WO2021247335A1

WO2021247335A1 - Viral diagnostics

Info

Publication number: WO2021247335A1
Application number: PCT/US2021/034362
Authority: WO
Inventors: Daniel Roth; Russell Hanson; George Botos
Original assignee: Aptavid Inc.
Priority date: 2020-06-04
Filing date: 2021-05-26
Publication date: 2021-12-09

Abstract

A diagnostic test including a sampling mechanism and a reaction chamber containing a solution of a nanoparticles complexed with at least one ligand that binds to a virus, wherein the apparent color of the nanoparticles change in the presence of virus. A flow-through device for detecting viruses including a housing containing a reaction chamber, wherein the reaction chamber includes a resolving layer having aptamers for the virus and control antigens embedded therein, the housing including a slot for receiving a proximal end of a funnel for applying a sample, a solution of a nanoparticle complexed with a ligand that binds to a virus, and a color developer, wherein the apparent color of the nanoparticles change in the presence of virus. A method of using the diagnostic test. A kit for detecting viruses. A surface test for detecting viruses and a method of detecting viruses on surfaces.

Description

VIRAL DIAGNOSTICS

BACKGROUND OF THE INVENTION

1. TECHNICAL FIELD

[0001] The present invention relates to compositions and methods for detecting viruses in individuals and on surfaces.

2. BACKGROUND ART

[0002] There are currently several ways in which viruses can be detected either in an individual or on surfaces of objects. Each of these tests have drawbacks to use. RT-PCR assays are used in detecting active viruses in infected patients by RNA but have the drawbacks of being an expensive lab test, they require 48 hours minimum to run, they are reagent heavy, there is risky sample pre-processing, they have a high type II error (false negatives), and they are not easily scalable. The economy during a pandemic cannot be reopened fast and safely with this approach. Antibody ELISA tests for anti-viral IgG/lgM can be used at a point of care and they detect antibodies in recovering patients, but require a blood draw at a clinic, can miss newly acquired active infections, and require trained technicians to run. These tests have further drawbacks of low sensitivity and they cannot be used on surfaces. Antibody based ELISA tests also test for viral particles also require a blood draw at a clinic and specialized personnel to run and suffer from a high false positive rate due to specificity and may also suffer from a high false negative rate due to high pH, temperature, and salt sensitivity.

[0003] There are many different assays available that provide a color or visual change upon detection of an analyte. Fluorescent labeled ligand binding assays can show binding to a target in many different colors. Bioluminescent binding assays can be used in which nanoluciferase (a bioluminescent reporter) can show binding of proteins and receptors. Bicinchoninic acid (BCA) protein assay provides a purple color in the presence of protein from a cuprous ion. Coomassie dye protein assays change from reddish-brown to blue in the presence of protein. However, many of these assays can require hours to obtain results.

[0004] Aptamer assays can be used in detecting various molecules with a color change assay.

Aptamers are short, single-stranded DNA or RNA (ssDNA or ssRNA) molecules. They selectively bind to a specific target, such as proteins, peptides, carbohydrates, small molecules, toxins, or whole cells. Aptamers can assume different shapes due to their tendency to form helices and single-stranded loops. They bind targets with high selectivity and specificity and aptamer binding is determined by its tertiary structure. Target recognition and binding involve three dimensional, shape-dependent interactions as well as hydrophobic interactions, base-stacking, and intercalation.

[0005] Mondal, et al. (Frontiers in Microbiology, February 2018, Vol. 9, Article 179) describe a colorimetric biosensor for the detection of Staphylococcal enterotoxin B (SEB) developed using SEB- binding aptamer (SEB2) as recognition element and unmodified gold nanoparticles (AuNPs) as colorimetric probes. The assay is based on color change from red to purple due to conformational change of aptamer in the presence of SEB, and the phenomenon of salt-induced AuNPs aggregation which could be monitored by naked eye or UV-vis spectrometer. Results showed that the AuNPs can effectively differentiate the SEB induced conformational change of the aptamer in the presence of a given high salt concentration. The assay was highly specific to SEB as compared to other related toxins. Robustness of the assay was tested in artificially spiked milk samples and cross-checked using in house developed sandwich ELISA (IgY as capturing and SEB specific monoclonal as revealing antibody) and PCR. Mondal, et al. state that colorimetric assays could be a suitable alternative over existing methods during biological emergencies due to its simplicity, sensitive and cost effectiveness.

[0006] Ahn, et al. (Analyst, 2009, 134, 1896-1901) have shown that an RNA aptamer can be used to detect SARS-CoV. The SARS-CoV nucleocapsid (N) protein can be a diagnostic marker for accurate and sensitive detection of the virus. Using a SELEX (systematic evolution of ligand by exponential enrichment) procedure and recombinant N protein, a high-affinity RNA aptamer was selected capable of binding to N protein with a dissociation constant of 1.65 nM. Electrophoretic mobility shift assays and RNA competition experiments showed that the selected aptamer recognized selectively the C-terminal region of N protein with high specificity. Using a chemiluminescence immunosorbent assay and a nanoarray aptamer chip with the selected aptamer as an antigen-capturing agent, N protein could be sensitively detected at a concentration as low as 2 pg/ml.

[0007] Cho, et al. (Journal of Bioscience and Bioengineering, Vol. 112, No. 6, 535-540, 2011) describe a system for detecting SARS coronavirus nucleocapsid protein using an ssDNA aptamer. An ssDNA aptamer that specifically binds to SARS CoV nucleocapsid protein was isolated from a DNA library containing 45-nuceotide random sequences in the middle of an 88mer single-stranded DNA. After twelve cycles of systematic evolution of ligands by exponential enrichment (SELEX) procedure, 15 ssDNA aptamers were identified. Enzyme-linked immunosorbent assay (ELISA) analysis was then used to identify the aptamer with the highest binding affinity to the SARS CoV nucleocapsid protein. Using this approach, an ssDNA aptamer that binds to the nucleocapsid protein with a Kd of 4.93 ± 0.30 nM was identified. Western blot analysis further demonstrated that this ssDNA aptamer could be used to efficiently detect the SARS CoV nucleocapsid protein when compared with a nucleocapsid antibody. [0008] Rapid testing devices have been developed to analyze samples for individuals thought to have infectious diseases. For example, CA2578313 (Biolytical Laboratories, Inc.) discloses diagnostic test kits including multiple recombinant antigens, of different epitopes of the same disease marker, that are capable of accurate, simultaneous, parallel detection of the presence of disease specific antibodies in a single collection of specimen applied to a single test device, thus avoiding the need to perform multiple rapid tests or to repeatedly collect specimens. The test kits can include flow-through tests or lateral- flow strips.

[0009] There remains a need for a quick and accurate test for viruses both in individuals and on surfaces that may be contaminated making use of aptamers. SUMMARY OF THE INVENTION

[00010] The present invention provides for a diagnostic test including a sampling mechanism and a reaction chamber containing a solution of a nanoparticles complexed with any number of ligands (such as aptamers, affimers, and peptides) that binds to a virus, wherein the apparent color of the nanoparticles change in the presence of virus.

[00011] The present invention provides for a flow-through device for detecting viruses including a housing containing a reaction chamber, wherein the reaction chamber includes a resolving layer having aptamers for the virus and control antigens embedded therein, the housing including a slot for receiving a proximal end of a funnel for applying a sample, a solution of a nanoparticle complexed with a ligand that binds to a virus, and a color developer, wherein the apparent color of the nanoparticles change in the presence of virus.

[00012] The present invention provides for a method of using the diagnostic test, by collecting a sample from an individual, adding the sample to the reaction chamber that captures and concentrates the virus, reacting the concentrated sample with a nanoparticle complexed with a ligand that binds to a virus, and detecting a color change in the reaction chamber, indicating the presence of virus.

