1. A microfluidic control cartridge, comprising: the method comprises the following steps: the detection device comprises a card shell upper cover (100), a card shell lower cover (200), a detection membrane (300), a micro-fluidic plate (400) with waterproofness and water absorption paper (500), wherein the card shell upper cover (100) is detachably connected with the card shell lower cover (200), a window (302) used for being embedded into the detection membrane (300) is arranged on the card shell upper cover (100), the edge of the window (302) and the edge of the card shell upper cover (100) form a slope, the window (302) is downwards sunken, the detection membrane (300) is fixedly connected onto the window (302) through four small convex columns (301) with the downwards bottoms of the window (302), the window (302) is arranged on the upper side of the micro-fluidic plate (400), the micro-fluidic plate (400) is slidably arranged between the card shell upper cover (100) and the card shell lower cover (200), and the detection membrane (300) is arranged on the lower side of the window (302) of the card shell upper cover (100), the water absorption paper (500) is positioned on one side, close to the lower card shell cover (200), of the microfluidic board (400), the lower card shell cover (200) is used for containing water absorption materials, and the microfluidic board (400) moves between the upper card shell cover (100) and the lower card shell cover (200) so that the liquid permeation chip is absorbed by the water absorption paper (500) or the water absorption materials.

2. A microfluidic control cartridge according to claim 1, wherein: the detection membrane (300) is a sheet-shaped supporting material which can seep water and can adsorb protein.

3. A microfluidic control cartridge according to claim 2, wherein: the detection membrane can adsorb protein, and different samples can be fixed on the detection membrane by using a chip instrument for bearing a protein array (310) for realizing high-throughput detection.

4. A microfluidic control cartridge according to claim 3, wherein: the protein arrays (310) are uniformly distributed on one side of the nitrocellulose membrane close to the upper cover (100) of the card shell at equal intervals.

5. A microfluidic control cartridge according to claim 3, wherein: the protein array (310) is irregularly arranged on one side of the nitrocellulose membrane close to the upper cover (100) of the card shell.

6. A microfluidic control cartridge according to claim 1, wherein: the lower clamping shell cover (200) and the upper clamping shell cover (100) are provided with track grooves (210) for the movement of the microfluidic plate (400), the thickness of the track grooves (210) is the same as that of the microfluidic plate, and the microfluidic plate (400) can move in the track grooves (210) to realize fluid control.

7. A microfluidic control cartridge according to claim 6, wherein: the micro-fluidic plate (400) comprises a control part (410) for controlling the micro-fluidic plate (400), a limiting part (420) for limiting the movement position of the micro-fluidic plate (400), and a baffle part (430) for preventing liquid from flowing out, wherein the control part (410), the baffle part (430) and the limiting part (420) are fixedly connected.

8. A microfluidic control cartridge according to claim 7, wherein: the cross section area of the limiting part (420) and the attached micro-fluidic plate (400) is larger than that of the cross section of the micro-fluidic plate (400).

9. A microfluidic control cartridge according to claim 8, wherein: the control part (410) protrudes outwards from the baffle part (430) and is suitable for fingers to hold.

10. A microfluidic control cartridge according to claim 9, wherein: a cavity for bearing liquid can be formed between the microfluidic board (400) and the upper cover (100) of the card shell in a closed mode.

Background

The immunoassay method has the advantages that the immunoassay method has multiple operation steps, each step of reaction needs to ensure the reaction time, non-reaction interferents need to be washed and removed to carry out the next reaction, and a clinical sample has potential biological pollution. The existing immunoassay reaction device is difficult to detect at high flux, and has large requirement on clinical specimens and large wound on patients.

The most similar realization scheme of the prior invention is a 96-hole enzyme-linked immunosorbent plate, but the prior realization scheme does not have a liquid adsorption device, needs manual plate washing, is easy to pollute the environment and is difficult to realize trace high-flux detection.

