Flow Analyzer

Abstract

A lateral flow high throughput assay device analyzer for preparing and analyzing multiple lateral flow assay samples is described. The analyzer includes a cartridge stage for supporting an assay cartridge, a height adjustment mechanism, and a translation adjustment mechanism for aligning the cartridge stage and the assay cartridge relative to a vertical support structure, a fluid metering device, and a detection device for high throughput lateral flow assay analysis.

Description

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS

[0002] This application claims priority to U.S. Provisional Patent Application No. US62 / 784,065 filed on December 21, 2018, the entire contents of which are incorporated herein by reference. Technical Field

[0003] The present invention relates to the field of analytical chemistry, and more particularly to a lateral flow assay device analyzer that supports simultaneous diagnostic testing at multiple lateral flow assay devices. The present invention provides an integrated sample processing system for high throughput preparation and analysis of lateral flow assay samples. Background Art

[0004] Healthcare diagnostic laboratories use diagnostic instruments such as diagnostic analyzers to test and analyze samples. Healthcare professionals also use diagnostic tools at the point of care in clinics and other patient settings. One or more sample tubes or test devices are typically placed in a rack or carrier and loaded into the analyzer by a technician or operator. Known diagnostic analyzers use a variety of liquids and reagents to perform diagnostic analytical procedures. Analytical assays can be used for diagnostic applications, for example, in human health (e.g., blood and urine testing), environmental contamination (e.g., water and soil testing), and industrial food and drug preparations (e.g., bacterial contamination testing), but typically require extensive and expensive laboratory instrumentation and trained operators.

[0005] Lateral flow test strips based on the immunochromatographic principle exist for a variety of target analytes. The first lateral flow test was performed to detect human chorionic gonadotropin, and there are now commercially available immunological tests for monitoring ovulation, detecting infectious organisms, analyzing drugs of abuse, and measuring other analytes important to human physiology. Lateral flow assay products have also been introduced for veterinary testing, agricultural applications, environmental testing, and product quality assessment. While the first lateral flow assay tests gave qualitative results based on the presence or absence of a signal line, test design has evolved toward semiquantitative and quantitative assays with the integration of handheld readers and high-throughput analyzers. Most lateral flow test strips are modeled after existing immunoassay formats, typically sandwich assays, where the target antigen or target compound is immobilized between two layers of antibodies, a capture antibody and a detection antibody. In serum assays, antibodies are detected as indicators of various disease states and immune status, and the formation of a complex between free detector particles in the sample stream and the capture reagent bound to the membrane at the test line is detected. Other microfluidic paper-based analytical devices, called “μPADs,” can perform more complex tests with narrower flow channel dimensions in multiple flow directions (i.e., two and three dimensions, or 2D and 3D), as well as multiplex tests in parallel and, by extension, with smaller sample volumes than conventional paper strip tests of the past. The ability to work with smaller volumes is important when testing samples that are difficult to collect in large quantities, such as for point-of-care testing for human health.

[0006] Attempts have been made to reduce sample turnaround time and process a large number of lateral flow assay devices at one time to provide multiple testing and increase the throughput of assay testing and analysis. In one example, U.S. Pat. No. 9,389,228 issued to Jakubowicz et al. describes a metering mechanism for dispensing samples onto lateral flow assay devices, a radial incubator assembly having a plurality of receiving stations sized to individually receive a corresponding plurality of lateral flow assay devices, and a detection device for detecting test results from the lateral flow assay devices. In another example, U.S. Pat. No. 8,883,509 issued to Lemme et al. describes an apparatus for sequentially accessing and simultaneously processing a plurality of individually substrate-supported biological samples, the apparatus having substrate supports arranged in a relatively small arc, wherein individual substrate support units are automatically and independently movable between separate processing positions and separate access positions.

[0007] There remains a need for robust high throughput analyzers for lateral flow assay testing and analysis.

[0008] This background information is provided for the purpose of making known information believed by the applicant to be potentially relevant to the present invention. It is not necessarily intended, and should not be construed, as an admission that any of the foregoing information constitutes prior art with respect to the present invention. Summary of the invention

[0009] It is an object of the present invention to provide a high throughput analyzer for lateral flow assay testing and analysis.Another object of the present invention is to provide a combination cartridge system, analyzer and method for high throughput lateral flow analysis.

[0010] In one aspect, an assay device analyzer is provided, comprising: a box stage for releasably supporting an assay box containing a lateral flow assay substrate; a fluid metering device; a detection device; at least one vertical support structure for vertically supporting a plurality of assay boxes; a height adjustment mechanism coupled to the box stage to vertically align the height of the box stage; and a translation adjustment mechanism coupled to the box stage to translationally align the box stage relative to the vertical support structure and the fluid metering device.

[0011] In one embodiment, the height adjustment mechanism and the translation adjustment mechanism are independent.

[0012] In another embodiment, the fluid metering device is positionally adjustable to accommodate various configurations of the assay cartridge.

[0013] In another embodiment, the analyzer further comprises a second vertical support structure for vertically supporting a plurality of assay cartridges.

[0014] In another embodiment, the cartridge stage includes a stand for releasably grasping one or more components on the assay cartridge.

[0015] In another embodiment, the analyzer comprises a detection device.

[0016] In another embodiment, the analyzer comprises more than one detection device.

[0017] In another embodiment, the detection device is a fluorometer, a spectrophotometer, a colorimeter, a camera, a photomultiplier tube (PMT), a charge coupled device (CCD) camera, a digital camera using a complementary metal oxide semiconductor (CMOS) detector, a laser, or a photodiode.

[0018] In another embodiment, the fluid metering device is a sample metering device, a running fluid metering device, or a wash fluid metering device.

[0019] In another embodiment, the analyzer further comprises one or more additional fluid metering devices.

[0020] In another embodiment, the analyzer is enclosed in a housing.

[0021] In another embodiment, the housing comprises more than one assay device analyzer.

[0022] In another embodiment, the analyzer further comprises one or more of a temperature control device and a humidity control device.

[0023] In another embodiment, the vertical support structure is a box.

[0024] In another embodiment, the analyzer is a point-of-care analyzer, an automated clinical analyzer, or a combination thereof.

[0025] In another embodiment, the analyzer further comprises an electronic control system.

[0026] In another embodiment, the analyzer further comprises a quality inspection optical unit.

[0027] In another aspect, a method for flow assay analysis is provided, the method comprising: aligning a cartridge stage below a vertical support structure that holds a plurality of vertically disposed assay cartridges, each assay cartridge comprising an assay substrate having a detection area; raising the cartridge stage to engage the cartridge stage with the assay cartridge held in the vertical support structure; moving the cartridge stage with the engaged assay cartridge away from the vertical support structure and aligning it with a fluid metering device; metering fluid from the fluid metering device onto the assay substrate in the assay cartridge; incubating the assay cartridge; and detecting components of the sample at the detection area of ​​the assay substrate.

[0028] In one embodiment, the method further comprises dispensing one or more additional samples, buffers, reagents or detection components onto the assay substrate prior to incubating the assay cartridge.

[0029] In another embodiment, the method further comprises controlling the temperature and humidity surrounding the analyzer.

[0030] In another embodiment, the method further comprises adjusting the position of the fluid metering device and the detection device.

[0031] In another embodiment, the assay cartridge is moved by the cartridge stage to a second vertical support structure for incubation.

