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 . A sample collection device comprising a body comprising:
 (a) an upper sealed chamber containing one or more solutions, and one or more breakable seals;   (b) a lower chamber in fluid communication with the upper chamber containing an extraction buffer and/or reagents;   (c) a sample collection implement in fluid communication with said upper scaled chamber; and   (d) wherein said reagents comprise a plurality of specific binding agent (SBA) sandwich pairs, each pair comprising:   (i) an SBA-capture moiety conjugate comprising an SBA which specifically binds a target antigen and a capture moiety;   (ii) an SBA-label conjugate, comprising an SHA which also specifically binds said target antigen, and wherein the pair can form a sandwich with said target antigen; and   
       wherein the plurality of pairs arc directed against different antigens. 
     
     
         2 . The sample collection device of  claim 1  wherein the capture moiety is an oligonucleotide, avidin, streptavidin, pyranosyl RNA (pRNA), aptamer or a combination thereof. 
     
     
         3 . The sample collection device of  claim 2  wherein said oligonucleotide is DNA or RNA. 
     
     
         4 . The sample collection device of  claim 1  wherein the label is a fluorophore, chromophore, metal or a combination thereof. 
     
     
         5 . The device of  claim 1 , wherein said capture moiety is pRNA and said label is Europium. 
     
     
         6 . The sample collection device of  claim 1  wherein the target antigen is one or more influenza viruses. 
     
     
         7 - 23 . (canceled) 
     
     
         24 . A sample collection device comprising a body comprising:
 (a) an upper sealed chamber containing buffer, and a valve;   (b) a lower chamber in fluid communication with the upper chamber containing (c) a swab contained within said body;   (d) a mixing area downstream of said swab comprising an adsorbent or absorbent substrate, and at least one pair of reagents comprising a label and binding moiety.   
     
     
         25 . A test device comprising a body comprising:
 (a) a lateral flow membrane in the body; and   (b) upstream of the direction of lateral flow, a chamber comprising fluid, wherein the chamber is capable of controllably releasing the fluid into the lateral flow material, wherein there is a gap between said chamber and said lateral flow membrane.   
     
     
         26 - 43 . (canceled) 
     
     
         44 . A method comprising:
 (a) forming a mixture by mixing a sample being tested for the presence of at least one target antigen with a solution comprising a plurality of reagents comprising:   (i) an antibody-capture moiety conjugate comprising an antibody which specifically binds a target antigen and a capture moiety, and   (ii) a labeled antibody, wherein the antibody specifically binds the same target antigen, wherein said reagents are directed against a plurality of different target antigens and form a complex;   (b) applying the mixture to a lateral flow membrane comprising a plurality of detection regions, each of said detection region having immobilized thereto a capture partner, each capture partner capable of specifically binding a different capture moiety;   (c) flowing the mixture across the membrane, whereby the capture moiety captures a complex having a capture moiety to which it is directed; and   (d) detecting label in at least one of said detection region, whereby detection of label indicates the presence of a target antigen in said sample.   
     
     
         45 - 85 . (canceled) 
     
     
         86 . A method for determining whether a subject is infected with one or more types of influenza virus and/or subtypes and/or strains, comprising:
 (a) contacting a sample from said subject with a system capable of detecting multiple analytes, wherein said system comprises components that are capable of specifically binding a component of one or more of said influenza virus strains and/or subtypes thus forming one or more complexes; wherein complexes formed from said -sample travel through said system via a lateral flow path, wherein said one or more complexes are capable of specifically binding an immobilized capture reagent present in two or more distinct test regions along said lateral flow path, and wherein each of said two or more test regions comprises a different said immobilized capture reagent capable of capturing a different analyte complex;   (b) determining whether said one or more influenza virus strains and subtypes are present in said fluid sample by observing said two or more test regions, wherein a positive result indicates presence of a specific Influenza virus type and/or subtype and said positive result is stable for a period greater than 30 minutes without change.   
     
     
         87 - 154 . (canceled)