Multi-analyte rapid test system

Abstract

The technology relates in part to multi-analyte rapid tests. In certain aspects, the technology relates in part to interchangeable multi-analyte rapid tests. In certain aspects, the technology relates in part to enzyme-based multi-analyte rapid tests. In certain aspects, the technology relates to enzyme-immobilized-based multi-analyte rapid tests. In certain aspects, the technology relates to interchangeable multi-analyte rapid tests. In certain aspects, the technology relates to enzyme-based interchangeable multi-analyte rapid tests. In certain aspects, the technology relates to enzyme-based multi-analyte lateral flow rapid tests. In certain aspects, the technology relates to enzyme-immobilized-based interchangeable multi-analyte lateral flow rapid tests.

Description

[0001] PATENT

[0002] AIPA-1039PCT2

[0003] MULTI-ANALYTE RAPID TEST SYSTEM

[0004] Field

[0005] The technology relates in part to multi-analyte rapid tests. In certain aspects, the technology relates in part to interchangeable multi-analyte rapid tests. In certain aspects, the technology relates in part to enzyme-based multi-analyte rapid tests. In certain aspects, the technology relates to enzyme-immobilized-based multi-analyte rapid tests. In certain aspects, the technology relates to interchangeable multi-analyte rapid tests. In certain aspects, the technology relates to enzymebased interchangeable multi-analyte rapid tests. In certain aspects, the technology relates to enzyme-based multi-analyte lateral flow rapid tests. In certain aspects, the technology relates to enzyme-immobilized-based interchangeable multi-analyte lateral flow rapid tests.

[0006] Background

[0007] Rapid detection systems are increasingly important in fields such as food safety, medical diagnostics, and environmental monitoring. Existing detection technologies often require long reaction times, specific environmental conditions, or complex equipment, which limits their applicability in field and point-of-care settings. There is a need for a versatile, portable, and scalable platform capable of detecting multiple analytes in liquid samples with immediate visual or instrument-readable results.

[0008] Summary

[0009] Provided herein in certain aspects are rapid detection platforms comprising a) a substrate carrier with one or more interchangeable reaction zones specific to one or more analytes, and b) a detection system, where the platform is capable of detecting the one or more analytes in a liquid sample and produces a signal upon interaction with the one or more analytes.

[0010] Also provided in certain aspects are methods for detecting a presence or absence of a plurality of analyte species in a liquid sample using a rapid detection platform provided herein, comprising i) contacting the liquid sample with the substrate carrier; ii) allowing the sample to flow through the substrate carrier; and iii) detecting the presence or absence of the plurality of analyte species according to a plurality of corresponding signals.

[0011] Also provided in certain aspects are rapid detection platforms comprising a substrate carrier with immobilized enzymes, a chromogenic or fluorogenic detection system, and an interchangeable reaction mechanism capable of detecting one or more analytes selected from sugars, proteins, toxins, and small molecules in a liquid sample, where a color or fluorescence change occurs upon interaction with the target analyte, yielding immediate visual or instrument-readable results.PATENT

[0012] AIPA-1039PCT2

[0013] Also provided in certain aspects are rapid detection platforms comprising a) a substrate carrier with immobilized enzymes specific to one or more analytes; b) a chromogenic or fluorogenic detection system; and c) an interchangeable reaction mechanism; where the platform detects analytes selected from sugars, proteins, toxins, or small molecules in a liquid sample and produces an immediate visual or instrument-readable signal upon interaction with the analyte.

[0014] Also provided in certain aspects are universal rapid detection platforms comprising a) a substrate carrier with immobilized enzymes specific to one or more analytes; b) a chromogenic or fluorogenic detection system; and c) an interchangeable reaction mechanism; where the platform detects analytes selected from sugars, proteins, toxins, small molecules, or other biological or chemical substances in a liquid sample and produces an immediate visual or instrument-readable signal upon interaction with the analyte.

[0015] Also provided in certain aspects are methods for detecting an analyte in a liquid sample using a rapid detection platform provided herein, comprising the steps of: i) applying the liquid sample to the substrate carrier; ii) allowing the sample to flow through immobilized enzymes; and iii) observing a color or fluorescence change indicating the presence of the analyte.

[0016] Also provided in certain aspects are general-purpose detection systems comprising a rapid detection platform provided herein, where the platform can be used for food safety, medical diagnostics, environmental monitoring, and / or industrial quality control.

[0017] Certain implementations are described further in the following description, examples and claims, and in the drawings.