[00013] The present invention also provides for a method of using the flow-through device to detect a virus, by collecting a sample from an individual with the sample collection vial having the funnel, adding the sample to the reaction chamber by placing the proximal end of the funnel in the slot of the housing wherein the reaction chamber includes aptamers for the virus and control antigens, removing the sample collection vial from the funnel, applying a vial containing a nanoparticle complexed with a ligand that binds to a virus reagent solution to the reaction chamber, applying a vial containing a color developer to the reaction chamber, and detecting a color change in the reaction chamber, indicating the presence of virus.

[00014] The present invention provides for a kit for detecting viruses, including sample collection mechanisms, a reaction chamber including a solution of a ligand complexed with a nanoparticle that binds to a virus, wherein the nanoparticle's apparent color changes in the presence of virus, and instructions for use.

[00015] The present invention also provides for a surface test for detecting viruses, including a spray mechanism including a solution of a ligand complexed with a nanoparticle that binds to a virus and changes color in the presence of virus.

[00016] The present invention provides for a surface test for detecting viruses including a swab for swiping across a surface, wherein the swab is inserted into a reaction chamber containing a solution of at least one aptamer that binds to a virus and changes color in the presence of virus.

[00017] The present invention also provides for a method of detecting viruses on surfaces, by spraying a solution of a nanoparticle complexed with a ligand that binds to a virus on a surface, wherein the nanoparticle's apparent color changes in the presence of virus on a surface and detecting a color change on the surface indicating the presence of virus.

DESCRIPTION OF THE DRAWINGS

[00018] Other advantages of the present invention are readily appreciated as the same becomes better understood by reference to the following detailed description when considered in connection with the accompanying drawings wherein:

[00019] FIGURE 1 is a schematic of the creation of aptamers with gold nanoparticles;

[00020] FIGURE 2 is a table of sequences of aptamers;

[00021] FIGURE 3 is a top perspective view of a flow-through device;

[00022] FIGURE 4 is a side perspective view of a flow-through device with a funnel and vial;

[00023] FIGURE 5 is a top perspective view of a kit;

[00024] FIGURE 6 is a photograph of color change results;

[00025] FIGURE 7 is a photograph of lateral flow test results compared to a control; [00026] FIGURE 8 is a flow chart of use of the kit with an application;

[00027] FIGURE 9A is a photograph of an application providing a timeline to the user, FIGURE 9B is a photograph of an application providing pre-test questions, FIGURE 9C is a photograph of an application providing instructions for use of the kit, and FIGURE 9D is a photograph of an application allowing for taking pictures of the test results;

[00028] FIGURE 10A is a photograph of a top view of a kit for a surface test and FIGURE 10B is a photograph of a back view of the kit;

[00029] FIGURE 11A is a diagram of a flow-through device, and FIGURE 11B is a photograph of a flow-through device;

[00030] FIGURE 12A is a photograph of a negative control, FIGURE 12B is a photograph of 1E2 vps/mL, FIGURE 12C is a photograph of 1E3 vps/mL, and FIGURE 12D is a graph showing virus particles per mL and pixel value; and

[00031] FIGURE 13A is a side perspective view of components in the reaction chamber, FIGURE 13B is a bottom view of components in the reaction chamber, FIGURE 13C is an exploded top view of components in the reaction chamber, and FIGURE 13D is an exploded bottom view of components in the reaction.

DETAILED DESCRIPTION OF THE INVENTION

[00032] The present invention provides for tests and methods for detecting the presence of viruses. More specifically, a diagnostic test includes a sampling mechanism, and a reaction chamber containing a solution of a nanoparticle and ligand complex that binds to a virus and the nanoparticle's apparent color changes in the presence of virus. The diagnostic test can provide results within 5 minutes. Most preferably, the diagnostic test is a flow-through device, further described below. A spray test for surfaces is also included and further described below.

[00033] "Virus" as used herein, refers to any infectious agent that contains a protein coat surrounding an RNA or DNA core of genetic material but no semipermeable membrane and is capable of growth and multiplication only in living cells. Most preferably, the virus in the present invention is SARS Coronavirus 2 (SARS-CoV-2), which causes COVID-19. The virus can also be, but is not limited to, adeno-associated virus, aichi virus, Australian bat lyssavirus, BK polyomavirus, banna virus, barmah forest virus, bunyamwera virus, bunyavirus La Crosse, bunyavirus snowshoe hare, cercopithecine herpesvirus, chandipura virus, chikungunya virus, cosavirus A, cowpox virus, coxsackievirus, Crimean- Congo hemorrhagic fever virus, dengue virus, dhori virus, dugbe virus, duvenhage virus, eastern equine encephalitis virus, ebolavirus, echovirus, encephalomyocarditis virus, Epstein-Barr virus, European bat lyssavirus, GB virus C/Hepatitis G virus, hantaan virus, hendra virus, hepatitis A virus, hepatitis B virus, hepatitis C virus, hepatitis E virus, hepatitis delta virus, horsepox virus, human adenovirus, human astrovirus, human coronavirus, human cytomegalovirus, human enterovirus 68, 70, human herpesvirus 1, human herpesvirus 2, human herpesvirus 6, human herpesvirus 7, human herpesvirus 8, human immunodeficiency virus, human papillomavirus 1, human papillomavirus 2, human papillomavirus 16,18, human parainfluenza, human parvovirus B19, human respiratory syncytial virus, human rhinovirus, human SARS coronavirus, human spumaretrovirus, human T-lymphotropic virus, human torovirus, influenza A virus, influenza B virus, influenza C virus, Isfahan virus, JC polyomavirus, Japanese encephalitis virus, Junin arenavirus, Kl polyomavirus, Kunjin virus, Lagos bat virus, Lake Victoria marburgvirus, langat virus, Lassa virus, lordsdale virus, louping ill virus, lymphocytic choriomeningitis virus, machupo virus, mayaro virus, MERS coronavirus, measles virus, mengo encephalomyocarditis virus, merkel cell polyomavirus, mokola virus, molluscum contagiosum virus, monkeypox virus, mumps virus, Murray valley encephalitis virus, New York virus, Nipah virus, Norwalk virus, O'nyong-nyong virus, Orf virus, Oropouche virus, Pichinde virus, poliovirus, punta toro phlebovirus, Puumala virus, rabies virus, Rift valley fever virus, rosavirus A, Ross river virus, rotavirus A, rotavirus B, rotavirus C, rubella virus,

Sagiyama virus, Salivirus A, sandfly fever Sicilian virus, Sapporo virus, Semliki forest virus, Seoul virus, simian foamy virus, simian virus 5, Sindbis virus, Southampton virus, St. Louis encephalitis virus, tick- borne powassan virus, Torque teno virus, Toscana virus, Uukuniemi virus, Vaccinia virus, varicella-zoster virus, Variola virus, Venezuelan equine encephalitis virus, vesicular stomatitis virus, western equine encephalitis virus, WU polyomavirus, West Nile virus, Yaba monkey tumor virus, Yaba-like disease virus, yellow fever virus, or Zika virus.

[00034] "Sample" as used herein, can refer to any collection of particles/fluids from an individual, including saliva, tears, nasal cavity, blood, plasma, urine, breath, or a sample from a swab to any part of the body. Samples can also be taken from any surface of an object.

[00035] The virus particles for SARS-CoV-2 consist of primarily three components: the spike protein, the nucleocapsid, and the coding mRNA. Wrapp, et al. (Science, 367, 1260-1263 (2020)) have shown the cryo-EM structure of the spike protein as well as binding conformations with host cells. Each chemical assay is designed to test for one or more of these components, or several of the viral pieces simultaneously. Testing for the nucleocapsid is not commonly emphasized. In some cases the binding domain of a ligand on the spike protein may be blocked by the nucleocapsid, though smaller ligands will have fewer issues with blocked sites.

