Methods and compositions for analyte detection

Abstract

The present invention is directed to methods and apparatus for detection of one or more analytes. Analytes include agents or components of infectious agents such as pathogenic virus, as well as enzymes, proteins and biomarkers.

1 .- 71 . (canceled) 
     
     
         72 . A method for detection of one or more analytes in a fluid sample comprising:
 (a) contacting a fluid sample with a system comprising;
 (i) a sample collection implement comprising
 (1) a swab, 
 (2) a chamber containing fluid for extracting said swab, 
 (3) a plurality of pairs of first and second detection reagents, each pair capable of specifically binding a different target analyte in said fluid sample; wherein either said first or second detection reagents further comprises a capture moiety; 
 
 (ii) a test device comprising a substrate defining a lateral flow path comprising a plurality of test regions positioned within said flow path, said test regions each containing an immobilized reagent capable of specifically binding to said capture moiety, wherein each of the test regions comprises a different immobilized reagent capable of binding a different capture moiety; and at least one control region disposed in said lateral flow path, 
 wherein said control region permits a determination of proper performance of the sensor; 
   (b) detecting whether one or more analytes bind to said immobilized capturing reagent, disposed in said test device; and   (c) determining whether one or more analytes are present in said fluid sample, if a signal is detected in at least one of said plurality of test regions.   
     
     
         73 . The method of  claim 72 , wherein said sample collection implement further comprises a reservoir for preserving a portion of said sample. 
     
     
         74 . The method of  claim 72 , where said result remains stable for visualization for a time period greater than 30 minutes. 
     
     
         75 . The method of  claim 72 , wherein said system's sensitivity, based on dilution, is equivalent to 2500 copies of influenza nucleic acid and specificity is 98% or greater. 
     
     
         76 . The method of  claim 72 , wherein said immobilized reagent is selected from the group consisting of an oligonucleotide, avidin, streptavidin, pRNA, aptamer or a combination thereof. 
     
     
         77 . The method of  claim 72 , wherein said immobilized capturing agent is pRNA, 
     
     
         78 . The method of  claim 72 , wherein each reagent in said plurality of first detection reagents comprises a label. 
     
     
         79 . The method of  claim 78 , wherein said label is selected from a group consisting of a fluorophore, a chromophore, a microparticle, a metal or a combination thereof. 
     
     
         80 . The method of  claim 78 , wherein said label is europium 
     
     
         81 . The method of  claim 72 , wherein said multiple one or more analytes are one or more viruses. 
     
     
         82 . The method of  claim 81 , wherein said one or more viruses include influenza A and/or influenza B strain. 
     
     
         83 . The method of  claim 81 , wherein said one or more viruses include different virus strains and different virus subtypes. 
     
     
         84 . The method of  claim 83 , wherein said different strains or said different subtypes include, pandemic virus, a non-pandemic virus, vaccine-available virus, vaccine-unavailable vims, or a combination thereof. 
     
     
         85 . The method of  claim 72 , wherein each of said plurality of test regions comprises said immobilized reagent specific for said capture. 
     
     
         86 .- 154 . (canceled) 
     
     
         155 . The method of  claim 72 , wherein the first and second detection reagents are separate or combined together in a pellet, a powder, a pill, a bead, a pressed lyophilized powder, or are dried on a solid support. 
     
     
         156 . The method of  claim 72 , wherein the first and second detection reagents are in solid dried form.