[0018] Brief Description of the Drawings

[0019] The drawings illustrate certain implementations of the technology and are not limiting. For clarity and ease of illustration, the drawings are not made to scale and, in some instances, various aspects may be shown exaggerated or enlarged to facilitate an understanding of particular implementations.

[0020] Figs. 1 A and 1 B show an example lateral flow test system with three interchangeable reaction zone modules (A, B, C) that assemble in a serial configuration along the test strip flow axis.

[0021] Figs. 2A and 2B show an example lateral flow test system with two interchangeable reaction zone modules (A, B) that assemble a serial configuration along the test strip flow axis.

[0022] Figs. 3A and 3B show an example lateral flow test system with three interchangeable reaction zone modules (A, B, C) that assemble in a parallel configuration on the test strip.PATENT

[0023] AIPA-1039PCT2

[0024] Figs. 4A and 4B show an example lateral flow test system with two interchangeable reaction zone modules (A, B) that assemble in a parallel configuration on the test strip.

[0025] Fig. 5 shows examples of reaction zone configurations in a reaction zone module.

[0026] Provided in the following table is a listing of elements identified in Figures 1-5:

[0027]

[0028] PATENT

[0029] AIPA-1039PCT2

[0030]

[0031] PATENT

[0032] AIPA-1039PCT2

[0033] Detailed Description

[0034] Provided herein are rapid detection platforms comprising a substrate carrier with immobilized enzymes, a chromogenic or fluorogenic detection system, and an interchangeable reaction mechanism. The platforms are capable of detecting one or more analytes (e.g., sugars, proteins, toxins, and / or small molecules) in a liquid sample. The system produces an immediate color or fluorescence change upon interaction with the target analyte, enabling visual interpretation or quantitative analysis via instrumentation. Advantages of the rapid detection platforms provided herein include, for example, versatility (e.g., capable of detecting multiple analytes using the same platform by changing immobilized enzymes and reagents), rapid results (e.g., provides immediate feedback without the need for incubation or specialized equipment), portability (e.g., compact and lightweight, suitable for field use), scalability (e.g., easily adaptable for large-scale manufacturing and customization), and cost-effectiveness (e.g., uses inexpensive materials and reagents, making it accessible for widespread applications). Accordingly, the rapid detection platforms provided herein offer a versatile, rapid, and scalable solution for multi-analyte detection, addressing critical needs in diagnostics, quality control, and monitoring. The rapid detection platforms provided herein demonstrate significant improvements in usability, speed, and adaptability compared to existing detection systems.

[0035] Platform design

[0036] Rapid detection platforms provided herein generally include a substrate carrier, immobilized enzymes, a detection system, and an interchangeable reaction mechanism. In certain configurations, a rapid detection platform comprises a test sample receiving region (also referred to as sample pad or sample application pad), a test region with capacity for interchangeable reaction zone modules, a control region, and a reservoir region (also referred to as an absorption pad). In some embodiments, a rapid detection platform is a lateral flow test device comprising a lateral flow test strip. A test strip may comprise comprises a dry matrix material capable of transporting a liquid along a flow axis by capillarity with a start zone for receiving a sample and a reaction zone in which a reaction (e.g., enzyme reaction) takes place which is specific for an analyte in the sample and which generates a detectable signal. In some embodiments, a rapid detection platform herein further comprises a portable housing unit for field or point-of-care use, with integrated sample application and interpretation zones. In some embodiments, a rapid detection platform herein is adapted for simultaneous detection of multiple analytes using spatially separated reaction zones. As an example, a lateral flow test herein may operate as follows. Upon application of a liquid sample, fluid flows from a sample pad to a substrate carrier on a lateral flow test strip. As the sample progresses from the sample pad to the test strip, it is driven through the strip by capillaryPATENT

[0037] AIPA-1039PCT2

[0038] action in the substrate carrier. This action facilitates selective interactions between the analyte and the analyte-detecting reagent, such as enzymatic interactions, at designated test lines or test zones and at control lines, which may include immobilized enzymes, antibodies, antigens or other analytedetecting reagents. The lateral flow capillary action wicks the sample through the test strip, where analyte interactions with an analyte-detecting reagent, such as an immobilized enzyme, occur with the target analyte in the sample, resulting in the presence of a control line and the presence or absence of a test line forming on the test strip. These interactions lead to the visualization of a line at the test site if the target analyte is present, while the control line binds or interacts with other analytes, reagents, sample components, or enzymatic products to confirm the assay's functionality. The detection range (concentration of analyte) depends on the nature of the analyte, example nonlimiting ranges being from about the 0.5 ng / ml to about 50 ng / ml with visual detection and about 5 pg / ml to about 50 ng / ml using a reader.

