Microfluidic test strip chip and preparation and use thereof

Abstract

A microfluidic detection test paper chip, comprising a substrate, a microfluidic channel, and a dot array arranged reagent block, the microfluidic channel and the dot array arranged reagent block are arranged on the substrate, and are used for detecting enzymes, chemicals, proteins, polypeptides, amino acids, nucleic acids, and exosome components in a liquid sample; a syringe is used to inject a sample or a reagent into the microfluidic channel and infiltrate the reagent block; a color reaction is completed in the reagent block; color reaction data of the dot array arranged reagent block is scanned by an optical sensor to obtain a detection result, wherein the microfluidic channel and the reagent block are manufactured by using a microfabrication process, the reagent block is printed into a dot array arranged groove formed by the substrate and the microfluidic channel, and thus a micro-specimen multi-index analysis and detection effect is achieved.

Description

Technical Field

[0001] This application relates to the field of medical testing consumables, specifically to a microfluidic test paper chip and its preparation process and usage. Background Technology

[0002] Dry chemical test strips are widely used in medical testing. Integrated test strips made using printing or inkjet printing technologies can simultaneously detect hundreds of indicators on a single strip (Patent Publication No.: CN112362648A). Compared with conventional wetting or smearing methods, wetting test strips using spraying technology can significantly reduce the amount of sample used (Patent Publication No.: CN112505027A). However, spraying technology still cannot meet the needs of detecting more indicators with small amounts of sample.

[0003] In theory, microfluidic technology can precisely wet each reagent block of a test strip with trace amounts of sample or reagent. However, the complex manufacturing process of three-dimensional microfluidic channels and the heavy substrate of the test strip increase costs, which hinders the widespread adoption of microfluidic dry chemical test strips in medical testing.

[0004] Therefore, a planar microfluidic channel is constructed and bonded to the test strip substrate. By changing the valve size, the flow rate and direction of the sample or reagent entering the reagent block are controlled. The reagent block is positioned on the integrated test strip according to the target molecular weight of the detected index. This achieves a microfluidic technology solution that precisely wets the integrated test strip reagent block with trace amounts of sample or reagent, thus achieving the technical effect of detecting more indicators with low-cost microfluidic test strip chips. Summary of the Invention

[0005] (a) The technical problems to be solved.

[0006] This invention provides a microfluidic test paper chip, its manufacturing process, and application method. A planar microchannel structure is constructed and bonded to a substrate, and a reagent block is printed on the substrate to produce a low-cost microfluidic test paper chip. The flow rate and direction of the sample or reagent entering the reagent block are controlled by changing the valve size of the microchannel. The position of the reagent block on the substrate is precisely set according to the target molecular weight of the indicator detected by the reagent block, so as to achieve precise control of sample or reagent wetting of the integrated test paper strip reagent block.

[0007] (ii) Technical solution.

[0008] On one hand, one embodiment of this application provides a microfluidic test paper chip for multi-index detection of trace samples, including a substrate, microfluidic channels, and a reagent block. The microfluidic channels are bonded to the surface of the substrate and are used to control the flow rate and direction of liquid samples or reagents. The microfluidic channels include a sample dispensing component, at least one first port, at least one capillary network, and multiple second ports. The sample dispensing component is connected to the first port, the first port is connected to the capillary network, and the capillary network is connected to the second ports. The capillary network forms multiple grooves arranged in a dot matrix on the substrate, and each groove is connected to the capillary network through a second port. The reagent block is disposed in the grooves constructed by the substrate and the microfluidic channels. The reagent block includes a reaction component and a waste liquid absorption component for performing sample and / or reagent colorimetric reactions.

[0009] Furthermore, the sample dispensing assembly includes at least one sample orifice, the sample orifice including a first interface for connecting a syringe to dispense a liquid sample.

[0010] Furthermore, the sample dispensing assembly includes at least one sample orifice, the sample orifice including a first filter screen for filtering large particulate components in the liquid sample.

[0011] Preferably, the sample dispensing assembly includes two or more sample wells for dispensing different samples from the same individual, or the same type of samples from different individuals.

[0012] Furthermore, the sample application assembly includes an extension tube, the proximal end of which is connected to a first interface, and the distal end of which is pluggably connected to a first port.

[0013] Furthermore, the sample dispensing assembly includes at least one reagent port, which includes a second interface and a second extension tube for connecting a syringe and dispensing reagents. The proximal end of the second extension tube is connected to the second interface, and the distal end is pluggably connected to the first port.

[0014] Furthermore, the sample dispensing assembly includes at least one elastic reservoir for storing liquid samples or reagents and slowly and continuously injecting the liquid samples or reagents into the capillary network through the first port. The elastic reservoir includes an injection vessel, a vessel body, and a valve. The injection vessel is used to connect to a syringe needle, the injection vessel is connected to the vessel body, and the vessel body is pluggably connected to the first port through the valve.