[0032] On the other hand, an assay device analyzer is provided, comprising: a box stage for supporting an assay box, the assay device comprising a planar support having at least one sample addition area, at least one reaction area, at least one detection area, and a wicking area disposed on the support, the wicking areas being fluidly interconnected along at least one lateral fluid flow path; a sample metering device; a detection device; a loading box for storing a plurality of assay boxes; a height adjustment mechanism coupled to the box stage for adjusting the height of the box stage relative to the loading box; and a translation adjustment mechanism coupled to the box stage for aligning the box stage relative to the loading box, the sample metering device, and the detection device.

[0033] In another aspect, a method for flow assay is provided, the method comprising: aligning a box carrier below a loading box at a height, the loading box comprising a plurality of analytical devices; engaging the box carrier with an assay device, the assay device comprising a planar support having at least one sample addition area, at least one reaction area, at least one detection area, and wicking areas interconnected along at least one lateral fluid flow path disposed on the support; sliding the box carrier away from the box and aligning it with a sample metering device; metering sample fluid from the sample metering device into the assay device; culturing the assay device; and detecting components of the sample on the detection device. BRIEF DESCRIPTION OF THE DRAWINGS

[0034] For a better understanding of the present invention and other aspects and features thereof, please refer to the following description used in conjunction with the accompanying drawings, in which:

[0035] FIG1A is a perspective view of a prior art flow assay cartridge device;

[0036] FIG. 1B is a diagram of various prior art flow assay micropad-type devices;

[0037] Figure 2 is a front perspective view of a flow assay analyzer;

[0038] Figure 3 is a close-up perspective view of a flow assay analyzer showing the dispensing and detection devices;

[0039] Figure 4 is a front perspective view of the analyzer translation adjustment mechanism, height adjustment mechanism and cartridge stage;

[0040] Figure 5 is a perspective view of a closed assay cartridge;

[0041] Figure 6 is a perspective view of the bottom of the assay cartridge;

[0042] Figure 7is a perspective view of an open assay cartridge;

[0043] Figure 8 is a view of the alignment of the height adjustment mechanism and the elevated cassette stage beneath the loading bin;

[0044] Fig. 9 is a view of a flow assay analyzer having a cartridge stage that carries a cartridge and translates beneath a dispensing and detection device;

[0045] Fig.10 is a view of the analyzer with the elevated cartridge stage aligned below the dispensing and detection device;

[0046] Fig.11 is a view of the analyzer with the cartridge stage lowered;

[0047] Fig.12 is a view of the analyzer with the cassette and stage translated into the incubator;

[0048] Fig.13 is a view of the analyzer with the stage elevated relative to the incubator; and

[0049] Fig.14 is a flow chart of a method of using the analyzer. DETAILED DESCRIPTION

[0050] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.

[0051] As used in the specification and claims, the singular form "a," "an," and "the" include plural references unless the context clearly dictates otherwise.

[0052] As used herein, the term “comprising” will be understood to mean that the following list is not exhaustive and may or may not include any other additional suitable items, such as one or more further features, components and / or elements, as appropriate.

[0053] As used herein, the term "sample" refers to a volume of a liquid, fluid, solution or suspension for the purpose of qualitatively or quantitatively determining any property or component thereof, such as the presence or absence of a component, the concentration of a component, and the like. As described herein, in the context of the present invention, a typical sample is derived from a human or animal body fluid, such as, but not limited to, blood, plasma, serum, lymph, urine, saliva, semen, amniotic fluid, gastric juice, mucus, sputum, mucus, tears, feces, and the like. Other types of samples are derived from human or animal tissue samples, wherein the tissue sample has been processed into a liquid, solution or suspension to display a specific tissue component for examination. Other non-limiting examples of samples that can be used are environmental samples, food industry samples, and agricultural samples.

[0054] As used herein, the terms "test device" and "flow assay device" refer to any device that receives a sample fluid and includes a fluid transport member or flow path, along which various areas or locations are provided for supporting one or more reagents, filters, etc., and the sample passes through the fluid transport member or flow path under the action of capillary action or other forces. The fluid path can be placed horizontally, vertically, or at any desired angle to obtain the best effect. Test devices include, but are not limited to, thin film or "dry slide" test elements, lateral flow assay devices, microfluidic paper-based analytical devices (μPAD), vertical flow assay devices, and chromatographic devices, collectively referred to herein as "flow assay devices."

[0055] As used herein, the term "analyzer" refers to any device capable of processing a variety of analytical tests or flow assay devices, and wherein multiple test devices can be processed. The analyzer also includes multiple components that are configured to load, culture, test, and evaluate multiple analytical test elements in an automated or semi-automated manner, and wherein the test elements are automatically dispensed and processed without user intervention. Analyzers include, but are not limited to, clinical diagnostic equipment and point-of-care testing devices.

[0056] As used herein, the term "reaction" refers to any interaction between a component of a sample and at least one or more reagents on or in a substrate of a test device or added to a substrate of a test device, or between two or more components present in a sample. The term "reaction" is used to define a reaction that occurs between an analyte and a reagent on a test device as part of a qualitative or quantitative determination of the analyte.

[0057] The term "substrate" or "support" refers to a support or matrix to which a sample is added and on or in which an assay is performed, or where a reaction between an analyte and a reagent occurs.

[0058] Described herein is a lateral flow assay analyzer for preparing and analyzing flow assay samples. The analyzer supports concurrent diagnostic testing on multiple flow assay devices and provides an integrated and powerful sample handling system with concurrent testing capabilities. The analyzer is designed to support the use of multiple flow assay devices so that multiple tests can be cultured and processed simultaneously. A sample metering device is provided to apply a metered amount of sample to each of the multiple test devices. Each flow assay device can also have different binding reactants to test different components in the sample fluid and can be labeled to identify its content. The present design provides a significant reduction in the size of the analyzer and reduces the overall footprint while providing a system for testing multiple components in a sample.

[0059] One advantage of the present system is that the flow assay analyzer described herein does not require the addition of wet reagents separately. In particular, dried reagents may have been incorporated into the structure of each flow assay device so that the sample fluid may be optionally added together or sequentially with another flow fluid or buffer, thereby allowing automated multiple test processing. The room temperature storage of the lateral flow assay device can also extend the shelf life. In addition, the lateral flow device and μPAD require only a very small amount of sample and other fluids and can be used with the present analyzer. In addition, each flow assay device processed by the analyzer can be prepared with different reagents and conjugates to detect different components of the sample, so that multiple sample components can be tested simultaneously. The application of the flow assay device described herein may also include the direct use of whole blood, thereby reducing the total processing time without centrifugation. In other uses, the sample can be diluted with an appropriate buffer, and an efficient detection method can be used for the diluted sample. By using the flow assay device in the analyzer described herein, the total reaction time that can be up to one hour using a standard non-automated device can be effectively reduced, preferably reduced to a range of five to ten minutes. The currently described automated flow assay device analyzer can be used in clinical as well as point-of-care analyzer applications and can reduce the overall cost of instrumentation and point-of-care sample processing. By further enabling sample multiplexing on a single flow assay device, assays for multiple reactants can be run simultaneously on a single flow assay device, thereby reducing the cost per test and significantly increasing effective throughput. Multiplexing can significantly increase throughput by an effective multiplexing factor and can be as much as 10-fold (or more).

[0060] The described flow assay device analyzer enables various internal controls and calibrations to be incorporated, thereby providing a means of ensuring calibration, quality of results, reproducibility, and the ability to track assay degradation over time. These features further provide a means of combining with other intelligent reliability systems such as those provided on smart clinical analyzers. Another advantage is that the stability of factory calibration or wet calibration allows factory calibration to simplify user operation. The analyzer currently introduced can also increase the calibration interval to a typical general chemical interval and provide commonality of formats between point-of-care testing (POC) and mainframe assays, thereby improving development. In this way, one assay can be used in two types of applications (POC and mainframe), providing higher throughput and economies of scale. Therefore, the above provisions ensure quality results and equivalent performance for the POC and mainframe markets. Thin film elements and flow assay cartridge devices can be used on the analyzer, with only a small adjustment to the analyzer stand, vertical support structure, and metering and detector positions, where the form factor of the flow assay device allows it to be interchangeable with thin film dry sliding elements and a large number of existing flow assay devices. The versatility is significantly enhanced, wherein systems can be realized that can combine thin film analytical test elements, lateral flow assay devices as described herein, and conventional wet chemistry systems or parts thereof into a single unit.