[0039] Sample receiving region

[0040] A rapid detection platform provided herein may comprise a sample receiving region. A sample receiving region typically is situated at one end of the test strip (sometimes referred to herein at the proximal end). The sample to be analyzed is applied to the sample receiving region. In some embodiments, a sample receiving region comprises a sample pad composed of an absorbent paper. Glass fiber materials, fibrous plastic materials such as Porex® sheet materials (Porex Corporation), and non-woven fabrics comprising such materials as viscose and polyester may also be used. Such materials may be in a single sheet or layer. The volume capacity of the sample pad generally defines the initial sample uptake of the device. The sample pad also may act as a filter to help to prevent unwanted particulate materials from reaching the reaction zone(s). The material of the sample pad also may help to control the release of any impregnated mobilizable components of the assay to the rest of the device.

[0041] Liquid samples may be added to a sample receiving region as single drops e.g. by using a pipette or by dipping the sample receiving region in a liquid sample. A sample receiving region can comprise components which are dried onto it such as components of an enzyme assay and / or chemicals to prevent (“block”) non-specific binding effects at the reaction zone and / or to chemicals to enhance hydrophilic properties, rehydration of assay components and lateral flow characteristics and / or analyte release agents and / or extractants (e.g., detergent extractants).

[0042] Substrate carrier

[0043] A rapid detection platform provided herein may comprise a substrate carrier. A substrate carrier also may be referred herein to as a test strip, a lateral flow test strip, or a lateral flow strip. A substrate carrier herein may contain any suitable material for conducting liquid flow. In somePATENT

[0044] AIPA-1039PCT2

[0045] embodiments, a substrate carrier comprises an absorbent material which may include polymer fibers, copolymer fibers, sponge or foam materials, cotton fiber materials, blended fibers such as, for example, a blend of polyethylene and polypropylene and combinations thereof including, but not limited to, rayons, polyesters, polyethylene, polypropylene, nylons, polyamides, carbon fibers, alginates, cottons, silks, dacron, rayon, polyurethane, and combinations thereof. In some embodiments, a substrate carrier comprises filter paper, nitrocellulose membranes, polymer-based films, or porous wood materials capable of facilitating capillary action for liquid flow. In some embodiments, a substrate carrier is chemically functionalized (e.g., with carboxyl or amine groups) to enhance the immobilization of enzymes and / or reagents. In some embodiments, a substrate carrier is treated or modified to enhance durability and compatibility with organic solvents, aqueous solutions, or high-sensitivity analytes.

[0046] In some configurations, a substrate carrier comprises a matrix material fixed to a solid support in form of one or more absorbent interconnecting porous pads. Typically, matrix material is compatible with the stability of the relevant assay reagents, the chemistry of the assay, and with required lateral flow characteristics such as flow rate. A solid support is typically non-water absorbent material, often plastic such as polyester sheet. The support is typically bonded to the dry matrix materials by a suitable adhesive.

[0047] Reaction zone

[0048] A rapid detection platform provided herein may comprise one or more reaction zones. A reaction zone also may be referred to as a test zone. A reaction zone may be located directly adjacent to a sample pad or may be located further down the test strip. In some embodiments, a reaction zone contains one or more components of an enzyme assay (e.g., in a stable state). In some embodiments, a reaction zone contains one or more enzymes (i.e., enzyme species) immobilized to the reaction zone. Additional components may include additional immobilized enzymes and / or immobilized or dried enzyme substrates, co-substrates co-factors, and the like. A reaction zone may be a pad or a membrane suitable for lateral flow and for non-covalently or covalently immobilizing protein without denaturation. In some embodiments a reaction zone is composed of a nitrocellulose membrane (e.g., with a high protein binding capacity). Such membrane may also incorporate cellulose acetate. The pad or membrane may be treated before and / or after enzyme immobilization in order to minimize unwanted effects such as non-specific binding of analyte and / or assay components and / or in order to enhance hydrophilic properties and lateral flow. Typically, it is on the reaction zone that the reactions comprising the assay occur and the product indicating the presence of the analyte in the original sample is formed. In some embodiments, multiple reaction zones can be incorporated into zone designs as required and / or multiple enzyme components canPATENT

[0049] AIPA-1039PCT2

[0050] be deposited at different points on the reaction zone. In some embodiments, more than one enzyme or group of enzymes or other reagents capable of detecting more than one analyte of interest at separate locations may be incorporated within the test strip. In some embodiments, both enzyme assay and immunoassay are incorporated within the same test strip.