[0015] Furthermore, the microfluidic pipeline includes multiple microvalves, which are disposed between the second port and the groove and are one-way valves used to control the flow direction of liquid samples or reagents.

[0016] Preferably, the second port includes a first-stage second port, a second-stage second port, a third-stage second port, and a final-stage second port. The first-stage second port has the smallest opening size and is used to connect the capillary network with the groove near the first port area. The final-stage second port has the largest opening size and is used to connect the capillary network with the groove away from the first port area.

[0017] Furthermore, the reagent block includes a filter membrane component disposed between the reaction component and the second port for filtering large particulate components or components that interfere with the colorimetric reaction in the detection sample.

[0018] Preferably, the reagent block is disposed in the groove formed by the substrate and the microfluidic channel, which includes determining the position of the reagent block in the groove on the substrate based on the target molecular weight of the index detected by the reagent block. The reagent block with a large target molecular weight of the index detected by the reagent block is disposed in the groove near the first port region, and the reagent block with a small target molecular weight of the index detected by the reagent block is disposed in the groove away from the first port region.

[0019] On the other hand, one embodiment of this application provides a method for preparing a microfluidic test paper chip, which includes the following steps.

[0020] Step 1: Design the microfluidic test paper chip, including designing the microfluidic pipeline circuit diagram, selecting the type and type of reagent blocks, and arranging the reagent blocks on the substrate.

[0021] Step 2: Microfabricate microfluidic channels and reagent blocks.

[0022] Step 3: The microfluidic channels are bonded to the substrate, and the capillary network forms a groove lattice on the substrate.

[0023] Step 4: The reagent block is printed onto the substrate groove matrix.

[0024] Step 5: Install the extension tube and elastic reservoir, and place them into the detection box.

[0025] On the other hand, one embodiment of this application provides a method for using a microfluidic test paper chip, which includes the following steps.

[0026] Step 1: Select the microfluidic test paper chip.

[0027] Step 2, add the sample.

[0028] Step 3, add reagents.

[0029] Step 4: Control the reaction conditions.

[0030] Step 5: Scan and detect, then obtain the results.

[0031] (iii) Beneficial effects.

[0032] (1) The combination of microfluidic technology and low-cost, easy-to-use integrated test strip technology enables liquid samples or reagents to be precisely "permeated" to saturate the integrated test strip reagent block, thereby replacing the existing "spraying" or "flooding" methods and achieving the effect of detecting hundreds of indicators with a small amount of sample.

[0033] (2) Compared with existing integrated test strip technology, the precise "permeation" method of microfluidic technology can ensure that each reagent block is fully permeated. Therefore, more reagent blocks of different types can be integrated on the test strip, making it possible to test thousands of indicators on one test strip.

[0034] (3) The constructed "planar structure" microchannel replaces the existing "three-dimensional structure" microchannel, which simplifies the microfluidic channel manufacturing process, reduces manufacturing costs, and allows the substrate to be made very thin, which is conducive to the popularization and promotion of microfluidic test paper chips.

[0035] (4) Compared with the prior art, by changing the capillary network diameter of the microchannel, the size of the outflow port, or setting a one-way microvalve, the flow rate and direction of the sample or reagent entering the reagent block can be precisely controlled without additional energy.

[0036] (5) Utilizing the laminar flow effect of microchannels, the position of the reagent block on the integrated test strip is set according to the target molecular weight of the indicator to be detected by the reagent block, thereby reducing the time difference of target molecules of different molecular weights in the liquid sample entering the reagent block and shortening the detection time.

[0037] (6) Compared with existing integrated test strip technology, the syringe interface, extension tube and elastic reservoir are designed to facilitate precise operation by users, avoid waste of samples or reagents and reduce the risk of aerosol contamination. Attached Figure Description

[0038] To more clearly illustrate the technical solutions of the embodiments of this application, the drawings used in the description of the embodiments will be briefly introduced below. Obviously, the drawings described below are only some embodiments of this application. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0039] Furthermore, the accompanying drawings are merely illustrative of this application and are not necessarily drawn to scale. The same reference numerals in the drawings denote the same or similar parts, and therefore repeated descriptions of them will be omitted. Some block diagrams shown in the drawings are functional entities and do not necessarily correspond to physically or logically independent entities; these functional entities can be implemented in one or more hardware modules or combinations of components.

[0040] Figure 1 This is a schematic diagram of a substrate structure according to an embodiment of this application.

[0041] Figure 2 This is a schematic diagram of a microfluidic pipeline structure according to an embodiment of this application.