[00036] The ligands complexed with the nanoparticle can be, but are not limited to, aptamers, affimers, or peptides. A single ligand can be used or multiple ligands can be used in the complex. Preferably, the diagnostic test uses aptamers bound to a nanoparticle in a colloidal solution. An aptamer is a molecule that is engineered to bind to other molecules in very specific ways. An aptamer can be created to bind to the SARS-CoV-2 virus, or any other virus desired to be detected. By attaching a gold nanoparticle to the aptamer, when virus particles bind to the aptamer, the color of the solution changes. It should be understood that the nanoparticles do not actually change color, rather, it is more of an optical illusion provided by the aggregation of particles that provides the color change. This process is shown in FIGURE 1. The gold nanoparticles are preferably between 5 and 15 nm in size, however smaller (1-5 nm) or larger (15-50 nm) nanoparticles can also be used. Alternatively, affimers are small proteins that bind to target molecules with similar specificity and affinity to that of antibodies. They are engineered non-antibody binding proteins that are designed to mimic the molecular recognition characteristics of monoclonal antibodies. Peptides are short chains of two to fifty amino acids linked by peptide bonds and can be used to bind to other molecules.

[00037] The shape of nanoparticle is preferably star-shaped, however other shapes (round, rod, hollow, etc.) can be used. The advantage of star-shaped nanoparticles is that the color of the non- aggregated suspension is blue and will shift to red through aggregation/ attachment to larger particles such as virus.

[00038] In determining aptamers for SARS-CoV-2, the first eight rounds of screening followed classic protein SELEX protocols to ensure that the DNA strands with SARS-CoV-2 RBD binding capacity amplify to multiple copies. After 8 rounds of standard selection procedures, ACE2 competition was included in the next 4 rounds of selection. After the DNA library was incubated with RBD-beads, the unbound or weakly bound sequences in the supernatant were removed. The K_d values of the optimized CoV2-RBD-lC and CoV2-RBD-4C aptamers against RBD were 5.8 nM and 19.9 nM, respectively.

[00039] FIGURE 2 shows sequences of selected aptamers for SARS-CoV-2 with the bold sequences being primers (SEQ ID NOs: 1-8).

[00040] The advantage of using a colloidal solution is that the reaction tends to happen faster due to higher mobility of both the virus and the nanoparticles. The reaction chamber allows one to directly suspend, in one step, the sample into the reagent solution, without the need for additional wash steps, with a minimal amount of reagent (0.1 to 0.5 ml). The design of the reaction chamber can be between a micro-cuvette on the lower side and a reaction chamber with a swab in the cap for sample collection. The reaction can also be performed inside a pipet tip and a bulb can be used to mix. Another design for the reaction chamber is a plastic ampoule in a vial design, with a cap with the swab built in. [00041] Most preferably, the diagnostic test 10 is a flow-through device 12 including a housing 14 containing a reaction chamber 16, shown in FIGURE 3, to which various vials 22 can be applied to provide samples and additional reagents and chemicals. In general, flow-through assays allow for detection of a chemical compound from a sample when it interacts with another chemical compound bound to a reactive membrane panel, while the rest of the sample flows through to an absorbent pad. A signal reagent can be applied to the reactive membrane panel to provide a visual presence of the bound chemical compound present in the sample. A control can also be provided on the membrane, wherein a colored line or dot can indicate that the test has been performed correctly.

[00042] The housing 14 is preferably made of plastic but can be other materials. The reaction chamber 16 contains a resolving layer 18 (preferably of nitrocellulose membrane) that can have either aptamers (Test) or antigens (Control) applied thereto that react with the sample.

[00043] A sample collection vial 22 is provided along with a funnel 24 to aid the user in depositing a saliva sample into the sample collection vial 22, shown in FIGURE 4. The vial 22 and the funnel 24 can be made of plastic or any suitable material. A distal end 26 of the funnel 24 is threaded for removable attachment to various vials 22 (i.e. the sample collection vial 22 includes a proximal end 30 that is also threaded), and also includes a puncture mechanism 36. A proximal end 28 of the funnel is flared and fits in a slot 32 on the housing 14 over the reaction chamber 16. The sample collection vial 22 or any other vial 22 can include a cap 34 for sealing when not in use. Use of a flow-through device 12 can be beneficial when needing to analyze a large sample volume (1 ml saliva). The sample of saliva containing virus can have detergents added to improve flow and solubilize the virus such as tween and triton, NP40, or deoxycholate, and any such compounds can be included in additional vials 22 for addition to the sample collection vial 22. Once the saliva sample is collected, the sample collection vial 22 is applied to the reaction chamber 16 by tipping the sample collection vial 22 upside down and placing the proximal end

28 of the funnel 24 in the slot 32. The saliva then flows down into the reaction chamber 16. The sample collection vial 22 can then be removed from the funnel 24 by unscrewing the proximal end 30 from the distal end 26 of the funnel 24.

[00044] After applying the sample, additional reagents and chemicals can be applied to the reaction chamber 16. For example, a vial 22 containing a nanoparticle reagent solution can next be applied to the reaction chamber 16. Preferably, the vial 22 containing the reagent solution includes a seal 38 at the proximal end 30 to preserve the chemicals until use that can be made of foil or any other suitable material. When ready to use, the proximal end 30 is engaged with the puncture mechanism 36 by screwing the vial 22 onto the funnel 24. This opens the seal 38 and the reagent is released from the vial 22 into the reaction chamber 16. Next a vial 22 containing a color developer can be applied to the reaction chamber 16 in the same manner, by engaging proximal end 30 of the vial 22 with the puncture mechanism 36 of the funnel 24 to release the color developer into the reaction chamber 16.

[00045] After all necessary reagents or chemicals have been added to the reaction chamber 16, the reaction with the sample can then proceed. The funnel 24 can be removed from the slot 32 and the results of the reaction can be read within the reaction chamber 16 wherein a color change indicates the presence of a virus. FIGURE 6 shows an example of a color change in the presence of various amounts of SARS-CoV-2, wherein a negative sample does not change color and a positive sample changes to a pink color. FIGURE 7 is an example of lateral flow test results compared to a control.

[00046] A more detailed view of the components of the reaction chamber 16 is shown in FIGURES 13A-13D. A membrane holding portion 50 includes a recess 52 for holding the resolving layer 18 and a wick 54 underneath. In other words, the resolving layer 18 rests on top of the wick 54, and a portion of the resolving layer 18 is exposed to receive samples and chemicals in the reaction chamber 16. The wick 54 can extend past the membrane holding portion 50 in either direction and can be covered by the housing 14. A top portion 56 receiving funnel 24 fits on top of the membrane holding portion 50 and the two portions 56, 50 are secured together with clips 60 which can slide into place around the two portions 56, 50. Funnel 24 can snap into the top portion 56 with a friction fit and tabs. Example materials of the portions 50, 56 and clips 60 include plastics such as polylactic acid (PLA) or polyethylene terephthalate glycol (PETG).

[00047] The flow-through device 12 provides test results in 5 minutes with a 10-100 TCID50 /mL limit of detection. The flow-through device 12 can be sealed in protective packaging 20 until use. [00048] Preferably, the nanoparticles functionalized with the ligand attached to the virus surface leads to nanoparticle aggregation or deaggregation and color change. However, if the color change is not intense enough to be easily observed (or for increasing the sensitivity of the test) a secondary nanoparticle reagent can be used that are larger that the viral particles and are designed to have a functionalized surface (positively or negatively charged) that can aggregate a large number of the non- reacted nanoparticles per particle and produce a stronger color change. Secondary oligonucleotides can also cause or prevent aggregation.

[00049] The flow-through device 12 can also be used in conjunction with an application ("app") for tablets and smartphones stored on non-transitory computer readable media. The protective packaging 20 as well as the housing 14 can include a code 42 (QR code or barcode) unique to the particular device 12. FIGURE 8 shows a flowchart of use of the flow-through device 12 with the application. The user registers the device 12/kit 40 with the code 42 by scanning the code 42 with the application, samples are collected, the results in the reaction chamber can be captured with the application and sent to healthcare professionals for review and consultation, and then the application can provide automated follow ups and reminders to the user. The application can provide a timeline to the user (FIGURE 9A), provide pre-test questions (FIGURE 9B), provide instructions for use of the kit (FIGURE 9C), and allow for taking pictures of the test results (FIGURE 9D).