[0051] In certain configurations, a test device further comprises a film made of plastic or other suitable material overlaying the reaction zone(s) of the device to prevent contamination of the device during operation while still allowing any signal produced by the device to be detected. The employment of this film allows easy handling and operation without the risk of contamination.

[0052] Interchangeable reaction mechanism

[0053] A rapid detection platform provided herein may comprise an interchangeable reaction mechanism. In such configurations, the platform allows for swapping out reaction modules to detect different analytes by embedding modular reaction zones or cartridges within the substrate or test strip. The interchangeable reagent mechanism allows modular customization for detecting multiple analytes, including sugars, proteins, toxins, pH levels, ions, and / or other substances. An interchangeable reaction mechanism may include 2 or more reaction modules. For example, an interchangeable reaction mechanism may include 2, 3, 4, 5, 6, 7, 8, 9, 10, or more reaction modules. In some embodiments, the reaction modules are configured to assemble in a serial configuration along the test strip flow axis (e.g., see Figs 1 A, 1 B, 2A, 2B). In some embodiments, the reaction modules are configured to assemble in a parallel configuration (e.g., see Figs 3A, 3B, 4A, 4B). In some embodiments, the reaction modules are configured to assemble in a parallel configuration and in a serial configuration along the test strip flow axis.

[0054] Reaction modules may comprise one or more immobilized enzymes distributed throughout the surface of the module 100, 300. In some configurations, reaction modules comprise one or more immobilized enzymes distributed within a specific region (test zone 210, 410) of the module 200, 400. Reaction modules may be assembled such that they abut directly against one another and / or other regions of the test strip (e.g., sample pad, control zone, absorbent pad). Reaction modules may be assembled such that part of a module overlaps with another module and / or other regions of the test strip (e.g., sample pad, control zone, absorbent pad). In some embodiments, reaction modules may be configured to fit or snap together with each other and / or with other regions of the test strip (e.g., sample pad, control zone, absorbent pad) like jigsaw puzzle pieces. In some embodiments, an external housing unit facilitates installation and fit of the reaction modules.

[0055] Figures 1-4 show example rapid detection platform systems with various interchangeable reaction mechanism configurations. A particular non-limiting implementation of a rapid detection platform is system 10 illustrated in Fig. 1 A. System 10 includes lateral flow test strip base 18, test samplePATENT

[0056] AIPA-1039PCT2

[0057] receiving region 11 at the proximal end of test strip 18, which also may be referred to as a sample pad or sample application pad, test region 12 with capacity for three interchangeable reaction zone modules that assemble in a serial configuration along the test strip flow axis, control region 13, reservoir region 14 at the distal end of test strip 18, also referred to as an absorption pad, uninstalled reaction zone module A 15, uninstalled reaction zone module B 16, and uninstalled reaction zone module C 17. System 10 as assembled is system 20 that includes installed reaction zone module A 25, installed reaction zone module B 26, and installed reaction zone module C 27. Another non-limiting implementation of a rapid detection platform is system 30 illustrated in Fig. 2A. System 30 includes lateral flow test strip base 37, test sample receiving region 31 at the proximal end of test strip 37, which also may be referred to as a sample pad or sample application pad, test region 32 with capacity for two interchangeable reaction zone modules that assemble in a serial configuration along the test strip flow axis, control region 33, reservoir region 34 at the distal end of test strip 37, also referred to as an absorption pad, uninstalled reaction zone module A 35, and uninstalled reaction zone module B 66. System 30 as assembled is system 40 that includes installed reaction zone module A 45 and installed reaction zone module B 46.

[0058] Another non-limiting implementation of a rapid detection platform is system 50 illustrated in Fig. 3A. System 50 includes lateral flow test strip base 58, test sample receiving region 51 at the proximal end of test strip 58, which also may be referred to as a sample pad or sample application pad, test region 52 with capacity for three interchangeable reaction zone modules that assemble in a parallel configuration, control region 53, reservoir region 54 at the distal end of test strip 58, also referred to as an absorption pad, uninstalled reaction zone module A 55, uninstalled reaction zone module B 56, and uninstalled reaction zone module C 57. System 50 as assembled is system 60 that includes installed reaction zone module A 65, installed reaction zone module B 66, and installed reaction zone module C 67.