[0042] Figure 3 This is a schematic diagram of a microfluidic pipeline structure including a sample orifice, according to an embodiment of this application.

[0043] Figure 4 This is a schematic diagram of a microfluidic pipeline structure including two sample orifices according to an embodiment of this application.

[0044] Figure 5A This is a schematic diagram of a microfluidic pipeline structure including a sample orifice and an extension tube, according to an embodiment of this application.

[0045] Figure 5B This is a schematic diagram of a microfluidic pipeline structure including a sample orifice, an extension tube, and a valve, according to an embodiment of this application.

[0046] Figure 6A This is a schematic diagram of a microfluidic pipeline structure including a sample hole, an extension tube, and a filter screen, according to an embodiment of this application.

[0047] Figure 6B This is a schematic diagram of a microfluidic pipeline structure including a sample hole, an extension tube, a filter screen, and a valve, according to an embodiment of this application.

[0048] Figure 7A This is a schematic diagram of a microfluidic pipeline structure including a sample orifice, an extension tube, and an elastic reservoir, according to an embodiment of this application.

[0049] Figure 7B This is a schematic diagram of a microfluidic pipeline structure including a sample orifice, an extension tube, an elastic reservoir, and a syringe interface, according to an embodiment of this application.

[0050] Figure 8 This is a schematic diagram of a microfluidic pipeline structure including a sample orifice, an extension tube, an elastic reservoir, and a filter screen, according to an embodiment of this application.

[0051] Figure 9A This is a schematic diagram of a groove unit structure according to an embodiment of this application.

[0052] Figure 9B This is a schematic diagram of another groove unit structure according to an embodiment of this application.

[0053] Figure 10A This is a schematic diagram of a reagent block structure according to an embodiment of this application.

[0054] Figure 10B This is a schematic diagram of the second reagent block structure according to an embodiment of this application.

[0055] Figure 10C This is a schematic diagram of the third reagent block structure in an embodiment of this application.

[0056] Figure 11A This is a schematic diagram of a microfluidic channel and substrate separation structure according to an embodiment of this application.

[0057] Figure 11B This is a schematic diagram of the first microfluidic channel and substrate combination structure according to an embodiment of this application.

[0058] Figure 11C This is a schematic diagram of the second type of microfluidic channel and substrate combination structure in this application embodiment.

[0059] Figure 11D This is a schematic diagram of the third type of microfluidic channel and substrate combination structure in this application.

[0060] Figure 11E This is a schematic diagram of the fourth type of microfluidic channel and substrate combination structure in this application.

[0061] Figure 11F This is a schematic diagram of the fifth type of microfluidic channel and substrate combination structure in this application.

[0062] Figure 11G This is a schematic diagram of the sixth type of microfluidic channel and substrate combination structure in this application.

[0063] Figure 11H This is a schematic diagram of another microfluidic channel and substrate separation structure according to an embodiment of this application.

[0064] Figure 12 This is a schematic diagram of a microfluidic test paper chip structure composed of a microfluidic channel, a substrate, and a reagent block, according to an embodiment of this application.

[0065] Figure 13 This is a flowchart illustrating the fabrication process of a microfluidic test paper chip according to an embodiment of this application.

[0066] Figure 14 This is a flowchart illustrating the usage of a microfluidic test paper chip according to an embodiment of this application. Detailed Implementation

[0067] To make the objectives, technical solutions, and advantages of this application clearer, the following detailed description is provided in conjunction with embodiments. It should be understood that the specific embodiments described herein are merely illustrative and not intended to limit the scope of this application.

[0068] It should be noted that the directional terms such as up, down, left, right, far, near, front, back, positive, and negative in this embodiment are only relative concepts or are based on the normal use of the product, and should not be considered as restrictive.

[0069] See Figure 1-12 One embodiment of this application provides a microfluidic test paper chip that may include a substrate 1, a microfluidic channel 2, and a reagent block 3, for multi-index detection of trace samples.

[0070] like Figure 1 As shown, in one embodiment of this application, the substrate 1 can be a rectangular strip sheet used to support the microfluidic channel 2 and the reagent block 3, and to be assembled into a complete microfluidic test paper chip. It should be noted that the substrate 1 can also be disc-shaped, square, or any other shape. The thickness, width, and length of the substrate 1 can be arbitrarily set as needed. The substrate 1 is usually made of a polymer material with good elasticity and toughness, but other materials can also be used.

[0071] See Figure 2-8 In one embodiment of this application, the microfluidic conduit 2 may include a capillary network 21, a first port 24, a groove 23 space, a second port 22, an interface 25, an extension tube 26, a filter 27, and an elastic reservoir 28.