[0061] In the case where each box is unique relative to the reactants of the combination but the samples in the sample metering device are the same, the analyzer is simplified and each box can be marked, optionally with a bar code, which can be detected by an identification detection unit, optionally an optical unit. In this case, each box has a different binding conjugate, a single sample (e.g., blood) can be added to the sample addition area, each box measures different components in the sample, and optical tracking can be performed when the box passes through the analyzer from drying to running (cultivation) and then to analysis. In another embodiment, the sample can be preloaded on each box, and a metering mechanism can apply a certain amount of liquid (e.g., buffer or other running fluid) to each box to elute the sample. The identification device can be further installed to the analyzer, such as a barcode reader.

[0062] Various flow assay devices with various assay substrates can be used with the currently described automatic flow assay analyzer in clinical, analytical and / or point-of-care environments. To this end, it is apparent to those skilled in the art that the inventive concepts described herein are equally applicable to numerous other flow assay device designs and can be used for various other types of automated and point-of-care diagnostic clinical analyzers. Further, the automatic analyzer described herein can be configured to, for example, handle various flow assay devices without the need to include dry slide analysis elements as separate components, or alternatively include other analysis systems other than those for handling directional flow assay devices, as described herein, such as in conventional wet chemistry or chromatography systems. The currently described analyzer can be suitable for accommodating various cassette high-throughput devices that use liquid-attached lateral flow processing and detection, such as chromatography or slides, vertical flow assay devices, paper-based microfluidic analysis devices (μPADs), blot analysis, microfluidic boxes, and conventional immunoassays, chemical analyses, and thin film analysis test elements can also be tested. There is no particular limitation on the shape and size of the assay device, which may be any shape and size suitable for its intended use, and the analyzer may be adapted to accommodate assay devices of any size or shape. Multiple lateral flow assay devices may be used in conjunction with a lateral flow assay analyzer. A set of lateral flow assay devices may be interchangeable in an analyzer, and a set comprising multiple lateral flow assay devices may be easily loaded into an analyzer for parallel processing by the analyzer.

[0063] The design of the assay using the lateral flow device can be very different based on matrix or substrate materials and construction, liquid components and order of addition and assay time. Those skilled in the art will appreciate that various types of materials can be used for lateral flow assays, including but not limited to visualization materials, detection elements, labels or labeled binding materials, conjugate materials, detergents, blocking agents, analytes, enzymes, binding partners, capture elements, control substances and other detectable materials. Lateral flow devices are also most commonly used in immunoassays, although not necessary, and generally include one or more antibodies and antigens. Each substance in the assay can be suspended in a liquid to be added to a solid matrix, or applied to a solid matrix, so that the dissolution or suspension carried out by the added liquid promotes the movement of the substance along the matrix and combines with other substances. A variety of assay designs are known, which change the position and formulation of each assay material. On a solid matrix, the material can be arranged in different positions along the flow path, and arranged in the form of various points or lines, can be placed alone or mixed with other components, optionally can also be premixed in a liquid, then added to the matrix and the matrix is ​​dried before use. The substance can also be formulated in a mobile buffer or other liquid formulation at an appropriate time for addition to the solid matrix, depending on the assay design. The type and timing of liquid or reagent addition and the relative position of the solid components on the matrix can also greatly affect the results and sensitivity of the assay, and various configurations and combinations in conjunction with the assay design are known in the art and available to the skilled artisan as options for assay optimization.

[0064] The following exemplary embodiments relate to the configuration and design of lateral flow assay devices in the automatic lateral flow assay analyzer described herein. FIG. 1A shows an exemplary lateral flow assay device 100. The lateral flow assay device 100 according to this embodiment is defined by a planar substrate 102, which is preferably made of a suitable porous or non-porous material having surface properties that support capillary flow. In some embodiments, the planar substrate 102 may include a flow channel, optionally cutting into the surface of the substrate. The sample addition region 104 on one side of the lateral flow assay device 100 extends to at least one detection region 110, and preferably extends to a wicking region 106. The sample addition region 104 disposed at one end of the lateral flow assay device 100 forms a portion of a fluid flow path extending through a detection region 110 containing a detection conjugate or other reagent to detect a reaction with one or more components in the sample. The wicking region 106 at the opposite end of the fluid flow path attracts the sample fluid in the desired direction along the flow assay device 100. Optionally, the fluid flow path can also include additional separate areas containing reagents or detection conjugates, and other areas or positions that can be used for washing samples and any combined or uncombined components thereof along the path. Samples can be added to the test device by sample port 108, and results or reactions can be detected by result port 114. Extending from sample addition area 104 to wicking area 106 to create a defined fluid flow path, the area is optionally at least partially open. In another embodiment, the flow path is completely open. "Open" refers to the position above the planar assay substrate without a cover or covering. Therefore, if there is a cover as a physical protection for the flow path, a cover is not required to promote capillary flow in the flow path. In another embodiment, the planar substrate 102 is almost completely encapsulated by box 112 or housing, so that samples can be added at 104 sample addition areas and detection reactions at 110 detection areas through the holes in the box cover. Box 112 can accommodate and protect the flow assay device 100, and provide robustness for the delivery and movement of the flow assay device 100 in the analyzer. Solid box 112 can also be moved by the mobile mechanism of analyzer without damaging planar substrate 102 and detection zone therein.Box 112 can include box bottom, box sidewall and box end wall, to provide extra firmness and durability for lateral flow assay device.Can select to include covering or lid.Result port 114 in box is positioned around detection zone 110, so that one or more detectors can detect the reaction in detection zone 110.Various configurations of lateral flow assay device are known, including but not limited to device size, material, porosity of substrate, whether there is topographical feature on substrate, shape of passage and configuration and the variation of manufacturing method of passage.Throughout the specification, reference is made to a particular lateral flow assay device 100 according to an exemplary embodiment, however, it is apparent that other device designs and possible variations of such designs may also be similarly configured for interrelationship in a clinical analyzer as discussed herein.

[0065] The lateral flow assay device 100 is defined by a planar substrate 102, which includes a sample addition area 104 for receiving a sample from a liquid dispenser or a sample metering device. When the lateral flow assay device 100 is encapsulated by a box 112 or a box housing, the sample from the liquid dispenser is placed on the sample addition area 104 through a sample port 108. The sample is usually stored on the top of the sample addition area 104, and the sample addition area 104 is capable of transporting the liquid sample from the sample storage point to the reagent area through a conjugated pad 122. Once the fluid is added to the sample addition area 104 or the upstream of the sample addition area 104, the planar substrate promotes spontaneous lateral capillary flow along the defined fluid flow path between the sample addition area 104 and the wicking area 106. The lateral flow assay device may also include an optional filter material (not shown) that may be placed in the sample addition area 104 to filter particles from the sample or filter blood cells from the blood so that plasma can pass through the device. After the sample is delivered to the sample addition area 104, it will encounter the conjugate pad 122. After the sample flows through the conjugate pad 122 and interacts with the conjugate pad 122 and optionally with the reagent addition area, the sample and reagent plume will be included in the fluid flow. The reagent plume may include any reagent material dissolved in the conjugate pad 122 or any reagent material added by the optional reagent addition area. The reagent plume may include a conjugate having both a detection element and a binding partner, in which case it is generally referred to as a conjugate plume. Downstream from the detection area 110 along the fluid path is a wicking region 106 that is fluidly connected to the detection area. The wicking region 106 is an area of ​​the assay device 100 that has the ability to receive a liquid sample and any other material (e.g., unbound reagents, washing solutions, etc.) in the flow path. The wicking region 106 provides capillary forces to move the liquid sample through and out of the detection area of ​​the assay device. The wicking region may include a porous material, such as nitrocellulose. The wicking region may further include a non-capillary fluid drive device, such as using evaporative heating. Optionally, a hydrophilic foil or layer may be placed directly over at least a portion of the wicking region 106 to increase the overall flow rate or processing time of a sample applied to the flow assay device.