[0059] Another non-limiting implementation of a rapid detection platform is system 70 illustrated in Fig. 4A. System 70 includes lateral flow test strip base 77, test sample receiving region 71 at the proximal end of test strip 77, which also may be referred to as a sample pad or sample application pad, test region 72 with capacity for two interchangeable reaction zone modules that assemble in a parallel configuration, control region 73, reservoir region 74 at the distal end of test strip 77, also referred to as an absorption pad, uninstalled reaction zone module A 75, and uninstalled reaction zone module B 76. System 70 as assembled is system 80 that includes installed reaction zone module A 85 and installed reaction zone module B 86.PATENT

[0060] AIPA-1039PCT2

[0061] Immobilized enzymes

[0062] A rapid detection platform provided herein may comprise one or more immobilized enzymes.

[0063] Enzymes specific to target analytes are immobilized onto the substrate carrier using techniques such as covalent binding, adsorption, and / or entrapment in polymer matrices. In some embodiments, an analyte is detected according to its transient participation in an enzyme reaction. During this enzyme reaction an analyte is chemically modified and thus directly or indirectly participates in a reaction that creates a detectable signal. As a consequence, specificity for an analyte is provided by an enzyme which uses the analyte as a substrate, co-substrate or co-factor and chemically modifies it. The signal for detection and measurement of the analyte is thus dependent upon the turnover of the enzyme-mediated reaction involving the analyte.

[0064] Examples of enzymes include lactase (for lactose detection), which hydrolyzes lactose into glucose and galactose; proteases (for protein detection), which break down proteins into peptides and / or amino acids; oxidases (e.g., glucose oxidase or galactose oxidase), which convert analyte-derived products into detectable intermediates.

[0065] Other non-limiting examples of enzyme assays that may be adapted for use in a rapid detection platform herein include the detection of glucose by the enzymes glucose dehydrogenase and diaphorase using a substrate formulation incorporating NAD and a suitable diaphorase substrate, for example NBT; the detection of urea in biological fluids by the enzymes glutamate dehydrogenase, diaphorase and urease using a substrate formulation incorporating NADH, 2-oxoglutarate and a suitable diaphorase substrate, for example NBT; the detection of serum cholesterol by the enzymes cholesterol oxidase and peroxidase using a substrate formulation incorporating suitable peroxidase substrates, for example 4-aminophenazone and phenol; the detection of alcohol by the enzymes alcohol dehydrogenase and diaphorase using a substrate formulation incorporating NAD and a suitable diaphorase substrate, for example NBT; the detection of carbon dioxide and / or bicarbonate in biological fluids by the enzymes phosphoenolpyruvate carboxylase, malate dehydrogenase and diaphorase using a substrate formulation incorporating phosphoenolpyruvate, oxaloacetate, NADH and a suitable diaphorase substrate, for example NBT; and the detection of biomass using a NAD(H) calorimetric assay

[0066] Detection system

[0067] A rapid detection platform provided herein may comprise a detection system. A detection system may be a chromogenic detection system or a fluorogenic detection system. Chromogenic substrates may include, for example, tetramethylbenzidine (TMB), 3,3'-diaminobenzidine (DAB), or ABTS, which change color in response to enzymatic reactions. Fluorogenic substrates (e.g., AMPLEX Red, resorufin-based dyes) produce fluorescence when oxidized or reduced. In somePATENT

[0068] AIPA-1039PCT2

[0069] embodiments, a signal is interpreted visually, using a compatible reader device, or through integration with electronic sensing components.

[0070] In one example, lactose may be detected according to an enzymatic reaction that includes lactase which hydrolyzes lactose into glucose and galactose, oxidase converts galactose into hydrogen peroxide, and hydrogen peroxide reacts with HRP and TMB, yielding a blue color. In another example, proteins may be detected according to an enzymatic reaction that includes a protease which hydrolyzes proteins into peptides or amino acids, and amino acids interact with ninhydrin or other reagents to produce a visible color. In another example, toxins may be detected according to an enzymatic reaction that includes a specific enzyme or aptamer that binds the toxin, triggering a cascade that produces a detectable signal (e.g., fluorescence or color change).

[0071] Reservoir region

[0072] A rapid detection platform provided herein may comprise a reservoir region or absorbent pad. A reservoir region generally draws liquid sample from the matrix of the test strip through the device and therefore drives the capillary flow from the sample pad. Flow of solution into the reservoir region ensures a sustained flow from the sample pad across the reaction zone(s) of the immobilized enzymes to maximize the amount of product formed. The reservoir region is typically composed of materials similar to those employed for the sample pad. In some embodiments, the reservoir region is composed of an absorbent paper. An assay typically reaches endpoint when the volume capacity of the absorbent pad is filled, assuming that sample volume is not limiting.