Another implementation is a colloidal gold chip that percolates the cartridge, but it does not control the retention time of the fluid reacting with the chip and therefore does not allow sensitive detection.

Meanwhile, a 96-well polystyrene plate is generally used in the existing immunoassay technology, the antigen or antibody is coated in the micropore by utilizing the adsorption of polystyrene to protein, the liquid to be detected containing the antigen and the antibody is added into the micropore for incubation reaction during the reaction, the liquid is manually removed and washed after the reaction is finished, then the secondary antibody or protein which contains a marker and can react with the protein to be detected is added, the incubation reaction is carried out for a certain time, the residual liquid is manually removed and washed after the reaction is finished, then a chromogenic substrate is added, the reaction is developed, the value is read, multiple rounds of incubation and washing processes are required, the operation steps are complex, and the waste liquid is easy to pollute the environment. The detection flux is low, if the flux needs to be improved, only parallel experiments can be carried out, and the requirement on the amount of the sample is high.

Therefore, a new microfluidic control cartridge needs to be designed to solve the above technical problems.

Disclosure of Invention

The technical problem to be solved by the invention is to overcome the defects of the prior art, and to solve the technical problem, the invention adopts the basic concept of the technical scheme that: the utility model provides a micro-fluidic control card shell, includes card shell upper cover, card shell lower cover, permeable liquid's detection membrane, micro-fluidic board, the paper that absorbs water that has the water proof, card shell upper cover with card shell lower cover can dismantle the connection, card shell upper cover is last to be offered and is used for the embedding detect the downwards sunken window of membrane, the detection membrane is with four little projection fixed connection in the card shell upper cover, the sliding tray setting of micro-fluidic board the card shell upper cover with between the card shell lower cover, the shape of sliding tray is the U type, and when micro-fluidic board inserted, the three limit of micro-fluidic board can the water proof with the tight real laminating of sliding tray, and baffle portion just blocks in the space that upper and lower card shell formed for the micro-fluidic board motion. Therefore, the upper cartridge and the microfluidic plate can form a cup shape for carrying the reaction liquid. The water absorbing paper is positioned on one side of the microfluidic control plate close to the lower cover of the card shell, the lower cover of the card shell is used for containing water absorbing materials, and the water absorbing paper covers the uppermost side of the microfluidic control plate or is used as the whole. The micro-fluidic plate moves between the detection membrane and the absorbent paper.

In order to greatly improve the detection effect of the detection membrane, the detection membrane is preferably a nitrocellulose membrane as the microfluidic control card shell.

In order to greatly improve the high-throughput detection effect of the protein array, as a preferable microfluidic control card shell of the invention, the nitrocellulose membrane can be stamped or sprayed with a plurality of protein arrays for realizing high-throughput detection.

In order to greatly improve the high-flux detection effect of the protein array, the microfluidic control card shell is preferably used, and a small convex column is arranged below a window of an upper cover of the card shell, so that a detection membrane can be fixed, and the positioning effect is achieved.

In order to greatly improve the stability and detection effect of the protein array, the protein array is preferably uniformly distributed on one side of the nitrocellulose membrane close to the upper cover of the card shell at equal intervals.

In order to greatly improve the stability and detection effect of the protein array, the microfluidic control card shell is preferably used, and the protein spots are arranged on one side of the nitrocellulose membrane close to the upper cover of the card shell in an array manner.

In order to greatly improve the limiting capacity of the track groove on the microfluidic control plate, the microfluidic control card shell is preferably provided with the track groove which is convenient for the movement of the microfluidic control plate between the upper cover of the card shell and the lower cover of the card shell.

In order to greatly improve the control and operation capabilities of the microfluidic control card shell, the microfluidic control card shell is preferably used as a microfluidic control card shell of the invention, the microfluidic control card shell comprises a control part for controlling the microfluidic control plate, a limiting part for limiting the movement position of the microfluidic control plate, and a baffle part for preventing liquid from flowing out, and the control part, the baffle part and the limiting part are fixedly connected.