[0066] Components of the flow assay devices described herein, such as the physical structure of the device, whether or not separated from the rest of the device, can be made of, for example, copolymers, blends, laminates, metallized foils, metallized films, or metals, waxes, adhesives, or other suitable materials known to the skilled person. Alternatively, the device components can be made of copolymers, blends, laminates, metallized foils, metallized films, or metals deposited on any one or combination of the following materials: paraffin wax, polyolefin, polyester, styrene-containing polymer, polycarbonate, acrylic polymer, chlorine-containing polymer, acetal homopolymer and copolymer, cellulose and its ester, nitrocellulose, fluoropolymer, polyamide, polyimide, polymethyl methacrylate, sulfur-containing polymer, polyurethane, silicon-containing polymer, other polymers, glass, and ceramic materials. Alternatively, the components of the flow assay device can be made of plastic, elastomer, latex, silicon wafer, or metal. In one embodiment, the elastomer may include polyethylene, polypropylene, polystyrene, polyacrylate, silicon elastomer, or latex. Alternatively, the components of the device can be made of latex, polystyrene latex, or hydrophobic polymer. In one embodiment, the hydrophobic polymer may include polypropylene, polyethylene, or polyester. Alternatively, the components of the device may include Polystyrene, polyacrylate or polycarbonate. Alternatively, device components can be made of plastics that can be embossed, milled or injection molded, or of surfaces of copper, silver and gold films, on which various long-chain alkanethiols can be adsorbed. Plastic structures that can be milled or injection molded, such as cartridge housings, can include, for example, polystyrene, polycarbonate, polyacrylate or cycloolefin polymers.

[0067] The limited flow path of the assay device (including device 100) described herein may include an open or closed path, channel, groove and capillary for guiding the fluid to flow along the fluid flow path. The conjugate pad 122 may be optionally placed between the sample addition area 104 and the detection area 110 to fluidically connect these elements. The optional reagent addition area between the sample addition area 104 and the detection area 110 may also allow the addition of reagents from the outside of the device. For example, the reagent addition area may be used to add interruption reagents that can be used to wash other unbound components present in the sample and fluid flow path to the wicking area 106. Reagents may be added to the reagent area and may be dried on the reagent area before use, added to the reagent area using the reagent metering device on the analyzer before use, or both. Reagents may also be added by an optional reagent metering device. Reagents include but are not limited to binding partners, such as antibodies or antigens for immunoassays, DNA and RNA aptamers with or without resonance energy transfer (RET) and corresponding target analytes, substrates for enzyme assays, probes and auxiliary materials for molecular diagnostic assays, such as materials that stabilize integrated reagents, materials that inhibit interference reactions, etc. Typically, a reagent useful in the reaction carries a detectable signal as described herein. In some cases, the reagent can react with the analyte directly or through a series of reactions to form a detectable signal, such as a colored or fluorescent molecule. In a preferred embodiment, the reagent zone includes a conjugate material. The term "conjugate" refers to any portion with a detection element and a binding partner. In use, a fluid sample is introduced into the sample addition zone 104 in the device and will flow to one or more detection or test zones 110 within the fluid flow zone. The detection or test zone includes reagents useful for reacting with one or more target components in the sample zone or detecting target components in the sample zone.

[0068] For the purpose of description, a detection element is a reagent that is detectable with respect to its physical distribution and / or the intensity of the signal it transmits. Examples of suitable detection elements for use with the currently described assay devices include, but are not limited to, luminescent molecules, such as fluorescent agents, phosphorescent agents, chemiluminescent agents, bioluminescent agents, etc., colored molecules, molecules that produce color upon reaction, enzymes, radioisotopes, and ligands with specific binding, etc. Detection elements, also referred to as labels, are preferably selected from chromophores, fluorophores, radioactive labels, and enzymes. Suitable labels are available from commercial suppliers, providing a variety of dyes for labeling antibodies, proteins, and nucleic acids. For example, fluorophores span virtually the entire visible and infrared spectrum. Suitable fluorescent or phosphorescent labels include, for example, but are not limited to, fluorescein, Cy3, Cy5, and the like. Suitable chemiluminescent labels include, but are not limited to, luminal, transparent, and the like. Similarly, radioactive labels are commercially available, and detection elements can be synthesized so that they are incorporated into radioactive labels. Suitable radioactive labels include, but are not limited to, radioactive iodine and phosphorus; for example125 I and 32 P. Suitable enzyme labels include, but are not limited to, horseradish peroxidase, β-galactosidase, luciferase, alkaline phosphatase, and the like. Some other examples of detectable materials include, but are not limited to, colloidal gold, enzyme conjugates, other colloidal metals, fluorescent particles, and magnetic particles. For example, when multiple analytes or markers are detected, two or more labels can be used. A binding partner is a material that can form a complex that can be used to determine the presence or quantity of an analyte. For example, in a "sandwich" assay, a binding partner in a conjugate can form a complex comprising an analyte and a conjugate, and the complex can further bind to another binding partner, also referred to as a capture element, incorporated into the detection region. In a competitive immunoassay, the analyte will interfere with the binding of the binding partner in the conjugate to another binding partner (also referred to as a capture element) incorporated into the detection region. Exemplary binding partners included in the conjugate include, for example, antibodies, antigens, analytes or analyte mimetics, and other proteins.

[0069] The detection area 110 is a position where any detectable signal can be read on the test line 118 and the control line 120. The width in the flow path in the detection area 110 is generally on the order of 0.044-4mm, preferably on the order of about 2mm, although other widths of about 1mm can be prepared, provided that enough signals can be read to fit a suitable detection instrument. As described above, the capture element in the detection area 110 can retain the binding partner of the conjugate or the complex containing the conjugate. For example, if the analyte is a specific protein, the conjugate can be an antibody that will specifically bind the protein to the detection element (e.g., a fluorescent probe). The capture element can then be another antibody that also specifically binds to the protein. In another embodiment, if the label or analyte is DNA, the capture molecule can be, but is not limited to, a synthetic oligonucleotide, an analog thereof, or a specific antibody. Other suitable capture elements include antibodies, antibody fragments, aptamers, and nucleic acid sequences that are specific to the analyte to be detected. A non-limiting example of a suitable capture element is a molecule with an avidin functional group that can be combined with a conjugate containing a biotin functional group. The detection zone may also include multiple detection zones and include one or more markers.