[0073] In certain configurations, the various absorbent zones of the rapid detection platform provided herein are all in the same plane, allowing capillary flow of a liquid sample between the zones. In the process, any component(s) deployed in dried state on the test strip, such as detergent to extract the analyte from matrices within the sample, are reconstituted into solution. Certain zones may contain further reagents like stabilizing agents or buffers (e.g., to support storage of certain components of an enzyme reaction or to provide reaction conditions (e.g. pH) that are suitable for an enzyme reaction). For some applications, an enzyme reaction may be combined with a chemical reaction (e.g., to improve the generation of a detectable signal). In certain configurations, components of a chemical reaction are also included in dry state in certain zones of the test strip.

[0074] Housing

[0075] A rapid detection platform provided herein may comprise a test device housing (e.g., a housing containing a test strip, a housing containing a substrate carrier with interchangeable reaction zones). A housing may provide additional strength and rigidity to the platform and to facilitate handling of the platform without contamination. In certain configurations, a rapid detection platformPATENT

[0076] AIPA-1039PCT2

[0077] provided herein comprises a hollow test device housing that includes a proximal terminus, a distal terminus, a base, a cover, sidewalls between the proximal terminus and distal terminus, a sample collection reservoir at the proximal terminus, and a window for viewing test results. In some embodiments, a housing base comprises one or more regions for supporting interchangeable reaction zones. Typically, a test strip is only partially covered by the housing. For example, a sample pad may not be covered by the housing to ensure easy application of a sample. A test devise housing may be made of any suitable material, polymer or polymer mixture. Non-limiting examples of polymers include polyethylene (PE), low density polyethylene (LDPE), high-density polyethylene (HDPE), polypropylene (PP), high impact polystyrene (HIPS), polyvinyl chloride (PVC), polyethylene terephthalate (PET), amorphous polyethylene terephthalate (APET), acrylonitrile butadiene styrene (ABS), and polycarbonate (PC).

[0078] Method of detection

[0079] Provided herein are methods of detecting a presence or absence of a plurality of analyte species in a liquid sample using a rapid detection platform described herein. Generally, a method of detection includes i) contacting a liquid sample with a substrate carrier; ii) allowing the sample to flow through the substrate carrier; and iii) detecting the presence or absence of a plurality of analyte species according to a plurality of corresponding signals. In some embodiments, a liquid sample is applied directly to a substrate carrier, initiating the flow of the sample through the test strip. In some embodiments, a liquid sample is applied directly to a sample pad in contact with a substrate carrier, initiating the flow of the sample through the test strip. In some embodiments, a target analyte reacts with the immobilized enzyme(s), producing specific products (e.g., glucose, amino acids, or hydrogen peroxide). In some embodiments, enzymatic products interact with a chromogenic detection system or fluorogenic detection system to produce an immediate and detectable signal. Results may be interpreted visually (e.g., color intensity) or via a compatible reader device (e.g., fluorescence or absorbance spectrophotometer).

[0080] Industrial applications

[0081] A rapid detection platform provided herein may be useful for a variety of industrial applications. For example, the food safety industry may use the rapid detection platform provided herein for detecting lactose, allergens, and / or contaminants in dairy products and beverages. A rapid detection platform provided herein may be useful for medical diagnostics, for example, identifying specific biomarkers in blood, urine, and / or saliva samples. A rapid detection platform provided herein may be useful for environmental monitoring, for example, measuring toxins and / or pollutants in water samples. A rapid detection platform provided herein may be useful for pharmaceutical quality control, for example, detecting impurities and / or verifying ingredient concentrations.PATENT

[0082] AIPA-1039PCT2

[0083] Certain Implementations

[0084] Following are non-limiting examples of certain implementations of the technology.

[0085] A1 . A rapid detection platform comprising a substrate carrier with immobilized enzymes, a chromogenic or fluorogenic detection system, and an interchangeable reaction mechanism capable of detecting one or more analytes selected from sugars, proteins, toxins, and small molecules in a liquid sample, wherein a color or fluorescence change occurs upon interaction with the target analyte, yielding immediate visual or instrument-readable results.

[0086] B1 . A universal rapid detection platform comprising:

[0087] a) a substrate carrier with immobilized enzymes specific to one or more analytes;

[0088] b) a chromogenic or fluorogenic detection system; and

[0089] c) an interchangeable reaction mechanism;

[0090] wherein the platform detects analytes selected from sugars, proteins, toxins, small molecules, or other biological or chemical substances in a liquid sample and produces an immediate visual or instrument-readable signal upon interaction with the analyte.