[0072] like Figure 2 As shown, a microfluidic conduit 2 in one embodiment of this application may include a capillary network 21, which can be interconnected. A portion of the capillary network 21 may form a groove 23 space, and the capillary network 21 can communicate with the groove 23 space through a second port 22, allowing liquid flowing within the capillary network 21 to enter the groove 23 space through the second port 22. It should be noted that the capillary network 21 can be rectangular, square, circular, or any other shape, and its diameter can vary, typically between 100 micrometers and 800 micrometers. Its wall thickness typically does not exceed 500 micrometers. The capillary network 21 is typically made of a polymer material with good elasticity and toughness, but other materials can also be used. Furthermore, the groove 23 space can be rectangular, square, circular, or any other shape to match the shape and size of the reagent block 3.

[0073] like Figure 3 As shown, a microfluidic conduit 2 according to one embodiment of this application may include a capillary network 21, a first port 24, a groove space 23, a second port 22, and an interface 25. The interface 25 is used to connect a syringe. Liquid samples or reagents injected by the syringe can enter the capillary network 21 through the first port 24 via the interface 25, and flow through the capillary network 21, passing through multiple second ports 22 before entering the groove space 23.

[0074] In addition, the second port 22 may include a first-stage second port 221, a second-stage second port 222, a third-stage second port 223, and a final-stage second port 224. Among them, the first-stage second port 221 has the smallest opening size and is located in the region close to the first port 24. The flow rate of the liquid sample or reagent pushed by the syringe through the capillary network 21 and the second port 22 into the groove 23 space is the slowest. The opening sizes of the second-stage second port 222 and the third-stage second port 223 gradually increase and are located in the middle region of the capillary network 21. The flow rate of the liquid sample or reagent pushed by the syringe through the capillary network 21 and the second port 22 into the groove 23 space increases accordingly. The final-stage second port 224 has the largest opening size and is located in the region far from the first port 24. The flow rate of the liquid sample or reagent pushed by the syringe through the capillary network 21 and the second port 22 into the groove 23 space is the fastest. Thus, by using different opening sizes of the second ports 22, the liquid sample or reagent pushed by the syringe can flow from the first port 24 through the capillary network 21 into the corresponding groove 23 space almost synchronously. It should be noted that the second port 22 can be set to four size categories, or more than four size categories as needed.

[0075] In addition, the interface 25 and the first port 24 can be set at the lower end of the capillary network 21, or at the upper end, left end, or right end of the capillary network 21, or in any area between the capillary networks 21. Correspondingly, the positions of the first-level second port 221, the second-level second port 222, the third-level second port 223, and the final-level second port 224 need to be changed accordingly.

[0076] Understandably, if the liquid sample or reagent to be added needs to pass through the capillary network 21 and the second port 22 into the groove 23 space at the fastest speed, the groove 23 space can be set in the area closest to the first port 24, and this section of the capillary network 21 has the thickest diameter and the second port 22 has the largest opening; if the liquid sample or reagent to be added needs to pass through the capillary network 21 and the second port 22 into the groove 23 space at the slowest speed, the groove 23 space can be set in the area farthest from the first port 24, and this section of the capillary network 21 has the thinnest diameter and the second port 22 has the smallest opening.

[0077] like Figure 4As shown, a microfluidic conduit 2 in one embodiment of this application may include two relatively independent conduit systems, microfluidic conduit 2A and microfluidic conduit 2B, for fabricating two different types of microfluidic test strip chips. Microfluidic conduit 2A may include a capillary network 21A, a first port 24A, a groove space 23A, a second port 22A, and an interface 25A. Microfluidic conduit 2B may include a capillary network 21B, a first port 24B, a groove space 23B, a second port 22B, and an interface 25B. It is understood that the microfluidic conduit 2 may include three or more conduit systems as needed for fabricating three or more different types of microfluidic test strip chips. The shape, size, and structure of each conduit system may be the same or different. In addition, the second port 22A and the second port 22B may also be set to different size categories. The interface 25 and the first port 24 may be set at the lower end of the capillary network 21 or in any region of the capillary network 21.

[0078] like Figure 5A As shown, a microfluidic conduit 2 according to one embodiment of this application may include a capillary network 21, a first port 24, a groove 23 space, a second port 22, an interface 25, and an extension tube 26. The interface 25 is used to connect a syringe. The proximal end of the extension tube 26 is fixedly connected to the interface 25, and the distal end of the extension tube 26 is pluggably connected to the first port 24. When liquid samples or reagents need to be added, the extension tube 26 along with the interface 25 can be inserted into the first port 24 to perform the addition operation. After the addition operation is completed, the extension tube 26 along with the interface 25 can be pulled out from the first port 24. Additionally, as... Figure 5B As shown, another microfluidic conduit 2 in one embodiment of this application may include a capillary network 21, a first port 24, a groove 23 space, a second port 22, an interface 25, an extension tube 26, and a valve 241. The valve 241 is disposed near the first port 24 and is fixedly connected to the first port 24. The valve 241 is pluggably connected to the extension tube 26. During the dispensing operation, the extension tube 26 together with the interface 25 can be inserted into the proximal end of the valve 241 to open the valve 241. After the dispensing operation is completed, the valve 241 is closed, and the extension tube 26 together with the interface 25 can be pulled out from the proximal end of the valve 241 to prevent the dispensed liquid sample or reagent from spilling out.