[0070] FIG. 1B is a view of various prior art micropaper-based lateral flow assays, also known as micropad (μPAD) type devices, which are lateral flow assay devices that can be used with the present analyzer. The μPAD assay device has a sample addition zone or zone, and may have one or more reaction zones for detecting one or more components in the sample. μPAD substrates are widely used in analytical chemistry because they can provide an inexpensive and user-friendly analytical process suitable for miniaturization and small sample amounts. In the case of multiple detection zones, the capture element may include multiple capture elements, such as a first capture element and a second capture element. The conjugate may be pre-deposited on the assay device, such as by coating in the reagent zone. Similarly, the capture element may be pre-deposited on the assay device on the detection zone. Preferably, both the detection element and the capture element are pre-deposited on the assay device, or are deposited in the reaction zone and the detection zone, respectively. Capture elements, such as antibodies in the detection zone (e.g., by coating) and labeled conjugate materials that are also capable of participating in a reaction capable of determining the concentration of the analyte, are preferably stored on a device downstream of the sample addition zone 104, wherein the labeled conjugate material carries a label for detection in the detection zone.

[0071] Examples of applications of the currently described analyzer include, but are not limited to, biomedical diagnostics, such as pregnancy tests, glucose tests, biomarker tests, environmental tests, such as water tests for microorganisms or other contaminants (e.g., arsenic). Fluid samples that can be analyzed using the present device include, but are not limited to, water or aqueous samples from various sources (e.g., tap water, wells, pond / lake water, wastewater, rainwater, etc.), and body fluids such as blood, urine, saliva, sweat, tears, or amniotic fluid. The sample size used by the present analyzer may vary. In particular, the size of the analyzer and the device can be configured to accommodate sample sizes in the microliter range, such as samples less than 1000 μL and less than 10 μL, including samples less than 1 μL.

[0072] Antibodies used in immunochromatographic tests must have sufficient sensitivity, specificity, purity, and stability to accomplish the performance requirements of the final product. Depending on the assay design, the antibody can be used as a capture reagent on the test line, as a conjugate on the detector particle, or both. Purification and consistency of the supply source are also important. Since the antibody can bind to both the membrane and the detector particle, the proteins will compete for binding sites. There is also a decision as to whether to use polyclonal or monoclonal antibodies. Antibody preparations should be affinity purified at a minimum. Any ligand recognition system in which the detector particle is bridged to the capture reagent on the membrane can be used.

[0073] Figure 2It is a front perspective view of a flow determination analyzer 200, which includes a translatable box stage 202, a fluid metering device 204 and a detection device 206. A translation adjustment mechanism 216 is functionally connected to the box stage 202 so that the box stage moves along the x-x' axis in the analyzer component plane. It can also be imagined that other translation adjustment mechanisms and related mechanical components can be integrated to provide the box stage 202 along the translation of the y-axis normal to the x-x' and z-z' axes shown. The translation adjustment mechanism 216 can include a hydraulic actuator, an electric motor combined with a worm drive gear system, a movable track and gear, an electric servo motor or any other translation mechanism known to the technician. The height adjustment mechanism and the translation adjustment mechanism can be an independent mechanism that independently controls height and translation. Alternatively, the height adjustment mechanism and the translation adjustment mechanism can be a single mechanism that can control the translation and elevation motion of the box stage.

[0074] One or more vertical support structures are configured to vertically support a plurality of assay boxes in an assay analyzer. The minimum requirements of the vertical support structure include the position of the box to be maintained so that the assay boxes can be taken out from the vertical support structure one at a time, and provide a function that ensures that only the desired shuttle motion of the box carrier is utilized to take out the desired number of boxes from the stack or the desired number of boxes are added to the stack. The vertical support structure can be, for example, a vertical support cover, such as a box, one or more plates that are combined with one or more guide rails to guide the vertical position of the box, and at least a substrate or base support structure for positioning the box stack, and a holding mechanism keeps the box stack fixed, while enabling and guiding the required box to be moved by the box carrier. Optionally, the analyzer can include a single vertical support structure, wherein the movement of the assay platform can allow the box to be added to the top of the box stack in the vertical support structure, so that once all the boxes in the stack have been circulated through the analyzer and one or more vertical support structures, the first box eventually returns to the bottom. The assay analyzer can further have one or more additional shuttle mechanisms or a box carrier with a related translation and elevation mechanism. One or more vertical support structures can be optionally removed or reversibly engaged from the assay analyzer so that the assay box can be more easily loaded and / or removed. The vertical support structure can also pre-install the assay box to facilitate the technician to handle, or it can be selected for disposable use. In the described embodiment, the vertical support structure will be referred to as a box, but it should be understood that in the current described embodiment, other configurations of the described vertical support structure can be substituted. The loading box 212 used as the vertical support structure stores multiple assay device boxes for the analyzer in a vertical orientation so that the box stage 202 can be aligned with the bottom or loading box of the vertical support structure, and the stage mount 220 can releasably grip a single flow assay device box and slide the box out of the loading box 212. The stage mount can include, for example, one or more guides, columns, clips, friction devices or any other features that are reversibly attached to the bottom of the assay box 210. In other embodiments, the box stage 202 can be suitable for accommodating more than one assay box 210.

[0075] Fluid metering device 204 may include a syringe with a pump, such as a hydraulic or electric motor or servomotor with a suitable metering gear or motion device or fluid metering pump, for metering a fluid sample of an aliquot to the assay substrate inside assay box 210. The sample is preferably metered to the sample area by a syringe, which is operated by a controlled mechanical actuation, without contact or with minimal contact to prevent atomization of the sample fluid. Fluid metering device 204 preferably has an attached metering tip, which can be aligned with the sample port of assay box 210 by adjusting the height of the box stage 202 using a height adjustment mechanism 218. The height adjustment mechanism and translation adjustment mechanism shown are independent, however, they can also be a single translation device, which can translate and adjust the height of the box stage. Fluid metering device 204 can optionally be equipped with a metering tip that is optionally disposable. Analyzer 200 can also optionally have one or more additional or auxiliary metering devices 208, for metering a second fluid into the assay device, or multiple metering devices are set as needed. The example that can be metered into the fluid and fluid component of mensuration substrate includes but is not limited to sample, dilution sample, buffer, reagent, bonding agent, dyestuff etc.The mensuration device analyzer can also comprise more than two liquid metering tips, each is connected to identical or different liquid reservoir or liquid supply source, and each reservoir or supply source comprises identical or different liquid component, to add in solid matrix or mensuration substrate.Each liquid metering tip can be configured to for example convey the mensuration solution that comprises one or more components, optionally further comprises one or more mensuration materials, and wherein each metering tip can be transported to the lateral flow assay device place with liquid component or composition at the required time.As previously discussed, various solid matrix mensuration designs and structures are possible, and can be accommodated by the flow assay analyzer of current description.

[0076] The electronic control system of the microcontroller included in or connected to the assay device analyzer can also be configured to control one or more electronic controllers, motors and pumps of the flow assay analyzer, including but not limited to the motion and position of the control components, the order of addition of the liquid components, the amount of liquid added, the time of moving or adding liquid, the moving speed, the liquid addition and the detection device control including the detection parameters. The control system can be further controlled or set using a local or remote user interface or control device, which can control the addition of fluid to the assay device according to the desired assay design, and can include a memory and an input device for receiving and storing data about the assay design and conditions. The control system can be configured to communicate with one or more user interfaces by being connected to the control system, and the user interface can include but not limited to a remote terminal, a desktop, a laptop, a tablet computer, a smart phone or an intelligent electronic device. The control system can be further connected to a communication network by a wired or wireless connection, and the wired or wireless connection is, for example, a connection device supporting wi-fi, a Bluetooth connection device or other devices supporting communication. The electronic control system can also optionally further communicate with a processor connected to the detection device by wire or wirelessly. Each in fluid metering device 204 and one or more auxiliary metering devices 208 can be further connected to electronic control system, to control one or more of the volume, time and position of adding fluid on assay matrix or assay device.Once sample is added in assay box 210 by fluid metering device 204, translation adjustment mechanism 216 is just in translational alignment box stage 202 below incubator 214 used as second vertical support structure, wherein can store assay box device in culture process.In the embodiment with single vertical support structure, translation adjustment mechanism and height adjustment mechanism can move assay box to put it back in single vertical support structure, preferably be placed on the top of vertical support structure, make it possible to unload next assay box from the bottom of vertical support structure.As used herein, term " cultivation " refers to the time between sample or flowing fluid being added to the time that can detectable component is combined with detection zone on planar substrate.Cultivation time can also be considered as the time required between adding sample and / or buffer and the time that can obtain assay device result.Cultivation time will be different according to the material, thickness and size of the size and property of planar substrate and the flow assay device used.