[0091] B2. The platform of embodiment B2, wherein the substrate carrier is selected from filter paper, nitrocellulose membranes, polymer-based films, porous wood materials, or other solid materials capable of facilitating liquid flow or reagent retention.

[0092] B3. The platform of embodiment B1 or B2, wherein the chromogenic detection system includes tetramethylbenzidine (TMB), ABTS, 3,3'-diaminobenzidine (DAB), or other reagents producing visible color changes.

[0093] B4. The platform of embodiment B1 or B2, wherein the fluorogenic detection system includes AMPLEX Red, resorufin-based dyes, or other fluorescence-producing reagents.

[0094] B5. The platform any one of embodiments B1-B4, wherein the enzymes include lactase, glucose oxidase, galactose oxidase, proteases, or other analyte-specific enzymes.

[0095] B6. The platform of any one of embodiments B1-B5, wherein the interchangeable reaction mechanism allows modular customization for detecting multiple analytes, including sugars, proteins, toxins, pH levels, ions, or other substances.

[0096] B7. The platform of any one of embodiments B1-B6, wherein the signal is interpreted visually, using a compatible reader device, or through integration with electronic sensing components.

[0097] B8. The platform of any one of embodiments B1-B7, further comprising a portable housing unit for field or point-of-care use, with integrated sample application and interpretation zones.PATENT

[0098] AIPA-1039PCT2

[0099] B9. The platform of any one of embodiments B1-B8, wherein the substrate carrier is treated or modified to enhance durability and compatibility with organic solvents, aqueous solutions, or high-sensitivity analytes.

[0100] B10. A method for detecting an analyte in a liquid sample using the platform of any one of embodiments B1-B9, comprising the steps of:

[0101] i) applying the liquid sample to the substrate carrier;

[0102] ii) allowing the sample to flow through immobilized enzymes; and

[0103] iii) observing a color or fluorescence change indicating the presence of the analyte.

[0104] B11 . The method of embodiment B9, wherein the platform is adapted for simultaneous detection of multiple analytes using spatially separated reaction zones.

[0105] B12. A general-purpose detection system, as in any one of embodiments B1-B9, wherein the platform can be used for food safety, medical diagnostics, environmental monitoring, and / or industrial quality control.

[0106] C1 . A rapid detection platform comprising:

[0107] a) a substrate carrier with immobilized enzymes specific to one or more analytes;

[0108] b) a chromogenic or fluorogenic detection system; and

[0109] c) an interchangeable reaction mechanism;

[0110] wherein the platform detects analytes selected from sugars, proteins, toxins, or small molecules in a liquid sample and produces an immediate visual or instrument-readable signal upon interaction with the analyte.

[0111] C2. The platform of embodiment C1 , wherein the substrate carrier is selected from filter paper, nitrocellulose membranes, polymer-based films, or porous wood materials.

[0112] C3. The platform of embodiment C1 or C2, wherein the chromogenic detection system includes tetramethylbenzidine (TMB), ABTS, or 3,3'-diaminobenzidine (DAB).

[0113] C4. The platform of embodiment C1 or C2, wherein the fluorogenic detection system includes AMPLEX Red or resorufin-based dyes.

[0114] C5. The platform of any one of embodiments C1-C4, wherein the enzymes include lactase, glucose oxidase, galactose oxidase, or proteases.

[0115] C6. The platform of any one of embodiments C1-C5, wherein the interchangeable reaction mechanism allows modular customization for detecting different analytes.

[0116] C7. The platform of any one of embodiments C1-C6, wherein the signal is interpreted visually or using a compatible reader device.PATENT

[0117] AIPA-1039PCT2

[0118] C8. A method for detecting an analyte in a liquid sample using the platform of any one of embodiments C1-C7, comprising the steps of:

[0119] i) applying the liquid sample to the substrate carrier;

[0120] ii) allowing the sample to flow through immobilized enzymes; and

[0121] iii) observing a color or fluorescence change indicating the presence of the analyte.

[0122] Examples

[0123] The examples set forth below illustrate certain implementations and do not limit the technology. Example 1: Rapid test platform for detecting lactose

[0124] An example configuration of the platform includes:

[0125] Material: Nitrocellulose membrane or porous wood material as the substrate carrier.

[0126] Substrate carrier configuration: Interchangeable reaction zones.

[0127] Immobilized enzymes: Lactase, glucose oxidase, and horseradish peroxidase.

[0128] Chromogenic substrate: Tetramethylbenzidine (TMB).

[0129] Sample: Milk or a lactose-containing beverage.