In order to greatly improve the limiting capacity between the limiting part and the track groove, the cross section area of the limiting part and the attached microfluidic plate is preferably larger than that of the microfluidic plate.

In order to greatly control the distance that the microfluidic control plate is pulled out and enable liquid to be easily absorbed by absorbent paper below the window, as an optimal preference of the microfluidic control card shell, the limiting part is positioned at a position close to the top end of the microfluidic control plate, and the width of the limiting part is smaller than the distance from the edge of the window of the upper cover of the card shell to the edge of the corresponding upper cover.

In order to greatly improve the tightness of the microfluidic control plate, the microfluidic control plate is preferably designed with a baffle part as a preferred microfluidic control card shell of the invention, and the baffle part is used for enabling the microfluidic control plate and the upper card shell to form a closed bearing cavity when the microfluidic control plate is pushed in.

In order to greatly improve the overall stability between the control part and the limiting part, the control part is preferably protruded outwards from the baffle part, so that the microfluidic control card shell is suitable for being held by fingers and is convenient to operate and control.

In order to greatly improve the liquid bearing capacity, it is preferable that, as a microfluidic control cartridge of the present invention, a cavity for bearing liquid is formed between the microfluidic plate and the cartridge upper cover.

After the technical scheme is adopted, compared with the prior art, the invention has the following beneficial effects.

The invention provides a microfluidic control card shell which comprises a card shell upper cover, a card shell lower cover, a detection membrane, a microfluidic control plate with water-resisting property and water-absorbing paper, wherein the microfluidic control plate moves in a track groove between the card shell upper cover and the card shell lower cover so that a liquid permeating chip is absorbed by the water-absorbing paper. The protein loaded on the chip can fully react with various antigens or antibodies and chromogenic substrates step by step. The protein loaded on the chip can be various and can be arranged in a microarray form. The reaction time of the micro-fluid and the protein chip can be controlled by the micro-fluidic plate, and the operation steps of immunoreaction, washing, color development and the like are realized by the micro-fluidic plate and the absorbent paper under the plate.

Drawings

FIG. 1 is a schematic structural diagram of a microfluidic control cartridge according to the present invention;

FIG. 2 is a schematic structural diagram of an embodiment of a microfluidic control cartridge according to the present invention;

FIG. 3 is an analytical representation of a microfluidic control cartridge according to the present invention;

in the figure, 100, a card shell upper cover; 200. a lower cover of the clamping shell; 210. a track groove; 300. a detection membrane; 301. a small convex column; 302. a window; 310. a protein array; 400. a microfluidic plate; 410. a control unit; 420. a limiting part; 430. a baffle portion; 500. absorbent paper.

Detailed Description

In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention is described in further detail below with reference to the accompanying drawings and embodiments. It should be understood that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention.

In the description of the present invention, it is to be understood that the terms "length", "width", "upper", "lower", "front", "rear", "left", "right", "vertical", "horizontal", "top", "bottom", "inner", "outer", and the like, indicate orientations or positional relationships based on the orientations or positional relationships illustrated in the drawings, and are used merely for convenience in describing the present invention and for simplicity in description, and do not indicate or imply that the devices or elements referred to must have a particular orientation, be constructed in a particular orientation, and be operated, and thus, are not to be construed as limiting the present invention. Further, in the description of the present invention, "a plurality" means two or more unless specifically defined otherwise.