[0077] After cultivation, the detection device 206 attached to the detection instrument can detect the perceptible signal of the detection area of ​​the flow determination device and visualize the result of the flow determination. The detection device 206 is arranged and aligned to scan the test area of ​​the flow determination device positioned in the adjacent test area. Preferably, the detection device 206 is electrically connected to a processor for image processing. This analyzer can use other detection devices, and its examples include but are not limited to fluorometers, spectrophotometers, colorimeters, cameras, photomultiplier tubes (PMT), charge coupled devices (CCD) cameras, digital cameras using complementary metal oxide semiconductor (CMOS) detectors, lasers and photodiodes. In one embodiment, a camera equipped with a single LED or LED array is used to optically scan the linear portion of the sample addition area and the wicking area along the part of the fluid flow path and preferably along the flow channel, and preferably includes the detection area, and at least one reaction area includes a test line and preferably also includes a control line, depending on the construction of the element. These detection methods can be used in combination with any other optical method such as a lens, a filter or a signal-to-noise ratio for improving the signal or the detected signal.

[0078] The box stage 202 can hold at least one flow assay device. The box stage 202 grasps and holds the assay device via the feature in the stage mount 220, and aligns the flow assay device with the sample metering device, the detection device and the vertical support structure during cultivation and analysis. The stage mount 220 can also be designed to hold various box configurations, or interchangeable on the box stage 202, so that it can releasably engage various boxes. By releasably engaging with the stage mount 220, the lateral flow assay box is distributed from the lower opening of the loading box 212, and it is safely maintained within or on the stage mount 220 on the box stage 202, so that it can be transported in the assay analyzer. The stage mount 220 can engage the bottom and / or side edge or lateral edge of the assay box 210, so that it can be releasably grasped and moved translationally. Any physical configuration of the stage mount and the assay box can be used, wherein the stage mount can releasably and safely hold the assay box, including but not limited to features for snap-in, sliding or gripping engagement. The assay box is shuttled back and forth on the box stage 202 to the metering and detection area by a translation adjustment mechanism, wherein the assay box is preferably moved to the metering area 202 from the loading box 212 one at a time. The height adjustment mechanism 218 is functionally connected to the box stage 202, and moves the box stage up and down along the axis z-z', so that the box stage 202 and the stage mount 220 are aligned with the loading box 212, the incubator 214, so that the stage mount 220 can engage and separate each assay box, and align the assay box by one or more sample metering devices and one or more detection devices. The box 212 and 214 near the box stage can accommodate a plurality of stored lateral flow assay devices. If desired, a plurality of boxes and / or other vertical support structures can also be used. The position of the metering device relative to the stage, reservoir, detection device and other features and the relative position of the metering device and the detection device can be adjusted to adapt to different lateral flow determination device configurations and different distances between the sample addition area and the detection area, as well as the box or device size (such as the width and length of the device). The analyzer may include one or more metering devices to deliver more than one sample, reagent, buffer or other fluid to the device. The fluid delivered by the metering device may include any aqueous or non-aqueous fluid, optionally including sample, buffer, reagent and / or detection component. The flow determination instrument may be further contained in a housing providing temperature and / or humidity control, and the housing may further include one or more heaters, coolers and / or humidity control devices. The housing may also accommodate one or more flow determination instrument devices.

[0079] Figure 3is a close-up perspective view of the flow analyzer showing the dispensing and detection apparatus, with close-ups surrounding the metering and detection areas of the analyzer. The vertical support structure stores a plurality of vertically stacked assay cartridges, and each assay cartridge 210 can be individually removed one at a time by sliding through a side hole 222 of the vertical support structure after the bottom of the assay cartridge 210 passes through the side of the vertical support structure to engage with the stage mount 220 on the cartridge stage 202. The stage mount 220 has features that releasably interact with and retain the bottom of each assay cartridge 210 so that the assay cartridges can be slid out of the vertical support structure along the translation axis x-x' using a translation mechanism. As shown Figure 3 As shown, when the analyzer is used, the assay cartridge is engaged with the stage mount 220 and slid off the vertical support structure by the translation adjustment mechanism, and then the cartridge stage 202 is aligned along the x-x' axis translationally and vertically aligned along the axis z-z' using the height adjustment mechanism, so that the sample port of the assay cartridge is aligned with the fluid metering device 204, where the sample is applied to the assay cartridge 210 through the sample port. The optional auxiliary metering device 208 provides a fluid distribution system for dispensing auxiliary fluid to the assay cartridge as required by the individual assay being run, and can be optionally aligned with the auxiliary metering port on the assay cartridge. The detection device 206 is shown as being positioned near the fluid metering device 204, but can be in any position where the assay cartridge can be positioned and aligned for effective detection by the translation adjustment mechanism and the height adjustment mechanism.

[0080] Figure 4 2 is a front perspective view of an embodiment of an analyzer translation adjustment mechanism 216, a height adjustment mechanism 218 and a box stage 202. The height adjustment mechanism 218 has a motor or other mechanism to move the box stage 202 with the stage mount 220 up and down along the axis z-z' through a functional connection with the lifting rod 230. Optional alignment posts 232a and 232b help control the vertical alignment of the box stage 202 and prevent the box stage 202 from being misaligned with the x-x' axis. The housing of the height adjustment mechanism 218 can provide further guide means in the form of a slot, a hole or an elongated hole to guide the up and down movement and alignment of the alignment posts 232a and 232b and the lifting rod 230 to maintain the precise positioning of the box stage 202 in the analyzer.

[0081] Figure 5It is a perspective view of an exemplary closed assay box 250 used with the analyzer currently described. The assay box 250 includes a box body 260 with a bottom wall and a side wall, and the box body 260 can be loaded with a thin film analysis test element or a substrate. The assay box shown has an optionally removable box cover 258, which includes a sample port 252 for receiving a fluid sample, an auxiliary fluid port 254 (optional according to the requirements of the assay) and a result port 256, for providing an opening, and the analysis result can be detected by the detector through the opening. The assay box 250 can also have one or more optional bar codes 262, which can be any digital data stored as an image that can be read by an optical reader. Alternatively, the assay box can have one or more other identification tags, such as an RFID tag or an electromagnetic tag.

[0082] Figure 6 2 is a perspective view of the bottom of the assay box 250 with the box body 260. The features of the bottom of the assay box enable engagement with the stage mount for holding the assay box so that the box stage can align the assay box device with the sample metering device and the detection device. In the illustrated embodiment, the assay box has two flexible arms that engage the stage mount to allow the box to snap into place. These same flexible arms also allow the cartridge to slide in or out of the stage mount.

[0083] Figure 7 is a perspective view of an open assay cartridge 250 with the cartridge lid 258 open and angled relative to the cartridge body 260 and with an optional bar code 262. Also shown in the cartridge lid are a sample port 252, an optional auxiliary fluid port 254, and a result port 256.