[0130] Procedure: Apply a drop of the sample onto the test strip. Observe the development of a blue color within 30 seconds, indicating the presence of lactose.

[0131] The entirety of each patent, patent application, publication and document referenced herein is incorporated by reference. Citation of patents, patent applications, publications and documents is not an admission that any of the foregoing is pertinent prior art, nor does it constitute any admission as to the contents or date of these publications or documents. Their citation is not an indication of a search for relevant disclosures. All statements regarding the date(s) or contents of the documents is based on available information and is not an admission as to their accuracy or correctness. The technology has been described with reference to specific implementations. The terms and expressions that have been utilized herein to describe the technology are descriptive and not necessarily limiting. Certain modifications made to the disclosed implementations can be considered within the scope of the technology. Certain aspects of the disclosed implementations suitably may be practiced in the presence or absence of certain elements not specifically disclosed herein.PATENT

[0132] AIPA-1039PCT2

[0133] Each of the terms “comprising,” “consisting essentially of,” and “consisting of” may be replaced with either of the other two terms. The term “a” or “an” can refer to one of or a plurality of the elements it modifies (e.g., “a reagent” can mean one or more reagents) unless it is contextually clear either one of the elements or more than one of the elements is described. The term “about” as used herein refers to a value within 10% of the underlying parameter (i.e., plus or minus 10%; e.g., a weight of “about 100 grams” can include a weight between 90 grams and 110 grams). Use of the term “about” at the beginning of a listing of values modifies each of the values (e.g., “about 1 , 2 and 3” refers to "about 1 , about 2 and about 3"). When a listing of values is described the listing includes all intermediate values and all fractional values thereof (e.g., the listing of values "80%, 85% or 90%" includes the intermediate value 86% and the fractional value 86.4%). When a listing of values is followed by the term "or more," the term "or more" applies to each of the values listed (e.g., the listing of "80%, 90%, 95%, or more" or "80%, 90%, 95% or more" or "80%, 90%, or 95% or more" refers to "80% or more, 90% or more, or 95% or more"). When a listing of values is described, the listing includes all ranges between any two of the values listed (e.g., the listing of "80%, 90% or 95%" includes ranges of "80% to 90%, " "80% to 95%" and "90% to 95%").

[0134] Certain implementations of the technology are set forth in the claim(s) that follow(s).

Claims

PATENTAIPA-1039PCT2What is claimed is:

1. A rapid detection platform comprising:a) a substrate carrier with one or more interchangeable reaction zones specific to one or more analytes, andb) a detection system,wherein the platform is capable of detecting the one or more analytes in a liquid sample and produces a signal upon interaction with the one or more analytes.

2. The rapid detection platform of claim 1 , wherein the substrate carrier comprises two or more interchangeable reaction zones.

3. The rapid detection platform of claim 1 , wherein the substrate carrier comprises three or more interchangeable reaction zones.

4. The rapid detection platform of any one of claims 1-3, wherein the reaction zones comprise immobilized enzymes specific to the corresponding one or more analytes.

5. The rapid detection platform of claim 4, wherein the enzymes are chosen from one or more of lactase, glucose oxidase, and galactose oxidase.

6. The rapid detection platform of claim 4 or 5, wherein the enzymes are chosen from one or more proteases.

7. The rapid detection platform of any one of claims 1-6, wherein the detection system comprises a chromogenic detection system.

8. The rapid detection platform of any one of claims 1-7, wherein the detection system comprises a fluorogenic detection system.

9. The rapid detection platform of any one of claims 1-8, wherein the signal is a visual signal.

10. The rapid detection platform of any one of claims 1-9, wherein the signal is an instrument-readable signal.11 . The rapid detection platform of any one of claims 1 -10, wherein the one or more analytes are chosen from one or more biological analytes.

12. The rapid detection platform of any one of claims 1-11 , wherein the one or more analytes are chosen from one or more chemical analytes.

13. The rapid detection platform of any one of claims 1-12, wherein the one or more analytes are chosen from one or more of sugars, proteins, toxins, and small molecules.

14. The rapid detection platform of any one of claims 1-13, wherein the substrate carrier is chosen from filter paper, nitrocellulose membranes, polymer-based films, and porous wood materials.PATENTAIPA-1039PCT215. A method for detecting a presence or absence of a plurality of analyte species in a liquid sample using the rapid detection platform of any one of claims 1-14, comprising:i) contacting the liquid sample with the substrate carrier;ii) allowing the sample to flow through the substrate carrier; andiii) detecting the presence or absence of the plurality of analyte species according to a plurality of corresponding signals.

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