[00050] The present invention provides for a method of using the diagnostic test, by collecting a sample from an individual, adding the sample to the reaction chamber, reacting the sample with a nanoparticle complexed with a ligand that binds to a virus, and detecting a color change in the reaction chamber, indicating the presence of virus. Preferably, the color changes from blue to red in the presence of virus as described above. However, other colors can also be used.

[00051] The present invention also provides for a method of using the flow-through device 10 to detect a virus, by collecting a sample from an individual with the sample collection vial 22 having the funnel 24, adding the sample to the reaction chamber 16 by placing the proximal end 28 of the funnel 24 in the slot 32 of the housing 14 wherein the reaction chamber 16 includes aptamers for the virus and control antigens, removing the sample collection vial 22 from the funnel 24, applying a vial 22 containing the nanoparticle complexed with a ligand that binds to a virus reagent solution to the reaction chamber 16, applying a vial 22 containing a color developer to the reaction chamber 16, and detecting a color change in the reaction chamber 16, indicating the presence of virus. As described above, the vials 22 can be screwed onto the distal end 26 of the funnel 24 and the puncture mechanism 36 can break any seal on the vials 22 allowing the contents to flow to the reaction chamber 16.

[00052] The present invention provides for a kit 40 for detecting viruses, including sample collection mechanisms (such as swabs, test tubes, syringes), a reaction chamber including the solution of a nanoparticle complexed with a ligand that binds to a virus and wherein the nanoparticle's apparent color changes in the presence of virus, and instructions for use. More specifically, as shown in FIGURE 5, the kit 40 can include a flow-through device 12, vials 22 for collecting a saliva sample as well as a nanoparticle reagent solution and a color developer, and the funnel 24. Personal protective equipment such as gloves and face masks can be optionally included. Any necessary packaging can also be included. [00053] The present invention also provides for a surface test 100 for detecting viruses, including a spray mechanism including a solution of a nanoparticle complexed with a ligand that binds to a virus and wherein the nanoparticle's apparent color changes in the presence of virus. The spray mechanism can be a spray bottle including the solution. When the solution is sprayed onto a surface that has a virus on it, the solution changes color. The color change can be detected by eye (such as the change from blue to red as above) or by shining a fluorescent or ultraviolet light on the surface. Qdots and quenchers (such as riboflavin quenchers) can also be used. Alternatively, the surface test 100 can use a swab 102 as in the diagnostic test that is swiped across a surface and then (at least the part that has contacted the surface) inserted into a reaction chamber 104 containing a solution of at least one aptamer that binds to a virus and changes color in the presence of virus (the reagents and other chemicals can be contained in a separate vial 106 to be added to the reaction chamber 104). The surface test is especially useful in detecting viruses on tables, desks, doors, door handles, mobile device screens, menus, sinks, bathrooms, or any other surface commonly touched. This can help companies determine what surfaces need to be cleaned better. A kit 108 can be provided including the swab 102, reaction chamber 104, additional vials 106 for reagents, and instructions for use, as shown in FIGURES 10A and 10B.

[00054] The present invention also provides for a method of detecting viruses on surfaces, by spraying a solution of a nanoparticle complexed with a ligand that binds to a virus and wherein the nanoparticle's apparent color changes in the presence of virus on a surface and detecting a color change on the surface indicating the presence of virus. The color changes from blue to red in the presence of virus as described above (which would be easiest to detect on a white or light colored surface). However, other colors can also be used in order to detect the colors on other colored surfaces. Alternatively, a swab is swiped across a surface and inserted into a reaction chamber containing a solution of the nanoparticle complexed to the ligand that binds to a virus and changes color in the presence of virus, and a color change is detected in the presence of virus.

[00055] The diagnostic test and spray test of the present invention provide advantages over previous tests for viruses because the test detects the virus particles themselves and can be used for on site, at-home, or at-worksite testing. The diagnostic test of the present invention does not require incubation at 65 degrees C or thermocycling as in RT-PCR assays, or an inherent delay as the immune system of the individual develops a response to the viral particles themselves while fighting the disease. [00056] The invention is further described in detail by reference to the following experimental examples. These examples are provided for the purpose of illustration only and are not intended to be limiting unless otherwise specified. Thus, the invention should in no way be construed as being limited to the following examples, but rather, should be construed to encompass any and all variations which become evident as a result of the teaching provided herein.

[00057] EXAMPLE 1

[00058] The following was performed in preparing the flow-through device 12 and testing.

[00059] TABLE 1: The preparation of reagents/mixture used in the flow through experiment.

[00060] SARS-CoV-2 viral lysate is firstly thawed and then prepared by diluting the stock solution to desired concentrations in CL2 lab.

[00061] A) Immobilization of capture aptamer on NC membrane:

[00062] 1) A mix was prepared containing streptavidin and biotinylated capture aptamer (Nuc 1) in

1:9 molar ratio, this is to make sure all biotin receptor sites are occupied.

[00063] 2) This was incubated at 37 degree Celsius for 30 minutes in water bath or heating block.

[00064] 3) 1 uL of the above solution was spotted onto the nitrocellulose membrane.

[00065] 4) The spot was allowed to dry on the membrane, at 37 degree Celsius for 1 hour in an incubator.

[00066] B) Conjugation of biotinylated detection aptamer to HRP-streptavidin: [00067] 1) 200 pmoles of biotinylated detection aptamer (Nuc 2) was conjugated with a 1:2.5 dilution of HRP-streptavidin in a total volume of 50 uL (in buffer) for at least 30 minutes (TABLE 1). Detecting enzyme HRP binds to the detection aptamer by non-covalent interactions between streptavidin and biotin.

[00068] 2) Target protein (viral lysate) was added at desired concentration to the running buffer.

[00069] 3) 50 uL of the aptamer/HRP mixture from step 1 was added to the running buffer from step 2. The total volume of running buffer + target protein + detection aptamer/HRP should be 1 mL. The solution is incubated for about 15 minutes at RT.

[00070] Part C) Testing of presence of target protein by HRP-TMB color development method:

[00071] 1) The dried membrane is taken out from incubator and the flow through apparatus is assembled as shown in FIGURES 11A and 11B.

[00072] 2) 1 mL of running buffer containing detection aptamer bound to HRP and target protein is loaded into the funnel. The solution is slowly passed through the flow through device, this process may take up to 5 minutes.

[00073] 3) The membrane is taken out and put it aside, the wicking pads are changed in the apparatus and it is reassembled by placing the membrane back onto a newly replaced dry square wicking pad.

[00074] 4) 1 mL of chase buffer is added to the funnel, waiting for it to pass through completely.

The purpose of this washing step is to wash off any free/unbound HRP-streptavidin from the membrane. [00075] 5) Once all the chase buffer passes through the apparatus, the membrane is taken out and placed in 500 uL of diluted TMB substrate solution (dilution of TMB is 1:3). The membrane stays in the solution and incubates for about 1 - 2 minutes until color develops.

[00076] Positive results: A bright blue colour in the test zone indicates the presence of target protein. [00077] EXAMPLE 2

[00078] A pixel analysis was performed with the presence and absence of lysate. FIGURE 12A shows a negative control, FIGURE 12B shows 1E2 vps/mL, and FIGURE 12C shows 1E3 vps/mL. FIGURE 12D is a graph showing virus particles per mL and pixel value. The amount of contrast, as measured by the pixel value is a measure of how much nanoparticle and bound virus/virus antigen is present in the patient saliva sample. The nanoparticles are a reporter of virus presence. The darker the spot on the nitrocellulose the more virus antigen is present in the sample.

[00079] Throughout this application, various publications, including United States patents, are referenced by author and year and patents by number. Full citations for the publications are listed below. The disclosures of these publications and patents in their entireties are hereby incorporated by reference into this application in order to more fully describe the state of the art to which this invention pertains.

[00080] The invention has been described in an illustrative manner, and it is to be understood that the terminology, which has been used is intended to be in the nature of words of description rather than of limitation.

[00081] Obviously, many modifications and variations of the present invention are possible in light of the above teachings. It is, therefore, to be understood that within the scope of the appended claims, the invention can be practiced otherwise than as specifically described.