[0079] like Figure 6A As shown, a microfluidic conduit 2 according to one embodiment of this application may include a capillary network 21, a first port 24, a groove 23 space, a second port 22, an interface 25, an extension tube 26, and a filter 27. The filter 27 is disposed between the interface 25 and the extension tube 26 for filtering large particles in liquid samples or reagents injected through the interface 25. The proximal end of the extension tube 26 is fixedly connected to the interface 25, and the distal end of the extension tube 26 is pluggably connected to the first port 24. Additionally, as... Figure 6BAs shown, another microfluidic conduit 2 in one embodiment of this application may include a capillary network 21, a first port 24, a groove 23 space, a second port 22, an interface 25, an extension tube 26, and a valve 241. The valve 241 is disposed near the first port 24 and is fixedly connected to the first port 24. The valve 241 is pluggable and detachable from the extension tube 26. The valve 241 is used to prevent the added liquid sample or reagent from spilling.

[0080] like Figure 7A As shown, a microfluidic conduit 2 in one embodiment of this application may include a capillary network 21, a first port 24, a groove 23 space, a second port 22, an extension tube 26, and an elastic reservoir 28. The elastic reservoir 28 is connected to the extension tube 26, and the extension tube 26 is pluggably connected to the first port 24. The elastic reservoir 28 may include a reservoir body 281 and an injection vessel 282. Liquid samples or reagents to be injected can be inserted into the injection vessel 282 through a syringe needle and injected into the reservoir body 281. After injection, the syringe needle can be withdrawn. The liquid sample or reagent can be slowly and continuously injected into the capillary network 21 under the elastic recoil force of the reservoir body 281. It should be noted that the volume, elasticity, and material of the reservoir body 281 can be set as needed, and the shape and size of the injection vessel 282 can be set as needed. Figure 7B As shown, another microfluidic conduit 2 in one embodiment of this application may include a capillary network 21, a first port 24, a groove space 23, a second port 22, an extension tube 26, an elastic reservoir 28, and an interface 25. The elastic reservoir 28 may include a reservoir body 281, and the interface 25 can be directly connected to a syringe. It should be noted that cell lysis buffer can be pre-coated inside the elastic reservoir 28 as needed for the detection of samples that may contain blood or tissue components, such as urine, gastric juice, and fecal filtrate.

[0081] like Figure 8 As shown, a microfluidic conduit 2 in one embodiment of this application may include a capillary network 21, a first port 24, a groove 23 space, a second port 22, an extension tube 26, an elastic reservoir 28, and a filter 27. The elastic reservoir 28 is connected to the extension tube 26, and the extension tube 26 is pluggably connected to the first port 24. The elastic reservoir 28 may include a reservoir body 281 and a syringe 282. The filter 27 is disposed at the tail end of the elastic reservoir 28 and is connected to the extension tube 26. Liquid samples or reagents to be added can be injected into the reservoir 282 through a syringe needle and then into the reservoir body 281. When the liquid sample or reagent is slowly and continuously injected into the capillary network 21 under the elastic recoil force of the reservoir body 281, the filter 27 filters out large particles, preventing large particles from entering the capillary network 21. It should be noted that the syringe 282 can be replaced by an interface 25, which can be directly connected to a syringe.

[0082] like Figure 9A As shown, in one embodiment of this application, the groove 23 space unit of the microfluidic conduit 2 may include a surrounding capillary network 21, a first port 24, and a second port 22. The capillary network 21, with its walls, forms a enclosure for the groove 23 space. Liquid samples or reagents can enter the capillary network 21 through the first port 24 and enter the groove 23 space through the second port 22. Additionally, as... Figure 9B As shown, in another embodiment of this application, the groove 23 space unit of the microfluidic pipeline 2 may include a surrounding capillary network 21, a first port 24, a second port 22, and a microvalve 29. The capillary network 21 forms a fence around the groove 23 space with its walls. The microvalve 29 is a one-way valve. Liquid samples or reagents can enter the capillary network 21 through the first port 24 and enter the groove 23 space through the second port 22 and the microvalve 29. The microvalve 29 can prevent the liquid in the groove 23 space from flowing back into the capillary network 21.

