least one of the two or more detectors comprises a detection tag. 
     
     
         184 . The method of  claim 183 , wherein the detection tag comprises a fluorophore. 
     
     
         185 . The method of  claim 184 , wherein the fluorophore comprises coumarin, rhodamine, xanthene, fluorescein, or cyanine. 
     
     
         186 . The method of  claim 183 , wherein the detection tag comprises a hapten. 
     
     
         187 . The method of  claim 186 , wherein the hapten comprises a biotin, digoxigenin, fluorescein, aniline, urushiol, hydralazine, or dinitrophenol. 
     
     
         188 . The method of  claim 183 , wherein the detection tag comprises an enzyme. 
     
     
         189 . The method of  claim 188 , wherein the enzyme comprises an alkaline phosphatase or HRP. 
     
     
         190 . The method of  claim 178 , wherein target molecule comprises an antigen. 
     
     
         191 . The method of  claim 178 , wherein target molecule comprises a primary antibody. 
     
     
         192 . The method of  claim 178 , wherein target molecule comprises a secondary antibody. 
     
     
         193 . The method of  claim 178 , wherein the target molecule comprises a cell surface molecule. 
     
     
         194 . The method of  claim 178 , wherein the target molecule comprises a nucleic acid, a carbohydrate, or a lipid. 
     
     
         195 . The method of  claim 178 , wherein the target molecule comprises a protein. 
     
     
         196 . The method of  claim 195 , wherein the protein comprises an immune system protein, a cytoskeletal protein, an extracellular matrix protein, a plasma protein, a coagulation factor, an acute phase protein, a hemoprotein, a cell adhesion, a transmembrane transport protein, an ion channel, a symport/antiport protein, a hormone, a growth factor, a receptor, a DNA-binding protein, a RNA-binding protein, a transcription regulatory protein, a nutrient storage or transport protein, a chaperone protein, an enzyme, or any combination thereof. 
     
     
         197 . A composition that comprises:
 a) a detection molecule and an antisense oligomer, wherein the detection molecule comprises: i) a ligand that binds a target molecule; and ii) a single-stranded nucleic acid linked to the ligand, and wherein the single-stranded nucleic acid is capable of hybridizing to the antisense oligomer to form a double-stranded nucleic acid in situ, and   b) two or more detectors, wherein a first detector is configured for ligating with the double-stranded nucleic acid, and a second detector is configured for ligating in situ with the first detector.   
     
     
         198 . The composition of  claim 197 , wherein the two or more detectors comprises a third detector, and wherein the third detector is configured for ligating in situ with the first detector and/or the second detector, to further amplify the signal of the target molecule. 
     
     
         199 . The composition of  claim 198 , wherein the two or more detectors comprises one or more additional detectors that is configured for ligating in situ with the first detector, the second detector, and/or the third detector, to further amplify the signal of the target molecule.