Claims

CLAIMS What is claimed is:

1. A diagnostic test for detecting the presence of viruses, comprising: a sampling mechanism; and a reaction chamber containing a solution of a nanoparticles complexed with at least one ligand that binds to a virus, wherein an apparent color of said nanoparticles change in the presence of said virus.

2. The diagnostic test of claim 1, wherein said virus is chosen from the group consisting of SARS Coronavirus 2 (SARS-CoV-2), adeno-associated virus, aichi virus, Australian bat lyssavirus, BK polyomavirus, banna virus, barmah forest virus, bunyamwera virus, bunyavirus La Crosse, bunyavirus snowshoe hare, cercopithecine herpesvirus, chandipura virus, chikungunya virus, cosavirus A, cowpox virus, coxsackievirus, Crimean-Congo hemorrhagic fever virus, dengue virus, dhori virus, dugbe virus, duvenhage virus, eastern equine encephalitis virus, ebolavirus, echovirus, encephalomyocarditis virus, Epstein-Barr virus, European bat lyssavirus, GB virus C/Hepatitis G virus, hantaan virus, hendra virus, hepatitis A virus, hepatitis B virus, hepatitis C virus, hepatitis E virus, hepatitis delta virus, horsepox virus, human adenovirus, human astrovirus, human coronavirus, human cytomegalovirus, human enterovirus 68, 70, human herpesvirus 1, human herpesvirus 2, human herpesvirus 6, human herpesvirus 7, human herpesvirus 8, human immunodeficiency virus, human papillomavirus 1, human papillomavirus 2, human papillomavirus 16,18, human parainfluenza, human parvovirus B19, human respiratory syncytial virus, human rhinovirus, human SARS coronavirus, human spumaretrovirus, human T-lymphotropic virus, human torovirus, influenza A virus, influenza B virus, influenza C virus, Isfahan virus, JC polyomavirus, Japanese encephalitis virus, Junin arenavirus, Kl polyomavirus, Kunjin virus, Lagos bat virus, Lake Victoria marburgvirus, langat virus, Lassa virus, lordsdale virus, louping ill virus, lymphocytic choriomeningitis virus, machupo virus, mayaro virus, MERS coronavirus, measles virus, mengo encephalomyocarditis virus, merkel cell polyomavirus, mokola virus, molluscum contagiosum virus, monkeypox virus, mumps virus, Murray valley encephalitis virus, New York virus, Nipah virus, Norwalk virus, O'nyong-nyong virus, Orf virus, Oropouche virus, Pichinde virus, poliovirus, punta toro phlebovirus, Puumala virus, rabies virus, Rift valley fever virus, rosavirus A, Ross river virus, rotavirus A, rotavirus B, rotavirus C, rubella virus, Sagiyama virus, Salivirus A, sandfly fever Sicilian virus, Sapporo virus, Semliki forest virus, Seoul virus, simian foamy virus, simian virus 5, Sindbis virus, Southampton virus, St. Louis encephalitis virus, tick- borne powassan virus, Torque teno virus, Toscana virus, Uukuniemi virus, Vaccinia virus, varicella-zoster virus, Variola virus, Venezuelan equine encephalitis virus, vesicular stomatitis virus, western equine encephalitis virus, WU polyomavirus, West Nile virus, Yaba monkey tumor virus, Yaba-like disease virus, yellow fever virus, and Zika virus.

3. The diagnostic test of claim 1, wherein said sampling mechanism is chosen from the group consisting of a swab, test tube, syringe, and vial.

4. The diagnostic test of claim 1, wherein said sampling mechanism receives a sample chosen from the group consisting of saliva, tears, nasal cavity, blood, plasma, urine, breath, and a sample from a swab to any part of the body.

5. The diagnostic test of claim 1, wherein said at least one ligand is chosen from the group consisting of aptamers, affimers, and peptides.

6. The diagnostic test of claim 1, wherein said virus is SARS-CoV-2 and said at least one ligand is an aptamer chosen from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, and combinations thereof.

7. The diagnostic test of claim 1, wherein said nanoparticles are further defined as gold nanoparticles 5 to 15 nm in size.

8. The diagnostic test of claim 1, wherein said nanoparticles are in a shape chosen from the group consisting of star, round, rod, and hollow.

9. A flow-through device for detecting viruses, comprising: a housing containing a reaction chamber, wherein said reaction chamber includes a resolving layer having aptamers for a virus and control antigens embedded therein, said housing including a slot for receiving a proximal end of a funnel for applying a sample, a solution of a nanoparticle complexed with a ligand that binds to a virus, and a color developer, and wherein an apparent color of said nanoparticles change in the presence of said virus.

10. The flow-through device of claim 9, further including a sample collection vial that is attachable to a distal end of said funnel through threading.

11. The flow-through device of claim 9, wherein said sample collection vial further includes a cap.

12. The flow-through device of claim 9, wherein said distal end of said funnel includes a puncture mechanism for opening seals on vials.

13. The flow-through device of claim 9, wherein said proximal end of said funnel is flared and fits in said slot over said reaction chamber.

14. The flow-through device of claim 9, further including vials of detergents for addition to said sample collection vial.

15. The flow-through device of claim 9, wherein said reaction chamber includes a membrane holding portion having a recess for holding said resolving layer and holding a wick underneath said resolving layer.

16. The flow-through device of claim 15, wherein said reaction chamber further includes a top portion that receives said funnel on top of said membrane holding portion and said top portion and said membrane holding portion are secured together with clips.

17. The flow-through device of claim 9, wherein said flow-through device is sealed in protective packaging.

18. The flow-through device of claim 17, wherein said protective packaging includes a code unique to said device for use with an application stored on non-transitory computer readable media.

19. The flow-through device of claim 9, wherein said virus is chosen from the group consisting of SARS Coronavirus 2 (SARS-CoV-2), adeno-associated virus, aichi virus, Australian bat lyssavirus, BK polyomavirus, banna virus, barmah forest virus, bunyamwera virus, bunyavirus La Crosse, bunyavirus snowshoe hare, cercopithecine herpesvirus, chandipura virus, chikungunya virus, cosavirus A, cowpox virus, coxsackievirus, Crimean-Congo hemorrhagic fever virus, dengue virus, dhori virus, dugbe virus, duvenhage virus, eastern equine encephalitis virus, ebolavirus, echovirus, encephalomyocarditis virus, Epstein-Barr virus, European bat lyssavirus, GB virus C/Hepatitis G virus, hantaan virus, hendra virus, hepatitis A virus, hepatitis B virus, hepatitis C virus, hepatitis E virus, hepatitis delta virus, horsepox virus, human adenovirus, human astrovirus, human coronavirus, human cytomegalovirus, human enterovirus 68, 70, human herpesvirus 1, human herpesvirus 2, human herpesvirus 6, human herpesvirus 7, human herpesvirus 8, human immunodeficiency virus, human papillomavirus 1, human papillomavirus 2, human papillomavirus 16,18, human parainfluenza, human parvovirus B19, human respiratory syncytial virus, human rhinovirus, human SARS coronavirus, human spumaretrovirus, human T-lymphotropic virus, human torovirus, influenza A virus, influenza B virus, influenza C virus, Isfahan virus, JC polyomavirus, Japanese encephalitis virus, Junin arenavirus, Kl polyomavirus, Kunjin virus, Lagos bat virus, Lake Victoria marburgvirus, langat virus, Lassa virus, lordsdale virus, louping ill virus, lymphocytic choriomeningitis virus, machupo virus, mayaro virus, MERS coronavirus, measles virus, mengo encephalomyocarditis virus, merkel cell polyomavirus, mokola virus, molluscum contagiosum virus, monkeypox virus, mumps virus, Murray valley encephalitis virus, New York virus, Nipah virus, Norwalk virus, O'nyong-nyong virus, Orf virus, Oropouche virus, Pichinde virus, poliovirus, punta toro phlebovirus, Puumala virus, rabies virus, Rift valley fever virus, rosavirus A, Ross river virus, rotavirus A, rotavirus B, rotavirus C, rubella virus, Sagiyama virus, Salivirus A, sandfly fever Sicilian virus, Sapporo virus, Semliki forest virus, Seoul virus, simian foamy virus, simian virus 5, Sindbis virus, Southampton virus, St. Louis encephalitis virus, tick- borne powassan virus, Torque teno virus, Toscana virus, Uukuniemi virus, Vaccinia virus, varicella-zoster virus, Variola virus, Venezuelan equine encephalitis virus, vesicular stomatitis virus, western equine encephalitis virus, WU polyomavirus, West Nile virus, Yaba monkey tumor virus, Yaba-like disease virus, yellow fever virus, and Zika virus.