[0083] like Figure 10A As shown, the reagent block 3 in this embodiment can be a disk-shaped structure. The types of reagent block 3 include, but are not limited to, dry chemistry detection reagent blocks, immunological detection reagent blocks, and chip reagent blocks. Reagent block 3 can include single-item detection reagent combinations and multi-item detection reagent combinations. Additionally, as... Figure 10B As shown, another reagent block 3 in this embodiment may include a reaction component 32 and a waste liquid absorption component 31. The reaction component 32 is used to perform a colorimetric reaction, and the waste liquid absorption component 31 is used to absorb excess liquid sample, reagent, or waste liquid from the colorimetric reaction process. Furthermore, as... Figure 10C As shown, another reagent block 3 in the application embodiment may include a reaction component 32, a waste liquid absorption component 31, and a filter membrane component 33. The filter membrane component 33 is disposed between the reaction component 32 and the second port 22, and is used to filter large particulate components in the detection sample. It is understood that the reagent block 3 may be a dry test paper block that adsorbs colorimetric reagents, a semi-dry test paper block or a gel block that adsorbs colorimetric reagents, or a detection unit composed of one or more micro-chambers pre-coated with colorimetric reagents.

[0084] like Figure 11A As shown, in one embodiment of this application, the substrate 1 can be bonded and combined with a microfluidic channel 2 to obtain the following: Figure 11BThe illustration shows a substrate 1 microfluidic channel 2 assembly, wherein the substrate 1 and the microfluidic channel 2 need to be matched in size, shape, and material. A capillary network 21, with its walls forming a groove 23, is combined with the substrate provided by the substrate 1 to form the groove 23. The interface 25 of the substrate 1 microfluidic channel 2 assembly is typically located on the side edge of the substrate 1 microfluidic channel 2 assembly to facilitate injection operations. Furthermore, the substrate 1 can be bonded to different types of microfluidic channels 2 to form different types of substrate 1 microfluidic channel 2 assemblies to meet specific needs, such as... Figure 11C The substrate 1 and microfluidic channel 2 assembly shown may include a substrate 1 and two sets of microfluidic channel 2 systems, such as Figure 11D The substrate 1 and microfluidic channel 2 assembly shown may include a substrate 1 and a microfluidic channel 2 with an extension tube 26, such as Figure 11E The microfluidic conduit 2 assembly shown may include a substrate 1 and a microfluidic conduit 2 with an extension tube 26 and a filter screen 27, such as Figure 11F The microfluidic conduit 2 assembly shown may include a substrate 1 and a microfluidic conduit 2 with an extension tube 26, a capsule 281, and an injection vessel 282, as shown. Figure 11G The microfluidic conduit 2 assembly shown may include a substrate 1 and a microfluidic conduit 2 with an extension tube 26, a filter 27, a capsule 281, and an injection vessel 282.

[0085] In addition, such as Figure 11H As shown, in one embodiment of this application, a substrate 1 can be bonded to two microfluidic channels 2 to form a substrate 1-microfluidic channel 2 assembly. The substrate 1 and the two microfluidic channels 2 need to be compatible in size, shape, and material. The two microfluidic channels 2 are bonded to the front and back sides of the substrate 1 respectively, which can increase the number of grooves 23. It should be noted that the two microfluidic channels 2 can be of the same type or different types.

[0086] like Figure 12 As shown, the microfluidic test paper chip of this application embodiment may include a substrate 1, a microfluidic channel 2, and a reagent block 3. The reagent block 3 can be printed into the dot-matrix arrangement of grooves 23 formed by the substrate 1 and the microfluidic channel 2. A syringe connection interface 25 is used to push liquid samples or reagents into the capillary network 21 through the first port 24, where they flow and enter the grooves 23 through the second port 22. The waste liquid adsorption component 31 of the reagent block 3 in the grooves 23 has a capillary adsorption function, attracting liquid samples or reagents to permeate and wet the reaction component 32 of the reagent block 3. When the reaction component 32 of the reagent block 3 is saturated, the capillary adsorption function of the waste liquid adsorption component 31 weakens or disappears, and the permeation and wetting of the liquid sample or reagent ends.

[0087] It should be noted that the dot-matrix arrangement of the grooves 23 of the microfluidic test paper chip substrate 1 and microfluidic channel 2 in this embodiment can be divided into multiple regions, based on the distance between the grooves 23 and the first port 24. Similarly, the reagent blocks 3 can be divided into multiple categories, based on the size of the target molecular weight of the index detected by the reagent block 3. If the target molecular weight of the index detected by the reagent block 3 is large, the reagent block 3 is placed in the groove 23 region close to the first port 24; if the target molecular weight of the index detected by the reagent block 3 is small, the reagent block is placed in the groove 23 region far from the first port 24. Thus, by utilizing the laminar flow effect of the microfluidic channel 2, different target molecules in the liquid sample enter different categories of reagent blocks 3 almost simultaneously, shortening the detection time and improving detection efficiency.

