Novel method for lateral flow immunoassay

Abstract

The present disclosure relates to a novel method for lateral flow immunoassay (LFIA) by utilizing plasmonic enhancement strategy. More specifically, the present disclosure provides a plasmonic enhanced lateral flow sensor (pLFS) concept by introducing a liposome-based amplification of the colorimetric signals on the lateral flow platform for ultrasensitive detection of pathogens.

We claim: 
     
         1 . A method of detecting one or more target analytes in a liquid sample, wherein the method comprises:
 a) providing a lateral flow strip, wherein the lateral flow strip comprises a sample pad to receiving a liquid sample, a conjugate pad, a sample capture area having pre-loaded antibodies that are capable of binding a target analyte;   b) preparing a liquid sample comprising a mixture of a target analyte, streptavidin, gold nanoparticles (GNPs) probes comprising biotinylated and antibody modified gold nanoparticles (GNPs), and an amine-containing chemical loaded liposomes;   c) loading said liquid sample solution to the sample pad of the lateral flow strip and let the liquid sample migrate along the lateral flow strip to let the target analyte bind with the pre-loaded antibody on the capture area; and   d) providing a detecting agent comprising gold nanoparticles and a hydrolytic agent and allowing the detecting agent to contact the lateral flow strip one or more times to generate a colorimetric signal that shows the existence of the target analyte, wherein the hydrolytic agent is capable of hydrolysis of said liposomes to release said amine-containing chemical.   
     
     
         2 . The method of  claim 1 , wherein the target analyte comprises a pathogen. 
     
     
         3 . The method of  claim 1 , wherein the pathogen is  E. coli  O157:H7,  Salmonella  sp.,  Listeria  sp., or any combination thereof. 
     
     
         4 . The method of  claim 1 , wherein the amine-containing chemical is branched polyethylenimine, cysteine, salt or derivative therefore, or any combination thereof. 
     
     
         5 . The method of  claim 1 , wherein the target analyte can be observed by naked human eyes through colored line and/or spot corresponding to the availability of the target analyte. 
     
     
         6 . The method of  claim 1 , wherein the target analyte can be identified within one hour period. 
     
     
         7 . The method of  claim 1 , wherein the method has no enzyme being used. 
     
     
         8 . The method of  claim 1 , wherein the hydrolytic agent comprises polyoxyethylene (20) sorbitan monolaurate. 
     
     
         9 . A liquid composition comprising a mixture of streptavidin, gold nanoparticles (GNPs) probes comprising biotinylated and antibody modified gold nanoparticles (GNPs), and an amine-containing chemical loaded liposomes, wherein the composition is used to bind to a target analyte. 
     
     
         10 . The liquid composition of  claim 9 , further comprising a target analyte. 
     
     
         11 . The liquid composition of  claim 9 , wherein the target analyte comprise pathogen. 
     
     
         12 . The liquid composition of  claim 11 , wherein the pathogen is  E. coli  O157:H7,  Salmonella  sp.,  Listeria  sp., or any combination thereof. 
     
     
         13 . The liquid composition of  claim 9 , wherein the amine-containing chemical is branched polyethylenimine, cysteine, salt or derivative therefore, or any combination thereof. 
     
     
         14 . The liquid composition of  claim 11 , wherein said streptavidin serves as a linker for said gold nanoparticles (GNPs) probes and said amine-containing chemical loaded liposomes. 
     
     
         15 . The liquid composition of  claim 10 , wherein said streptavidin serves as a linker for said gold nanoparticles (GNPs) probes and said amine-containing chemical loaded liposomes, and wherein said gold nanoparticles (GNPs) probes bind to said target analyte through an antibody. 
     
     
         16 . A complex, wherein the complex comprises a target analyte moiety, a streptavidin moiety, a gold nanoparticles (GNPs) probe moiety comprising biotinylated and antibody modified gold nanoparticle (GNPs), and an amine-containing chemical loaded liposome moiety, wherein the target analyte moiety is connected to the gold nanoparticles (GNPs) probe moiety through a first antibody moiety on the gold nanoparticles, the gold nanoparticles (GNPs) probe moiety is further connected to the amine-containing chemical loaded liposome moiety through the streptavidin moiety, wherein the target analyte moiety is further connected with a second antibody that is pre-anchored on a lateral flow strip to ensure that the whole complex is anchored on the lateral flow strip.