20. The flow-through device of claim 9, wherein said sample is chosen from the group consisting of saliva, tears, nasal cavity, blood, plasma, urine, breath, and a sample from a swab to any part of the body.

21. The flow-through device of claim 9, wherein said at least one ligand is chosen from the group consisting of aptamers, affimers, and peptides.

22. The flow-through device of claim 9, wherein said virus is SARS-CoV-2 and said at least one ligand is an aptamer chosen from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, and combinations thereof.

23. The flow-through device of claim 9, wherein said nanoparticles are further defined as gold nanoparticles 5 to 15 nm in size.

24. The flow-through device of claim 9, wherein said nanoparticles are in a shape chosen from the group consisting of star, round, rod, and hollow.

25. A method of using a diagnostic test for detection of viruses, including the steps of: collecting a sample from an individual; adding the sample to a reaction chamber that captures and concentrates the virus; reacting the concentrated sample with a nanoparticle complexed with a ligand that binds to the virus; and detecting a color change in the reaction chamber, indicating the presence of virus.

26. The method of claim 25, wherein the virus is chosen from the group consisting of SARS

Coronavirus 2 (SARS-CoV-2), adeno-associated virus, aichi virus, Australian bat lyssavirus, BK polyomavirus, banna virus, barmah forest virus, bunyamwera virus, bunyavirus La Crosse, bunyavirus snowshoe hare, cercopithecine herpesvirus, chandipura virus, chikungunya virus, cosavirus A, cowpox virus, coxsackievirus, Crimean-Congo hemorrhagic fever virus, dengue virus, dhori virus, dugbe virus, duvenhage virus, eastern equine encephalitis virus, ebolavirus, echovirus, encephalomyocarditis virus, Epstein-Barr virus, European bat lyssavirus, GB virus C/Hepatitis G virus, hantaan virus, hendra virus, hepatitis A virus, hepatitis B virus, hepatitis C virus, hepatitis E virus, hepatitis delta virus, horsepox virus, human adenovirus, human astrovirus, human coronavirus, human cytomegalovirus, human enterovirus 68, 70, human herpesvirus 1, human herpesvirus 2, human herpesvirus 6, human herpesvirus 7, human herpesvirus 8, human immunodeficiency virus, human papillomavirus 1, human papillomavirus 2, human papillomavirus 16,18, human parainfluenza, human parvovirus B19, human respiratory syncytial virus, human rhinovirus, human SARS coronavirus, human spumaretrovirus, human T-lymphotropic virus, human torovirus, influenza A virus, influenza B virus, influenza C virus, Isfahan virus, JC polyomavirus, Japanese encephalitis virus, Junin arenavirus, Kl polyomavirus, Kunjin virus, Lagos bat virus, Lake Victoria marburgvirus, langat virus, Lassa virus, lordsdale virus, louping ill virus, lymphocytic choriomeningitis virus, machupo virus, mayaro virus, MERS coronavirus, measles virus, mengo encephalomyocarditis virus, merkel cell polyomavirus, mokola virus, molluscum contagiosum virus, monkeypox virus, mumps virus, Murray valley encephalitis virus, New York virus, Nipah virus, Norwalk virus, O'nyong-nyong virus, Orf virus, Oropouche virus, Pichinde virus, poliovirus, punta toro phlebovirus, Puumala virus, rabies virus, Rift valley fever virus, rosavirus A, Ross river virus, rotavirus A, rotavirus B, rotavirus C, rubella virus, Sagiyama virus, Salivirus A, sandfly fever Sicilian virus, Sapporo virus, Semliki forest virus, Seoul virus, simian foamy virus, simian virus 5, Sindbis virus, Southampton virus, St. Louis encephalitis virus, tick- borne powassan virus, Torque teno virus, Toscana virus, Uukuniemi virus, Vaccinia virus, varicella-zoster virus, Variola virus, Venezuelan equine encephalitis virus, vesicular stomatitis virus, western equine encephalitis virus, WU polyomavirus, West Nile virus, Yaba monkey tumor virus, Yaba-like disease virus, yellow fever virus, and Zika virus.

27. The method of claim 25, wherein the sample is chosen from the group consisting of saliva, tears, nasal cavity, blood, plasma, urine, breath, and a sample from a swab to any part of the body.

28. The method of claim 25, wherein the at least one ligand is chosen from the group consisting of aptamers, affimers, and peptides.

29. The method of claim 25, wherein the virus is SARS-CoV-2 and the at least one ligand is an aptamer chosen from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, and combinations thereof.

30. The method of claim 25, wherein the nanoparticles are further defined as gold nanoparticles 5 to 15 nm in size.

31. The method of claim 25, wherein the nanoparticles are in a shape chosen from the group consisting of star, round, rod, and hollow.

32. A method of using a flow-through device to detect a virus, including the steps of: collecting a sample from an individual with a sample collection vial including a funnel; adding the sample to a reaction chamber in the flow-through device by placing a proximal end of the funnel in a slot of a housing of the flow-through device wherein the reaction chamber includes aptamers for the virus and control antigens; removing the sample collection vial from the funnel; applying a vial containing a nanoparticle complexed with a ligand that binds to a virus reagent solution to the reaction chamber; applying a vial containing a color developer to the reaction chamber; and detecting a color change in the reaction chamber, indicating the presence of virus.

33. The method of claim 32, wherein said removing the sample collection vial step is further defined as unscrewing a proximal end of the sample collection vial from a distal end of the funnel.

34. The method of claim 32, wherein said collecting a sample step further includes the step of adding detergents to improve flow and solubilize the virus.

35. The method of claim 32, wherein said applying a vial containing a virus reagent solution step is further defined as engaging a proximal end of the vial with a puncture mechanism of the funnel by screwing the vial onto the funnel and releasing the virus reagent solution into the reaction chamber.

36. The method of claim 32, wherein said applying a vial containing a color developer step is further defined as engaging a proximal end of the vial with a puncture mechanism of the funnel by screwing the vial onto the funnel and releasing the color developer into the reaction chamber.

37. The method of claim 32, further including, after said applying a vial containing a color developer step, the step of removing the funnel from the slot.

38. The method of claim 32, further including, before said collecting step, the step of registering the flow-through device with an application on non-transitory computer readable media by scanning a code on protective packaging.

39. The method of claim 38, further including the step of capturing results with the application in the reaction chamber and sending the results to healthcare professionals.

40. The method of claim 39, further including the steps of the application providing a timeline to the individual, providing pre-test questions, and providing instructions for use of the flow-through device.