[0088] It should be noted that the amount of liquid sample or reagent expected to be required by the microfluidic test paper chip in this application embodiment can be precisely designed and obtained from actual testing, thereby providing a reference for users.

[0089] It should be noted that the microfluidic test paper chip reagent block 3 in this application embodiment can also be divided into multiple categories according to different color reaction mechanisms. Reagent blocks 3 with the same or similar mechanisms can be set in the same area, and the same set of microfluidic channels 2 can be set in the same area to facilitate the user to add liquid samples or reagents.

[0090] See Figure 13 The microfluidic test paper chip preparation method process 100 of this application embodiment may include the following steps.

[0091] 110. Design a microfluidic test paper chip, including microfluidic channel wiring and reagent block layout.

[0092] First, determine the categories, testing indicators, and performance indicators of the microfluidic test paper chip to meet user needs.

[0093] Then, using design software, a microfluidic pipeline diagram is drawn. The microfluidic pipeline diagram includes at least one or more capillary networks, one or more first ports, multiple recessed spaces, multiple second ports, and one or more interfaces. It may also include one or more extension tubes, one or more filters, and one or more elastic reservoirs.

[0094] The reagent blocks can be divided into multiple groove regions based on their distance from the first port. Similarly, they can be divided into multiple groove regions to accommodate different reagent blocks based on the target molecular weight of the reagent block. This determines the reagent block layout scheme. Furthermore, reagent blocks can be categorized according to their different colorimetric reaction mechanisms. Reagent blocks with the same or similar mechanisms can be placed in the same groove region, and the same set of microfluidic channels can be installed in that region for convenient addition of liquid samples or reagents.

[0095] The substrate is designed based on the microfluidic pipeline circuit diagram.

[0096] The calculation of the amount of liquid sample or reagent required for microfluidic test paper chip detection provides a reference for users.

[0097] 120. Microfabrication of microfluidic channels. Typically, polymer or silicon-based materials are chosen, and injection molding, etching, or 3D printing techniques are used, based on the microfluidic channel design from step 110. Figure 3 3D modeling was used to create microfluidic channels.

[0098] 130. Microfabrication of reagent blocks. Reagent blocks are typically disc-shaped dry or semi-dry test strips, gel blocks, or one or more micro-chambers forming a detection unit. Types of reagent blocks include dry chemical detection reagent blocks, immunological detection reagent blocks, and chip reagent blocks.

[0099] 140. Microfluidic channels are bonded to the substrate, and capillary networks form a grooved lattice on the substrate. The microfluidic channels fabricated in step 120 are bonded to the matching substrate by adhesive or thermal bonding. A substrate can have one or more sets of microfluidic channels bonded to one side, or two or more sets of microfluidic channels bonded to both the front and back sides. It is understood that if 3D printing technology is used to fabricate the microfluidic channels, a microfluidic channel-substrate composite can be created.

[0100] 150. The reagent blocks are printed onto the substrate groove matrix, using existing technology (Patent Publication No.: CN112362648A). It should be noted that the semi-dry reagent blocks, gel reagent blocks, or liquid reagent blocks can be covered with a micro-capsule or film.

[0101] 160. Install at least one or more components, including interface, extension tube, valve, elastic reservoir, and filter, print identification code in blank area of ​​substrate, produce complete microfluidic test paper chip, and place in packaging box.

[0102] See Figure 14 The microfluidic test paper chip usage process 200 of this application embodiment may include the following steps.

[0103] 210. Select a microfluidic detection test strip chip. Depending on the sample type and testing purpose, select one microfluidic detection test strip chip or a combination of multiple microfluidic detection test strip chips for sample testing.

[0104] 220. Sample Addition. Connect the sample addition assembly to the microfluidic test paper chip. Draw up the liquid sample with the syringe, connect it to the microfluidic test paper chip interface, and the sample can be added directly. If the sample is a solid or semi-solid specimen, such as dried or formed feces, dried blood, or urine residue, it needs to be dissolved with physiological saline or pure water before drawing up the sample with the syringe, connecting it to the microfluidic test paper chip interface, and adding the sample. It should be noted that the liquid sample volume needs to reach the minimum amount indicated on the microfluidic test paper chip to ensure that each reagent block of the microfluidic test paper chip is fully wetted. If the liquid sample volume is insufficient, the sample can be diluted appropriately to reach the minimum amount indicated on the microfluidic test paper chip.

[0105] 230. Add reagents. Following the instructions for use of the microfluidic test paper chip, before or after sample addition, draw a specific dose of one or more reagents or a combination of reagents using a syringe, connect the syringe to the microfluidic test paper chip interface, and add the reagents.