[0084] An exemplary method involving testing at least one lateral flow assay device in an assay analyzer 200 is described herein. Figures 8 to 13 The movements of the analyzer are shown sequentially in this exemplary method as the assay cartridge is shuttled through the analyzer from storage to sample distribution to incubation to analysis. Figure 8 start, Figure 8 2 is a close-up view of the lifting mechanism aligning the box stage raised below the loading box, and the translation adjustment mechanism aligns the box stage below the loading box containing the assay boxes. After alignment, the height adjustment mechanism adjusts the height of the box stage by raising the lifting rod along the z-z' axis to align the stage mount on the box with the bottom of the bottom-most assay box in the loading box, as shown by the arrow. The bottom of the assay box reliably engages the features on the stage mount 220 to support the transportation of the assay box on the stage mount 220 throughout the movement of the assay box in the analyzer. The assay boxes are dispensed from the underside opening of the loading box and retained in the stage mount on the box stage.

[0085] Fig. 9is a close-up view of the analyzer, with the cartridge carrier holding the cartridge and translating beneath the dispensing and detection device. Fig. 9 As shown, once the stage mount is reliably engaged with the bottom side of the assay box, the box stage 202 is shuttled laterally along the x-x' axis so that the assay box slides out of the loading storage box through the box side hole 222 along the direction shown by the arrow. The assay box is reciprocated along the x-x' axis translationally, and the assay box is preferably moved from the loading box 212 to the box stage 202 one at a time. Adjacent to the loading box is the metering area of ​​the analyzer, which includes a fluid metering device 204 and optionally includes one or more auxiliary metering devices. The lateral flow assay device box on the box stage 202 is shuttled into the metering area by the translation adjustment mechanism 216, so that the assay box is located within the range of the sample addition area, and the sample port of the box is aligned with the fluid metering device 204. Then a certain volume portion of the sample contained in the metering tip is distributed on the dry slide test element to start flowing in the flow assay device.

[0086] Fig.10 It is a close-up of the analyzer, in which the box stage of elevation is aligned below the distribution and detection device. Once aligned under the metering area, the height adjustment to the box stage 202 provided by the height adjustment mechanism is utilized to move the assay box to the appropriate position and align with the sample metering device along the z-z' axis. The sample of the predetermined volume is then deposited on the sample addition area of ​​the lateral flow assay device. Based on the design of the lateral flow assay device, applying the sample to the sample addition area can spontaneously induce the distributed patient sample to produce lateral capillary flow along the limited flow path, or can start by adding other running fluids. In the assay device, then, under the effect of the capillary force produced along the flow path extending through the reaction zone on the substrate, the sample flows out from the sample addition area.

[0087] Fig.11 is a close-up of the analyzer with the cartridge stage lowered after sample addition. When the fluid sample first combines with the detection conjugate or other reagent, the sample begins to dissolve the conjugate, thereby generating a perceptible plume indicative of process flow, such as a conjugate plume. The height adjustment mechanism then lowers the cartridge stage 202 along the z-z' axis to below the height of the incubator to allow the translation adjustment mechanism to slide the cartridge stage 202 below the incubator.

[0088] Fig.12is a close-up of the analyzer with its box and stage translated along the x-x' axis to the incubator. During the incubation period, the sample and related materials advance toward the detection area and wicking area of ​​the assay device. The fluid sample continues to flow along the flow path and any intermediate reaction areas, thereby enabling a reaction to occur, which can be detected by the detection device. The sample continues to advance to the wicking area, which is sized to accommodate the amount of fluid dispensed. The flow assay device is then incubated for a predetermined time based on the flow assay device and the assay is performed.

[0089] Fig.13 is a close-up of the analyzer with the stage raised along the z-z' axis relative to the incubator to deposit the assay cartridges into the incubator until they are removed for reading. An orifice in the bottom of the incubator receives the assay cartridges from below and features in the lower portion of the incubator secure the cartridges. The cartridges are retained in the incubator for a predetermined residence time or incubation time depending on the number and type of tests to be performed on the assay device and the elution or detection time required for the assay. After the predetermined incubation time (e.g., 5 minutes, 10 minutes, etc.), the analyzer aligns the cartridge stage and stage mount over the lowest assay cartridge in the incubator and positions the cartridges in the incubator in the same manner as the cartridges in the incubator. Figure 8 and Fig. 9 As shown, the assay box is slidably removed from the incubator through the side opening in the incubator in a similar manner to the loading box. Once the assay box is adjusted by translation under the detection device, the result is read to allow the sample and reactant contained to be detected and scanned along the detection area of ​​the alignment assay device of the flow path. Optionally, a height adjustment mechanism can be used to align the assay box at the optimal position for detecting in the detection area on the assay box. Once the detection is completed, the box is taken out from the box stage to process other assay boxes. In one embodiment, the assay device can be slid off the box stage and dropped by the vertically arranged outlet chute (not shown) and discarded. The determination test instrument with a suitable detection device can determine the determination result, which can include the presence and / or concentration of analyte in the sample solution. Therefore, the analyte detection performed by the detection device that allows the analysis test or the detection result can be provided in real time. Further analysis (optionally using a prediction algorithm) can be processed in an onboard or connected computer.

[0090] Fig.14It is a flow chart of the method using an analyzer. First, the stage mount on the box stage is raised so that it is engaged with the bottom of the assay box stored in the loading box 302. Once engaged, the box stage with the combined assay box slides out of the side of the loading box and is aligned with the sample metering device 304. Then the sample is metered into the assay box 306, wherein the sample of known amount or volume is stored on the assay matrix or substrate element. Then the assay box is allowed to cultivate the determined time amount according to 308 assays. Once the cultivation is completed, the assay box is aligned with the detection device 310, and the result of the assay is determined with the detection device 312, wherein the test instrument determines the result (analyte concentration, detection, etc.). The cultivation of one or more assay boxes may occur, and other boxes can load samples simultaneously, to provide a simplified method for performing multiple assays in the same time period. In this embodiment, the box in the cultivation can be loaded into the second box, and once the sample has been added in all boxes, all boxes can be moved back to the first box, so that each box can be removed and the result can be read by the detector in the sample addition order.