41. The method of claim 32, wherein the virus is chosen from the group consisting of SARS Coronavirus 2 (SARS-CoV-2), adeno-associated virus, aichi virus, Australian bat lyssavirus, BK polyomavirus, banna virus, barmah forest virus, bunyamwera virus, bunyavirus La Crosse, bunyavirus snowshoe hare, cercopithecine herpesvirus, chandipura virus, chikungunya virus, cosavirus A, cowpox virus, coxsackievirus, Crimean-Congo hemorrhagic fever virus, dengue virus, dhori virus, dugbe virus, duvenhage virus, eastern equine encephalitis virus, ebolavirus, echovirus, encephalomyocarditis virus,

Epstein-Barr virus, European bat lyssavirus, GB virus C/Hepatitis G virus, hantaan virus, hendra virus, hepatitis A virus, hepatitis B virus, hepatitis C virus, hepatitis E virus, hepatitis delta virus, horsepox virus, human adenovirus, human astrovirus, human coronavirus, human cytomegalovirus, human enterovirus 68, 70, human herpesvirus 1, human herpesvirus 2, human herpesvirus 6, human herpesvirus 7, human herpesvirus 8, human immunodeficiency virus, human papillomavirus 1, human papillomavirus 2, human papillomavirus 16,18, human parainfluenza, human parvovirus B19, human respiratory syncytial virus, human rhinovirus, human SARS coronavirus, human spumaretrovirus, human T-lymphotropic virus, human torovirus, influenza A virus, influenza B virus, influenza C virus, Isfahan virus, JC polyomavirus, Japanese encephalitis virus, Junin arenavirus, Kl polyomavirus, Kunjin virus, Lagos bat virus, Lake Victoria marburgvirus, langat virus, Lassa virus, lordsdale virus, louping ill virus, lymphocytic choriomeningitis virus, machupo virus, mayaro virus, MERS coronavirus, measles virus, mengo encephalomyocarditis virus, merkel cell polyomavirus, mokola virus, molluscum contagiosum virus, monkeypox virus, mumps virus, Murray valley encephalitis virus, New York virus, Nipah virus, Norwalk virus, O'nyong-nyong virus, Orf virus, Oropouche virus, Pichinde virus, poliovirus, punta toro phlebovirus, Puumala virus, rabies virus, Rift valley fever virus, rosavirus A, Ross river virus, rotavirus A, rotavirus B, rotavirus C, rubella virus, Sagiyama virus, Salivirus A, sandfly fever Sicilian virus, Sapporo virus, Semliki forest virus, Seoul virus, simian foamy virus, simian virus 5, Sindbis virus, Southampton virus, St. Louis encephalitis virus, tick- borne powassan virus, Torque teno virus, Toscana virus, Uukuniemi virus, Vaccinia virus, varicella-zoster virus, Variola virus, Venezuelan equine encephalitis virus, vesicular stomatitis virus, western equine encephalitis virus, WU polyomavirus, West Nile virus, Yaba monkey tumor virus, Yaba-like disease virus, yellow fever virus, and Zika virus.

42. The method of claim 32, wherein the sample is chosen from the group consisting of saliva, tears, nasal cavity, blood, plasma, urine, breath, and a sample from a swab to any part of the body.

43. The method of claim 32, wherein the ligand is chosen from the group consisting of aptamers, affimers, and peptides.

44. The method of claim 32, wherein the virus is SARS-CoV-2 and the at least one ligand is an aptamer chosen from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, and combinations thereof.

45. The method of claim 32, wherein the nanoparticles are further defined as gold nanoparticles 5 to 15 nm in size.

46. The method of claim 32, wherein the nanoparticles are in a shape chosen from the group consisting of star, round, rod, and hollow.

47. A kit for detecting viruses, comprising: sample collection mechanisms; a reaction chamber including a solution of a ligand complexed with a nanoparticle that binds to a virus, wherein the nanoparticle's apparent color changes in the presence of virus, and instructions for use.

48. The kit of claim 47, wherein said sample collection mechanisms are chosen from the group consisting of a swab, test tube, syringe, and vial.

49. The kit of claim 47, wherein the reaction chamber is part of a flow-through device, including a housing containing said reaction chamber, wherein said reaction chamber includes a resolving layer having aptamers for a virus and control antigens embedded therein, said housing including a slot for receiving a proximal end of a funnel for applying a sample, a solution of a nanoparticle complexed with a ligand that binds to a virus, and a color developer, and wherein an apparent color of said nanoparticles change in the presence of said virus.

50. The kit of claim 49, further including a vial for collecting a sample, a vial containing a nanoparticle reagent solution, a vial containing a color developer, and a funnel.

51. The kit of claim 47, wherein said virus is chosen from the group consisting of SARS Coronavirus 2

(SARS-CoV-2), adeno-associated virus, aichi virus, Australian bat lyssavirus, BK polyomavirus, banna virus, barmah forest virus, bunyamwera virus, bunyavirus La Crosse, bunyavirus snowshoe hare, cercopithecine herpesvirus, chandipura virus, chikungunya virus, cosavirus A, cowpox virus, coxsackievirus, Crimean-Congo hemorrhagic fever virus, dengue virus, dhori virus, dugbe virus, duvenhage virus, eastern equine encephalitis virus, ebolavirus, echovirus, encephalomyocarditis virus, Epstein-Barr virus, European bat lyssavirus, GB virus C/Hepatitis G virus, hantaan virus, hendra virus, hepatitis A virus, hepatitis B virus, hepatitis C virus, hepatitis E virus, hepatitis delta virus, horsepox virus, human adenovirus, human astrovirus, human coronavirus, human cytomegalovirus, human enterovirus 68, 70, human herpesvirus 1, human herpesvirus 2, human herpesvirus 6, human herpesvirus 7, human herpesvirus 8, human immunodeficiency virus, human papillomavirus 1, human papillomavirus 2, human papillomavirus 16,18, human parainfluenza, human parvovirus B19, human respiratory syncytial virus, human rhinovirus, human SARS coronavirus, human spumaretrovirus, human T-lymphotropic virus, human torovirus, influenza A virus, influenza B virus, influenza C virus, Isfahan virus, JC polyomavirus, Japanese encephalitis virus, Junin arenavirus, Kl polyomavirus, Kunjin virus, Lagos bat virus, Lake Victoria marburgvirus, langat virus, Lassa virus, lordsdale virus, louping ill virus, lymphocytic choriomeningitis virus, machupo virus, mayaro virus, MERS coronavirus, measles virus, mengo encephalomyocarditis virus, merkel cell polyomavirus, mokola virus, molluscum contagiosum virus, monkeypox virus, mumps virus, Murray valley encephalitis virus, New York virus, Nipah virus, Norwalk virus, O'nyong-nyong virus, Orf virus, Oropouche virus, Pichinde virus, poliovirus, punta toro phlebovirus, Puumala virus, rabies virus, Rift valley fever virus, rosavirus A, Ross river virus, rotavirus A, rotavirus B, rotavirus C, rubella virus, Sagiyama virus, Salivirus A, sandfly fever Sicilian virus, Sapporo virus, Semliki forest virus, Seoul virus, simian foamy virus, simian virus 5, Sindbis virus, Southampton virus, St. Louis encephalitis virus, tick- borne powassan virus, Torque teno virus, Toscana virus, Uukuniemi virus, Vaccinia virus, varicella-zoster virus, Variola virus, Venezuelan equine encephalitis virus, vesicular stomatitis virus, western equine encephalitis virus, WU polyomavirus, West Nile virus, Yaba monkey tumor virus, Yaba-like disease virus, yellow fever virus, and Zika virus.

52. The kit of claim 47, wherein said virus is SARS-CoV-2 and said ligand is an aptamer chosen from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, and combinations thereof.

53. A surface test for detecting viruses, comprising: a spray mechanism including a solution of a ligand complexed with a nanoparticle that binds to a virus and changes color in the presence of the virus when sprayed on a surface.

54. The surface test of claim 53, wherein said surface is chosen from the group consisting of tables, desks, doors, door handles, mobile device screens, menus, sinks, and bathrooms.