[0106] 240. Control the colorimetric reaction conditions. Follow the instructions for use of the microfluidic test paper chip to provide a suitable temperature and humidity environment. Remove the micro-cover plate or film covering the reagent block and allow an appropriate reaction time.

[0107] 250. Scan and detect, obtain results. After the color reaction in step 240 is completed, the microfluidic test paper chip is scanned by a photosensitive sensor under appropriate incident light conditions to obtain the color reaction data of each reagent block of the microfluidic test paper chip. The algorithm calculates the detection data of each reagent block of the microfluidic test paper chip, thereby obtaining the detection results of multiple indicators in the sample.

[0108] The above description is merely an embodiment of this application and does not constitute any limitation on the technical scope of this application. Therefore, any minor modifications, equivalent changes, and alterations made to the above embodiments based on the technical essence of this application shall still fall within the scope of the technical solution of this application. Those skilled in the art should recognize that different methods can be used to implement the described functions for each specific application, but such implementation should not be considered beyond the scope of this application.

Claims

1. A microfluidic test paper chip for multi-index detection of trace samples, characterized in that, include: substrate; A microfluidic channel, bonded to a substrate surface, is used to control the flow rate and direction of liquid samples or reagents. The microfluidic channel includes a sample dispensing component, at least one first port, at least one capillary network, and multiple second ports. The sample dispensing component is connected to the first port, the first port is connected to the capillary network, and the capillary network is connected to the second ports. The capillary network forms multiple grooves arranged in a dot matrix on the substrate. Each groove is connected to the capillary network through a second port. The second port includes a primary second port, a secondary second port, and a final second port. The primary second port has the smallest opening and is used to connect the capillary network to the groove near the first port. The final second port has the largest opening and is used to connect the capillary network to the groove away from the first port. A reagent block is disposed within a groove formed by a substrate and a microfluidic channel. The reagent block includes a reaction component and a waste liquid absorption component, and is used to perform a colorimetric reaction. The placement of the reagent block within the groove formed by the substrate and the microfluidic channel involves determining the position of the reagent block in the groove on the substrate based on the target molecular weight of the reagent block. Specifically, reagent blocks with a larger target molecular weight are placed in the groove closer to the first port, while reagent blocks with a smaller target molecular weight are placed in the groove farther away from the first port.

2. The microfluidic test paper chip as described in claim 1, characterized in that, The sample dispensing assembly includes at least one sample orifice, which includes a first interface for connecting a syringe to dispense a liquid sample.

3. The microfluidic test paper chip as described in claim 2, characterized in that, The sample loading assembly includes an extension tube, the proximal end of which is connected to a first interface, and the distal end of which is pluggably connected to a first port.

4. The microfluidic test paper chip as described in claim 2, characterized in that, The sample dispensing assembly includes at least one reagent port, which includes a second interface and a second extension tube for connecting a syringe and dispensing reagents. The proximal end of the second extension tube is connected to the second interface, and the distal end is pluggably connected to the first port.

5. The microfluidic test paper chip as described in claim 2 or 4, characterized in that, The sample dispensing assembly includes at least one elastic reservoir for storing liquid samples or reagents and slowly and continuously injecting the liquid samples or reagents into the capillary network through the first port. The elastic reservoir includes an injection vessel, a vessel body, and a valve. The injection vessel is used to connect to the syringe needle, the injection vessel is connected to the vessel body, and the vessel body is pluggably connected to the first port through the valve.

6. The microfluidic test paper chip as described in claim 1, characterized in that, The microfluidic pipeline includes multiple microvalves, which are located between the second port and the groove. These microvalves are one-way valves used to control the flow direction of liquid samples or reagents.

7. The microfluidic test paper chip as described in claim 1, characterized in that, The reagent block includes a filter membrane component, which is disposed between the reaction component and the second port for filtering large particulate components in the detection sample.

8. A method for preparing a microfluidic test paper chip as described in any one of claims 1-7, characterized in that, The preparation method includes the following steps: Step 1: Design the microfluidic test paper chip, including microfluidic channel wiring and reagent block layout; Step 2: Microfabrication to create microfluidic channels and reagent blocks; Step 3: Microfluidic channels are bonded to the substrate, and capillary networks form a groove lattice on the substrate; Step 4: Print reagent blocks onto the substrate groove matrix; Step 5: Install the extension tube and elastic reservoir, print the identification code, and place it in the packaging box.

9. A method of using a microfluidic test paper chip as described in any one of claims 1-7, characterized in that, The usage includes the following steps: Step 1: Select the microfluidic test paper chip; Step 2, add the sample; Step 3, add reagents; Step 4: Control the colorimetric reaction conditions; Step 5: Scan and obtain the test results.

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