Referring to fig. 1 to 3, the present invention provides a technical solution: a microfluidic control card shell comprises an upper card shell cover 100, a lower card shell cover 200, a detection membrane 300, a microfluidic board 400 with waterproofness and absorbent paper 500, wherein the upper card shell cover 100 is detachably connected with the lower card shell cover 200, the upper card shell cover is provided with a window for embedding the detection membrane, a small convex column 301 is arranged below the window 302 of the upper card shell cover and can fix the detection membrane and achieve the positioning effect, the detection membrane 300 is fixedly connected below the upper card shell cover window 302 through the small convex column 301, a microfluidic board 400 sliding rail is arranged between the upper card shell cover 100 and the lower card shell cover 200, the detection membrane 300 is arranged between the upper card shell cover window 302 and the microfluidic board 400, the absorbent paper 500 is attached to one side of the microfluidic board 400 close to the lower card shell cover 200, the lower card shell cover 200 is used for containing absorbent materials, and the board 400 moves between the upper card shell cover 100 and the lower card shell cover 200, so that the liquid permeating core sheet is absorbed by the water absorbent paper 500.

Example 1: antigens of bacteria and viruses commonly infected with the respiratory tract were spotted in the form of a microarray on a 1 cm square nitrocellulose membrane: influenza b virus, syncytial virus, adenovirus, mycoplasma pneumoniae antigen, legionella, Q fever rickettsia, chlamydia pneumoniae, influenza a virus, parainfluenza virus, 9 antigens, and then blocking the uncoated site with PBS containing 10% of fire-extinguishing bovine serum. After drying, the human serum IgG detection card is assembled on the lower side of the window 302 of the microfluidic card shell upper cover 100 to prepare the human serum IgG detection card with nine respiratory tracts. For the assay, the patient's serum was mixed with PBS at a ratio of 1: diluting 10, adding 200 microliters of diluted serum into a depression formed at the window 302 of the upper cover of the microfluidic detection card, allowing the serum to react with the chip for 30 minutes, pulling out the microfluidic plate 400, allowing the reaction liquid to be absorbed by the absorbent paper 500 below the microfluidic plate 400, adding 200 microliters of PBS, washing, and absorbing by the absorbent paper 500; the microfluidic plate 400 was inserted, goat anti-human IgG polyclonal antibody reaction solution labeled with colloidal gold was added to the upper cap of the chip to allow the gold solution to react with the antigen-antibody complex on the chip for 30 minutes, the microfluidic plate 400 was pulled out, unreacted colloidal gold solution was absorbed by the absorbent paper 500 stored in the lower cap 200 of the card housing, then 200 microliters of PBS containing 10% BSA was added, and the washing solution was absorbed by the absorbent paper 500 after washing. The NC membrane window of the microfluidic card shell upper cover 100 can show colloidal gold red spots in positive and no spots in negative according to the reaction condition. The detection of 9 virus IgG antibodies can be completed by using 20 microliter serum once, and the operation is simple and convenient, and the sensitivity is not lower than that of ELISA.

Example 2: preparing and screening specific respiratory tract anti-influenza B virus, syncytial virus, adenovirus, mycoplasma pneumoniae antigen, legionella, Q fever rickettsia, chlamydia pneumoniae, influenza A virus and parainfluenza virus, and detecting the antigen by using a double-antibody sandwich method.

The 9 respiratory antigen-corresponding antibodies were spotted in a microarray format on nitrocellulose membranes, and the uncoated sites were blocked with PBS containing 10% of fire-fighting bovine serum. And after drying, assembling the protein into a microfluidic card shell to prepare the nine-respiratory-tract antigen detection card. When in detection, a throat swab of a patient is diluted to 200 microliters by PBS containing 0.05 percent of Tween-20, the diluted throat swab is added into a concave part formed at the window 302 by the upper cover of the microfluidic detection card, a sample and a chip react for 30 minutes, the microfluidic plate 400 is pulled out, the reaction liquid is absorbed by the absorbent paper 500 below the microfluidic plate 400, and then 200 microliters of PBS are added, washed and absorbed by the absorbent paper 500; inserting the micro-fluidic plate 400, adding 300 microliters of colloidal gold mixed solution labeled by 9 different mouse anti-respiratory antigen monoclonal antibodies into the upper cover of the chip, reacting the gold solution with the antigen-antibody complex on the chip for 30 minutes, pulling out the micro-fluidic plate 400, absorbing unreacted colloidal gold solution by the absorbent paper 500 stored in the lower cover 200 of the card shell, then adding 200 microliters of PBS containing 10% BSA, washing, and absorbing the washing solution by the absorbent paper 500. The NC membrane window of the microfluidic card shell upper cover 100 can show colloidal gold red spots in positive and no spots in negative according to the reaction condition. The detection of 9 viruses can be completed by using one throat swab sample at one time, and the method is simple and convenient to operate, and the sensitivity is not lower than that of ELISA.