55. The surface test of claim 53, wherein said virus is chosen from the group consisting of SARS Coronavirus 2 (SARS-CoV-2), adeno-associated virus, aichi virus, Australian bat lyssavirus, BK polyomavirus, banna virus, barmah forest virus, bunyamwera virus, bunyavirus La Crosse, bunyavirus snowshoe hare, cercopithecine herpesvirus, chandipura virus, chikungunya virus, cosavirus A, cowpox virus, coxsackievirus, Crimean-Congo hemorrhagic fever virus, dengue virus, dhori virus, dugbe virus, duvenhage virus, eastern equine encephalitis virus, ebolavirus, echovirus, encephalomyocarditis virus, Epstein-Barr virus, European bat lyssavirus, GB virus C/Hepatitis G virus, hantaan virus, hendra virus, hepatitis A virus, hepatitis B virus, hepatitis C virus, hepatitis E virus, hepatitis delta virus, horsepox virus, human adenovirus, human astrovirus, human coronavirus, human cytomegalovirus, human enterovirus 68, 70, human herpesvirus 1, human herpesvirus 2, human herpesvirus 6, human herpesvirus 7, human herpesvirus 8, human immunodeficiency virus, human papillomavirus 1, human papillomavirus 2, human papillomavirus 16,18, human parainfluenza, human parvovirus B19, human respiratory syncytial virus, human rhinovirus, human SARS coronavirus, human spumaretrovirus, human T-lymphotropic virus, human torovirus, influenza A virus, influenza B virus, influenza C virus, Isfahan virus, JC polyomavirus,

Japanese encephalitis virus, Junin arenavirus, Kl polyomavirus, Kunjin virus, Lagos bat virus, Lake Victoria marburgvirus, langat virus, Lassa virus, lordsdale virus, louping ill virus, lymphocytic choriomeningitis virus, machupo virus, mayaro virus, MERS coronavirus, measles virus, mengo encephalomyocarditis virus, merkel cell polyomavirus, mokola virus, molluscum contagiosum virus, monkeypox virus, mumps virus, Murray valley encephalitis virus, New York virus, Nipah virus, Norwalk virus, O'nyong-nyong virus, Orf virus, Oropouche virus, Pichinde virus, poliovirus, punta toro phlebovirus, Puumala virus, rabies virus, Rift valley fever virus, rosavirus A, Ross river virus, rotavirus A, rotavirus B, rotavirus C, rubella virus, Sagiyama virus, Salivirus A, sandfly fever Sicilian virus, Sapporo virus, Semliki forest virus, Seoul virus, simian foamy virus, simian virus 5, Sindbis virus, Southampton virus, St. Louis encephalitis virus, tick- borne powassan virus, Torque teno virus, Toscana virus, Uukuniemi virus, Vaccinia virus, varicella-zoster virus, Variola virus, Venezuelan equine encephalitis virus, vesicular stomatitis virus, western equine encephalitis virus, WU polyomavirus, West Nile virus, Yaba monkey tumor virus, Yaba-like disease virus, yellow fever virus, and Zika virus.

56. The surface test of claim 53, wherein said virus is SARS-CoV-2 and said ligand is an aptamer chosen from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, and combinations thereof.

57. A surface test for detecting viruses, comprising: a swab for swiping across a surface, wherein said swab is inserted into a reaction chamber containing a solution of at least one aptamer that binds to a virus and changes color in the presence of virus.

58. The surface test of claim 57, wherein said surface is chosen from the group consisting of tables, desks, doors, door handles, mobile device screens, menus, sinks, and bathrooms.

59. The surface test of claim 58, wherein said virus is chosen from the group consisting of SARS Coronavirus 2 (SARS-CoV-2), adeno-associated virus, aichi virus, Australian bat lyssavirus, BK polyomavirus, banna virus, barmah forest virus, bunyamwera virus, bunyavirus La Crosse, bunyavirus snowshoe hare, cercopithecine herpesvirus, chandipura virus, chikungunya virus, cosavirus A, cowpox virus, coxsackievirus, Crimean-Congo hemorrhagic fever virus, dengue virus, dhori virus, dugbe virus, duvenhage virus, eastern equine encephalitis virus, ebolavirus, echovirus, encephalomyocarditis virus, Epstein-Barr virus, European bat lyssavirus, GB virus C/Hepatitis G virus, hantaan virus, hendra virus, hepatitis A virus, hepatitis B virus, hepatitis C virus, hepatitis E virus, hepatitis delta virus, horsepox virus, human adenovirus, human astrovirus, human coronavirus, human cytomegalovirus, human enterovirus 68, 70, human herpesvirus 1, human herpesvirus 2, human herpesvirus 6, human herpesvirus 7, human herpesvirus 8, human immunodeficiency virus, human papillomavirus 1, human papillomavirus 2, human papillomavirus 16,18, human parainfluenza, human parvovirus B19, human respiratory syncytial virus, human rhinovirus, human SARS coronavirus, human spumaretrovirus, human T-lymphotropic virus, human torovirus, influenza A virus, influenza B virus, influenza C virus, Isfahan virus, JC polyomavirus, Japanese encephalitis virus, Junin arenavirus, Kl polyomavirus, Kunjin virus, Lagos bat virus, Lake Victoria marburgvirus, langat virus, Lassa virus, lordsdale virus, louping ill virus, lymphocytic choriomeningitis virus, machupo virus, mayaro virus, MERS coronavirus, measles virus, mengo encephalomyocarditis virus, merkel cell polyomavirus, mokola virus, molluscum contagiosum virus, monkeypox virus, mumps virus, Murray valley encephalitis virus, New York virus, Nipah virus, Norwalk virus, O'nyong-nyong virus, Orf virus, Oropouche virus, Pichinde virus, poliovirus, punta toro phlebovirus, Puumala virus, rabies virus, Rift valley fever virus, rosavirus A, Ross river virus, rotavirus A, rotavirus B, rotavirus C, rubella virus, Sagiyama virus, Salivirus A, sandfly fever Sicilian virus, Sapporo virus, Semliki forest virus, Seoul virus, simian foamy virus, simian virus 5, Sindbis virus, Southampton virus, St. Louis encephalitis virus, tick- borne powassan virus, Torque teno virus, Toscana virus, Uukuniemi virus, Vaccinia virus, varicella-zoster virus, Variola virus, Venezuelan equine encephalitis virus, vesicular stomatitis virus, western equine encephalitis virus, WU polyomavirus, West Nile virus, Yaba monkey tumor virus, Yaba-like disease virus, yellow fever virus, and Zika virus.

60. The surface test of claim 58, wherein said virus is SARS-CoV-2 and said ligand is an aptamer chosen from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, and combinations thereof.

61. A method of detecting viruses on surfaces, including the steps of: spraying a solution of a nanoparticle complexed with a ligand that binds to a virus, wherein the nanoparticle's apparent color changes in the presence of virus on a surface; and detecting a color change on the surface indicating the presence of virus.

62. The method of claim 61, wherein the surface is chosen from the group consisting of tables, desks, doors, door handles, mobile device screens, menus, sinks, and bathrooms.

63. The method of claim 61, wherein the virus is chosen from the group consisting of SARS

Coronavirus 2 (SARS-CoV-2), adeno-associated virus, aichi virus, Australian bat lyssavirus, BK polyomavirus, banna virus, barmah forest virus, bunyamwera virus, bunyavirus La Crosse, bunyavirus snowshoe hare, cercopithecine herpesvirus, chandipura virus, chikungunya virus, cosavirus A, cowpox virus, coxsackievirus, Crimean-Congo hemorrhagic fever virus, dengue virus, dhori virus, dugbe virus, duvenhage virus, eastern equine encephalitis virus, ebolavirus, echovirus, encephalomyocarditis virus, Epstein-Barr virus, European bat lyssavirus, GB virus C/Hepatitis G virus, hantaan virus, hendra virus, hepatitis A virus, hepatitis B virus, hepatitis C virus, hepatitis E virus, hepatitis delta virus, horsepox virus, human adenovirus, human astrovirus, human coronavirus, human cytomegalovirus, human enterovirus 68, 70, human herpesvirus 1, human herpesvirus 2, human herpesvirus 6, human herpesvirus 7, human herpesvirus 8, human immunodeficiency virus, human papillomavirus 1, human papillomavirus 2, human papillomavirus 16,18, human parainfluenza, human parvovirus B19, human respiratory syncytial virus, human rhinovirus, human SARS coronavirus, human spumaretrovirus, human T-lymphotropic virus, human torovirus, influenza A virus, influenza B virus, influenza C virus, Isfahan virus, JC polyomavirus,

64. The method of claim 61, wherein said virus is SARS-CoV-2 and said ligand is an aptamer chosen from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO:

5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, and combinations thereof.