Reagents and methods for detecting infectious diseases

Abstract

Provided herein are surface enhanced Raman scattering (SERS)-active reagents and methods for detecting one or more analyte in a sample. Said SERS-active reagents are adaptable, sensitive, and easy-to-use in the diagnosis of infectious diseases in a patient, or the detection of toxins, bacteria, viruses, pathogens, hormones, cytokines, antigens, antibodies or illicit drugs in a biological sample. Such methods may be handled by police, soldiers, or health care workers in the field, and do not require specialized training.

What is claimed: 
     
         1 . A surface enhanced Raman spectroscopy (SERS)-active reagent for detecting an analyte comprising:
 (a) one or more SERS-active surface;   (b) unmodified or modified with one or more aptamer; and   (c) one or more Raman-active marker.   
     
     
         2 . The reagent according to  claim 1 , wherein the SERS-active surface is selected from the group consisting of metals (including but not limited to silver, gold, copper, certain other transition metals and titanium nitride) semiconductor substrates (including but not limited to titanium oxide, zinc oxide, zinc selenide) or semimetals (including but not limited to graphene and molybdenum disulfide). 
     
     
         3 . The reagent according to  claim 1 , wherein the aptamer is functionalized. 
     
     
         4 . The reagent according to  claim 3 , wherein the aptamer is covalently or non-covalently attached to the SERS-active surface. 
     
     
         5 . The reagent according to  claim 1 , wherein the aptamer comprises a nucleotide sequence selected from the group consisting of SEQ ID NOs: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, and 84. 
     
     
         6 . The reagent according to  claim 1 , wherein the Raman-active marker comprises a dye, fluorescent marker, carbon nanotubes, fullerenes, alkene, alkyne, or azide. 
     
     
         7 . The reagent according to  claim 6 , wherein the Raman-active marker is a fluorescent or non-fluorescent Raman-sensitive marker, and said marker is selected from the group consisting of azides, alkynes, fluorescein (FAM), Carboxytetramethylrhodamine (TAMRA), Cy3, Texas-Red (TR), Cy3.5, Rhodamine 6G, Cy5, TRIT (tetramethyl rhodamine isothiol), NBD (7-nitrobenz-2-oxa-1,3-diazole), phthalic acid, terephthalic acid, isophthalic acid, cresyl fast violet, cresyl blue violet, brilliant cresyl blue, para-aminobenzoic acid, erythrosine, biotin, digoxigenin, 5-carboxy-4′,5′-dichloro-2′,7′-dimethoxy fluorescein, 5-carboxy-2′,4′,5′,7′-tetrachlorofluorescein, 5-carboxyfluorescein, 5-carboxy rhodamine, 6-carboxyrhodamine, 6-carboxytetramethyl amino phthalocyanines, azomethines, cyanines, xanthines, succinylfluoresceins, aminoacridine, and quantum dots. 
     
     
         8 . The reagent according to  claim 1 , wherein the aptamer undergoes a conformational change upon binding of the analyte. 
     
     
         9 . The reagent according to  claim 8 , wherein the conformational change brings the Raman-active marker into close proximity to the surface of the SERS-active surface and leads to an enhancement in the Raman signal. 
     
     
         10 . The reagent according to  claim 1 , wherein the Raman-active marker is covalently attached to the aptamer. 
     
     
         11 . The reagent according to  claim 1 , wherein the analyte is selected from the group consisting of amino acid, peptide, polypeptide, protein, glycoprotein, lipoprotein, nucleoside, nucleotide, oligonucleotide, nucleic acid, sugar, carbohydrate, oligosaccharide, polysaccharide, fatty acid, lipid, hormone, metabolite, cytokine, chemokine, receptor, neurotransmitter, antigen, allergen, antibody, substrate, metabolite, cofactor, inhibitor, drug, pharmaceutical, nutrient, prion, toxin, poison, explosive, pesticide, chemical warfare agent, biohazardous agent, radioisotope, vitamin, heterocyclic aromatic compound, carcinogen, mutagen, narcotic, amphetamine, barbiturate, hallucinogen, waste product, and contaminant. 
     
     
         12 . A diagnostic kit comprising:
 a) the SERS-active reagent of  claim 1 ;   b) at least one Raman-sensitive marker control   c) at least one positive biological fluid or tissue control; and   c) at least one negative biological fluid or tissue control.   
     
     
         13 . A detection system comprising:
 a) the SERS-active reagent of  claim 1 ; and   b) a Raman detector.   
     
     
         14 . The system according to  claim 13 , wherein the Raman detector is portable or not portable. 
     
     
         15 . The system according to  claim 13 , further comprising a sample collection apparatus. 
     
     
         16 . A method for determining the presence of one or more analyte in a biological sample, the method comprising:
 a) receiving a biological sample;   b) contacting the biological sample to at least one SERS-active reagent comprising:
 (i) one or more SERS-active surface; 
 (ii) unmodified or modified with one or more aptamer; and 
 (iii) one or more Raman-active marker; 
   c) allowing the analyte to come into contact with the aptamer;   d) binding of the analyte to the aptamer, wherein said binding causes the aptamer to undergo a conformational change;   e) irradiating the at least one SERS-active reagent bound to the one or more analyte;   f) detecting the Raman signal to generate (a) Raman spectra (um); and   g) comparing the Raman signal detected in (f) with a reference Raman signal of a control, wherein the presence of one or more analyte in the biological sample is determined when said Raman signal detected in (f) differs from said reference Raman signal.   
     
     
         17 . The method of  claim 16 , wherein an increase in the Raman signal in the Raman spectra in (f) compared to control can be correlated with the amount of the one or more analyte. 
     
     
         18 . The method of  claim 17 , wherein the conformational change in the aptamer upon binding of the analyte brings the Raman-active marker into close proximity of the SERS-active surface. 
     
     
         19 . A method for diagnosing a disease or disorder in a subject comprising the steps of:
 a) receiving a biological sample from a subject;   b) contacting the biological sample to at least one SERS-active reagent comprising:
 (i) one or more SERS-active surface; 
 (ii) unmodified or modified with one or more aptamer; and 
 (iii) one or more Raman-active marker; 
   c) allowing binding of the at least one SERS reagent to one or more analyte in the biological sample, wherein said binding causes a conformational change to the one or more aptamer of the SERS reagent;   d) irradiating the at least one SERS-active reagent bound to the one or more analyte;   e) detecting the Raman signal of the at least one SERS-active reagent; and   f) comparing the Raman signal of said at least one SERS-active reagent detected in (e) with a reference Raman signal of said at least one SERS-active reagent detected in a laboratory derived negative control sample, biological sample received from a control subject (healthy subject), or pooled biological samples from numerous control subjects (healthy subjects) wherein said disease is diagnosed when said Raman signal detected in (e) differs from said reference Raman signal in position and/or intensity.   
     
     
         20 . The method of  claim 19 , wherein the disease or disorder is selected from the group consisting of an infectious disease, proliferative disease, neurodegenerative disease, cancer, psychological disease, metabolic disease, autoimmune disease, sexually transmitted disease, gastro-intestinal disease, pulmonary disease, cardiovascular disease, stress- and fatigue-related disorder, fungal disease, pathogenic disease, and obesity-related disorder.