Example 3: antigens of bacteria and viruses commonly infected with the respiratory tract were spotted on nitrocellulose membranes in the form of microarrays: influenza b virus, syncytial virus, adenovirus, mycoplasma pneumoniae antigen, legionella, Q fever rickettsia, chlamydia pneumoniae, influenza a virus, parainfluenza virus, 9 antigens, and then blocking the uncoated site with PBS containing 10% of fire-extinguishing bovine serum. And after drying, assembling the human serum IgM detection card into a microfluidic card shell to prepare the human serum IgM detection card for nine respiratory tracts. For testing, the microfluidic plate 400 is inserted and the patient's serum is mixed with PBS in a 1: 10 to 200 microliter of liquid, adding the liquid into a depression formed at a window of an upper cover of the microfluidic detection card, allowing the serum to react with the chip for 30 minutes, pulling out the microfluidic plate 400, allowing the reaction liquid to be absorbed by the absorbent paper 500 below the microfluidic plate 400, adding 200 microliter of PBS, washing for three times, and absorbing by the absorbent paper 500; inserting the microfluidic plate 400, adding a goat anti-human IgM polyclonal antibody marked by horseradish peroxidase into the upper cover of the chip, allowing the secondary antibody to react with the antigen-antibody complex on the chip for 30 minutes, pulling out the microfluidic plate 400, absorbing the unreacted secondary antibody liquid by the absorbent paper 500 stored in the lower cover 200 of the card shell, then adding 200 microliters of PBS containing 10% BSA, washing for three times, and absorbing the washing liquid by the absorbent paper 500. Inserting the microfluidic plate 400, adding DAB substrate solution, controlling the color reaction time, pulling the microfluidic plate 400 after the color reaction is finished, and sucking away the redundant liquid by the absorbent paper 500. The NC membrane window of the microfluidic card shell upper cover 100 can be used for generating brown spots in positive and generating no spots in negative according to the reaction condition. The detection of 9 virus IgM antibodies can be completed by using 20 microliter serum once, and the operation is simple and convenient, and the sensitivity is not lower than ELISA.

As a technical optimization scheme of the present invention, the detection membrane 300 is a nitrocellulose membrane.

Meanwhile, the window 302 of the upper cover 100 of the card housing is lower than the upper frame and higher than the lower frame of the upper cover 100 of the card housing, the detection membrane 300 can be embedded into the window 302, and the four walls of the upper cover 100 of the card housing and the microfluidic board 400 can form a closed cup shape for carrying liquid.

In this embodiment: the application of the nitrocellulose membrane can improve the detection effect of the detection membrane 300.

As a technical optimization scheme of the invention, a plurality of protein arrays 310 for realizing high-throughput detection can be arranged on the nitrocellulose membrane.

In this embodiment: the arrangement of multiple protein arrays 310 can further improve the high-throughput detection effect of the protein arrays 310.

As a technical optimization scheme of the invention, the protein arrays 310 are uniformly arranged on one side of the nitrocellulose membrane close to the upper cover 100 of the card shell at equal intervals.

In this embodiment: the protein array 310 with equal spacing can improve the stability and detection effect of the protein array 310.

As a technical optimization scheme of the invention, the protein arrays 310 are irregularly arranged on one side of the nitrocellulose membrane close to the upper cover 100 of the card shell.