[0091] The flow determination instrument currently described may also include a quality inspection (QC) optical device unit. The quality inspection optical device may be used for a variety of purposes in an automated flow determination instrument, including but not limited to identifying and / or verifying a unique analysis box, and confirming that the correct number and quality of samples and / or reagents have been added to the flow determination substrate. The optical unit for quality inspection may be used independently of the detection device or used together with the detection device to detect the measurement results and the process of the analysis device or its parts or the movement of the determination box through the analyzer. The quality inspection optical unit may include one or more optical detection devices for optical interrogation and detection of various aspects of the flow determination process, including one or more cameras, scanners and detection devices, for monitoring and quality inspection of the various steps performed by the high-throughput flow determination instrument. In some non-limiting embodiments, the optical unit may have a single photodetector or a group of photodetectors, for example, including one or more photoresistors, photodiodes, phototransistors, complementary metal oxide semiconductor (CMOS) image sensors and / or charge coupled device (CCD) image sensors. The optical unit may further include one or more light sources, for example, one or more light emitting diodes (LEDs), bulbs, mercury lamps and lasers. The data reader LED can also be used as an optical data reader to detect and read coded digital data on the assay box, such as bar code data, QR (quick response) code data or other identification marks to reliably identify the assay box housing. The QC optical device can also store and read data related to the operation of the assay box when the assay box is located in the analyzer, including but not limited to volume data of sample and reagent addition, buffer addition, addition and analysis time, and the collected data data can be related to other data, such as reagent and sample batch information, patient information, assay and substrate configuration and / or setup information, and other information related to the assay to provide additional quality data for each assay. The optical unit can also optionally be one or more passive optical components, such as one or more lens assemblies, reflectors, prisms, diffusers, optical fibers, waveguides, collimators, beam splitters, gratings, polarizers, and one or more optical filters, such as short pass, band pass, long pass, band stop, notch filter or other optical filters. The QC unit may also perform in-process checks on one or more objects or devices in the flow assay analyzer, including but not limited to the sample syringe, secondary syringe or reagent syringe, syringe cap, syringe plunger, sample quality or quantity in the syringe, position or status of the test cartridge, barcode on the cartridge, size and physical properties of sample or reagent dispenser sample droplets, and housing / fixtures housing the above components. In one embodiment, the optical unit may detect changes in the sample and / or reagent fluid by measuring one or more of fluid color, turbidity, reflectivity, volume, and droplet size or shape.The QC unit can also be used to determine the presence of a sample syringe, the presence and position of a syringe cap and / or plunger, the presence of a syringe needle holder, the presence, position and / or volume of bubbles and / or air gaps inside a sample or fluid metering device or any other additional sample or fluid metering device, the sample amount, the color and / or color uniformity of the sample, the presence and volume of a dispensed sample drop, the presence and volume of a dispensed reagent, the presence and position of a box, and the barcode information on the box. An optical unit located close to the fluid metering device can monitor, detect, record and possibly provide feedback on the condition of the fluid in the metering device in the analyzer. In an exemplary configuration, the QC optical unit consists of one or more light sources and one or more optical detectors located at the end of an optical path, and the object or device to be inspected is located between the light source and the optical detector along the optical path. The optical detector can detect photons from the light source, reflected and / or transmitted by the object and / or generated by the object. Depending on the nature of the light source and the optical detector, the optical path can be in different orders and / or combinations in free space or inside or on the surface of a passive optical component, reflected or deflected by a passive optical component. Various combinations of light sources and optical detectors can also be used. The optical unit may also monitor the fluid metering device to monitor fluid dispensing, for example, by monitoring the size and / or properties of the fluid or droplets and the general fluid flow inside the fluid metering device (e.g., air gaps and / or bubbles that may occur in a syringe-type metering device, which may produce false negatives in the assay if an insufficient amount of sample is metered onto the assay substrate). When the concentration and volume of the sample used are small to achieve high throughput, metering errors can lead to poor results and insufficient quality and reproducibility. Bubbles or air gaps in the reagent metering system or device and any operating fluid or buffer metering device may cause similar situations. Therefore, the quality control optical unit can monitor one or more delivered fluids to ensure fluid quality and correct quantity, thereby improving the reproducibility and confidence of the results.

[0092] All publications, patents and patent applications mentioned in this specification all indicate the technical level of those skilled in the art to which the present invention belongs, and are incorporated herein by reference. Having thus described the present invention, it is apparent that the present invention may be modified in many ways. Such modifications should not be considered as departing from the scope of the present invention, and all such modifications apparent to those skilled in the art are intended to be included within the scope of the appended claims.

Claims

1. Assay device analyzer, comprising: a cartridge stage for releasably engaging a lateral flow assay cartridge, the cartridge stage comprising a scaffold having features for releasably grasping one or more elements on a bottom of the lateral flow assay cartridge housing a lateral flow assay substrate; First and second vertical support structures for vertically supporting a plurality of vertically stacked lateral flow assay cartridges, the first and second vertical support structures comprising: a base support structure for supporting the plurality of vertically stacked lateral flow assay cartridges, including features for retaining the lateral flow assay cartridges and having an aperture in a bottom portion to allow engagement of a bottommost lateral flow assay cartridge in a vertical support structure by lifting the rack on the cartridge stage and secure engagement of the bottom portion of the lateral flow assay cartridge with the features on the rack; and a side aperture for enabling a bottom-most lateral flow assay cartridge to be slid into and out of the side aperture of the vertical support structure after the bottom-most lateral flow assay cartridge has been engaged with the carriage and the carriage has been translated away from the side aperture, each lateral flow assay cartridge being individually removable one at a time by sliding through the side aperture of the vertical support structure; and a holding mechanism configured to hold a plurality of vertically stacked lateral flow assay cartridges stationary while enabling a bottommost lateral flow assay cartridge to be moved out of the side aperture by the cartridge stage; a fluid metering device disposed between the first and second vertical support structures; a height adjustment mechanism coupled to the cassette stage to move the cassette stage up and down in a zz' axis to vertically align and elevate the cassette stage below the first and second vertical support structures; and A translation adjustment mechanism is coupled to the cartridge stage to translationally align the cartridge stage relative to the vertical support structure and the fluid metering device along a translation axis xx' perpendicular to axis zz'.

2. The analyzer according to claim 1, wherein: The fluid metering device can be positionally adjusted to accommodate various configurations of the assay cartridge.

3. The analyzer according to claim 1 or 2 further comprises a detection device.

4. An analyzer according to claim 3, comprising more than one detection device.

5. The analyzer according to claim 3, wherein: The detection device is a fluorometer, a spectrophotometer, a colorimeter, a camera, a photomultiplier tube (PMT), a charge coupled device (CCD) camera, a digital camera using a complementary metal oxide semiconductor (CMOS) detector, a laser, or a photodiode.

6. The analyzer according to claim 1 or 2, wherein: The fluid metering device is a sample metering device, a running fluid metering device or a washing fluid metering device.

7. The analyzer of claim 1 or 2, further comprising one or more additional fluid metering devices.

8. The analyzer according to claim 1 or 2, wherein: The analyzer is enclosed in a housing.

9. The analyzer according to claim 8, wherein: The housing includes one or more assay device analyzers.

10. The analyzer according to claim 1 or 2, further comprising one or more of a temperature control device and a humidity control device.

11. The analyzer according to claim 1 or 2, wherein: The vertical support structure is a box.

12. The analyzer according to claim 1 or 2, wherein: The analyzer is a point-of-care analyzer, an automated clinical analyzer, or a combination thereof.

13. The analyzer of claim 1 or 2, further comprising an electronic control system.

14. The analyzer according to claim 1 or 2, further comprising a quality inspection optical unit.

15. A method for flow assay analysis, the method comprising: aligning a cartridge stage beneath a vertical support structure having a bottom aperture, the vertical support structure holding a plurality of vertically disposed assay cartridges, each assay cartridge comprising an assay substrate having a detection area, the cartridge stage comprising a cradle for releasably grasping one or more components on the assay cartridges; raising the cartridge stage in the z-z' axis so that the cartridge stage engages the bottommost assay cartridge held in the vertical support structure through the bottom hole of the vertical support structure; The assay cartridge has a cartridge bottom for securely engaging a feature on the rack to support transport of the assay cartridge on the rack; moving the cartridge stage with the engaged assay cartridge along a translation axis x-x' perpendicular to the axis z-z' through the side hole in the vertical support structure and away from the vertical support structure and into alignment with the fluid metering device; metering the fluid from the fluid metering device onto the assay substrate in the assay cartridge; culturing the assay kit; as well as A detector is used to detect components of the sample at the detection zone of the assay substrate.

16. The method of claim 15, further comprising, prior to incubating the assay cartridge, dispensing one or more additional samples, buffers, reagents, or detection components onto the assay substrate.

17. The method of claim 15 or 16, further comprising controlling the temperature and humidity surrounding the analyzer.

18. The method of claim 15 or 16, further comprising adjusting the position of the fluid metering device and the detector.

19. The method according to claim 15 or 16, wherein: The assay cartridge is moved by the cartridge stage to a second vertical support structure for incubation.

Images