In this embodiment: the irregular shape of the protein array 310 can improve the stability and detection effect of the protein array 310.

As a technical optimization scheme of the present invention, the upper card housing cover 100 and the lower card housing cover 200 are provided with track grooves 210 for preventing the microfluidic board 400 from moving out.

In this embodiment: the track groove 210 can improve the position-limiting capability of the microfluidic plate 400.

As a technical optimization scheme of the present invention, the microfluidic plate 400 includes a control part 410 for controlling the microfluidic plate 400 and a position limiting part 420 for limiting a movement position of the microfluidic plate 400, and the control part 410 and the position limiting part 420 are fixedly connected.

In this embodiment: the control portion 410 and the position-limiting portion 420 can be matched with each other to improve the control and operation capability of the microfluidic plate 400.

As a technical optimization scheme of the present invention, the cross-sectional area of the limiting part 420 plus the micro-fluidic plate attached thereto is larger than the cross-sectional area of the micro-fluidic plate.

In this embodiment: the size of the obstructed position and the limiting part 420 and the rail groove 210 can improve the limiting capacity of the microfluidic board 400, the microfluidic board 400 is similar to a drawer without a four-sided board, the cross section of the microfluidic board attached to the limiting part 420 is larger than that of the microfluidic board 400, the redundant area is below the microfluidic board 400 to prevent the microfluidic board 400 from sliding out of a slide rail of a card shell, meanwhile, the limiting part 420 is positioned at a position close to the top end of the microfluidic board 400, and the width of the limiting part is smaller than the horizontal distance from the edge of a window to the edge of a corresponding upper cover to ensure that the microfluidic board 400 can be completely pulled out and avoid the position of the window 302 to avoid influencing the absorption of liquid of the window 302.

As a technical optimization of the present invention, the control portion 410 is protruded outward from the baffle portion 430, which is suitable for being held by fingers.

In this embodiment: the control part 410 and the position-limiting part 420 having different shapes can improve the overall stability of the movement of the microfluidic plate 400.

As a technical optimization scheme of the present invention, a cavity for carrying liquid is formed between the microfluidic board 400 and the four walls of the upper cover 100 of the cartridge.

In this embodiment: the cavity can improve the liquid bearing capacity of the invention.

The working principle is as follows:

the invention provides a microfluidic control card shell which comprises a card shell upper cover 100, a card shell lower cover 200, a detection membrane 300, a microfluidic board 400 with water-resisting property and absorbent paper 500. The upper cover 100 of the card shell is provided with a window 302 for embedding the detection membrane 300, the edge of the window 302 and the edge of the upper cover 100 of the card shell form a slope, the window 302 is recessed downwards, the detection membrane 300 is fixedly connected to the window 302 by four small convex columns 301 at the bottom of the window 302, the window 302 is arranged at the upper side of a microfluidic control plate 400, the microfluidic control plate 400 moves between the upper cover 100 of the card shell and the lower cover 200 of the card shell to enable the liquid permeable chip to be absorbed by the absorbent paper 500, the microfluidic control plate 400 and the upper cover 100 of the card shell can form a cavity for bearing liquid, meanwhile, the lower part of the microfluidic control plate 400 is made of absorbent material which can absorb the waste liquid after reaction, the chip carrier is a liquid permeable nitrocellulose membrane which is arranged above the microfluidic control plate 400 and fixedly supports a protein microarray, and the microfluidic control plate 400 can also be plastic, metal or kraft paper, the micro-fluidic plate 400 controls the reaction of the liquid and the protein loaded on the paper chip, a water absorption device is designed under the micro-fluidic plate 400, and after the reaction is finished, the micro-fluidic plate 400 is pulled, and the liquid permeation chip is absorbed by the water absorption paper 500.

The present invention is not limited to the above preferred embodiments, and any modifications, equivalent substitutions and improvements made within the spirit and principle of the present invention should be included in the protection scope of the present invention.