Four-item fan-shaped detection disc for thrombus

Abstract

The utility model discloses a thrombus four-item fan-shaped detection disc which comprises a disc body with a fan-shaped structure, and a sample adding groove, a separation groove connected with the sample adding groove and a quantitative unit connected with the separation groove which are sequentially arranged on the disc body from the rotating center of the disc body to the peripheral direction, the four detection units are connected with the quantitative unit, the at least one second waste liquid tank is respectively connected with the four detection units, a group of freeze-drying ball combinations for detecting four markers of thrombus are respectively preset in the four detection units, and each group of freeze-drying ball combinations comprises antibody-coated magnetic bead freeze-drying balls and enzyme-labeled antibody freeze-drying balls; the magnetic bead freeze-drying ball is pre-arranged in the detection groove, and the enzyme-labeled antibody freeze-drying ball is pre-arranged in the first injection groove. The detection disc is matched with detection equipment to realize the functions of whole blood separation, plasma quantification, freeze-drying dissolution, reagent mixing, rotary incubation, centrifugal cleaning and the like, four set results of thrombus are obtained through single whole blood, the detection sensitivity is high, the results are accurate and reliable, and the repeatability is good.

Description

Technical Field

[0001] The utility model relates to the technical field of biochemical detection, in particular to a four-item fan-shaped detection disc for thrombosis detection. Background Art

[0002] Thrombosis is a disease with high mortality that has a hidden onset and sudden attack. Whether it is arterial thrombosis, venous thrombosis or microvascular thrombosis, it poses a great threat to human health. A number of studies have shown that early detection and diagnosis of high-risk thrombosis populations in various clinical departments can achieve early intervention and early treatment, effectively reducing the harm of thrombosis to human health. Existing research has shown that when the body is in a pre-thrombotic state, the vascular endothelium, coagulation and fibrinolysis systems have changed. TAT (thrombin-antithrombin III complex), PIC (plasmin-α2 plasmin inhibitor complex), TM (thrombomodulin) and tPAI·C (tissue-type plasminogen activator-plasminogen activator inhibitor-1 complex) are effective indicators reflecting the early changes of the body's vascular endothelium, coagulation and fibrinolysis systems, and are applicable to early diagnosis, risk assessment and treatment efficacy evaluation of thrombosis in high-risk populations in various clinical disciplines and thrombosis risk screening in healthy populations.

[0003] At present, chemiluminescence immunoassay has been widely used in the detection of four items of thrombosis due to its simple operation, strong specificity, high sensitivity and short detection time. In chemiluminescence immunoassay, magnetic particle immunoassay technology is used to synthesize magnetic solid-phase particles of a certain particle size using high molecular materials as carriers, and various immunoreactive substances such as antibodies or antigens with specific affinity are coated by physical adsorption, chemical coupling and other methods. It has the characteristics of fast separation speed, high efficiency, good repeatability, simple operation, and does not affect the biological properties and functions of the separated cells or other biological materials. It can move directionally under the action of an external magnetic field, enabling the separation, concentration or purification of certain special components; however, at present, chemiluminescence immunoassay generally uses liquid reagents for detection, and due to the need for refrigerated storage of liquid reagents, the transportation and storage costs are relatively high.

[0004] With the development of detection technology, microfluidic chip technology has been widely used in many fields due to its advantages of high throughput, integration, portability, easy operation and low cost. How to combine microfluidic chip technology with chemiluminescence immunoassay to more quickly achieve the detection of four items of thrombosis is an urgent technical problem in this field. Summary of the Invention

[0005] In order to overcome the deficiencies of the prior art, the purpose of the present utility model is to provide a four-item thrombosis fan-shaped detection disc, which can realize functions such as whole blood separation, plasma quantification, lyophilized dissolution, reagent mixing, rotary incubation, centrifugal cleaning, etc. in cooperation with detection equipment. By using a single whole blood sample, the results of the four-item thrombosis package can be obtained, with high detection sensitivity, accurate and reliable results, and good repeatability.

[0006] To solve the above problems, the following solutions are adopted in the present utility model:

[0007] The present utility model includes a four-item thrombosis fan-shaped detection disc, which comprises a disc body with a fan-shaped structure, and a sample addition groove arranged on the disc body in sequence from the rotation center of the disc body to the peripheral direction, a separation groove connected to the sample addition groove, a quantification unit connected to the separation groove, four detection units connected to the quantification unit, and at least one second waste liquid groove respectively connected to the four detection units. The detection unit includes a combination of a detection groove and an injection groove. The detection groove is connected to the second waste liquid groove. The injection groove combination includes a first injection groove and a second injection groove respectively connected to the detection groove. The injection groove combination is located between the quantification unit and the detection groove. A set of lyophilized ball combinations for detecting four thrombosis markers are respectively preset in the four detection units. Each set of lyophilized ball combinations includes magnetic bead lyophilized balls coated with antibodies and enzyme-labeled antibody lyophilized balls. The magnetic bead lyophilized balls are preset in the detection groove, and the enzyme-labeled antibody lyophilized balls are preset in the first injection groove.

[0008] Preferably, the lyophilized ball combinations are respectively TAT lyophilized ball combination, PIC lyophilized ball combination, TM lyophilized ball combination, and tPAI·C lyophilized ball combination.

[0009] Preferably, a limiting sill for restricting the movement of the enzyme-labeled antibody lyophilized balls is arranged in the first injection groove.

[0010] Preferably, a connection end connected to the third microchannel is arranged at the top of the second waste liquid groove, and diversion grooves are respectively arranged on both sides of it. The distance from the top end of the diversion groove to the rotation center of the disc body is less than the distance from the connection end to the rotation center of the disc body.

[0011] Preferably, a first accommodation cavity and a second accommodation cavity are sequentially arranged in the second waste liquid groove along the rotation center of the disc body to the peripheral direction. The depth of the first accommodation cavity is greater than the depth of the second accommodation cavity.

[0012] Compared with the prior art, the beneficial effects of the present utility model are as follows:

[0013] In the present utility model, the detection disk in cooperation with the detection equipment can realize functions such as whole blood separation, plasma quantification, freeze-drying dissolution, reagent mixing, rotary incubation, centrifugal cleaning, etc. The thrombus four-item package results can be obtained from a single whole blood sample, with high detection sensitivity, accurate and reliable results, and good repeatability. By pre-packaging the freeze-dried balls in combination in the detection disk, the transportation and consumable costs can be effectively reduced. The detection disk adopts a fan-shaped structure, and a circular disk is formed by combining multiple disk pieces, enabling the detection of multiple detection objects simultaneously, greatly improving the utilization rate of the disk pieces and reducing the waste of disk pieces. By combining the detection disk with the freeze-dried ball reagent, the attributes of the detection disk and the reagent as consumables and main raw materials respectively can be changed. Placing the two products together can form a composite consumable, improving the convenience for users during use. At the same time, the package disk can be freely and specifically configured according to needs, greatly meeting the customized requirements of customers. BRIEF DESCRIPTION OF THE DRAWINGS

[0014] Figure 1 It is a schematic structural diagram of the thrombus four-item fan-shaped detection disk in a preferred embodiment of the present utility model;

[0015] Figure 2 is Figure 1 a schematic structural diagram of the first injection groove in;

[0016] Figure 3 is a schematic structural diagram of the second waste liquid groove in a specific embodiment;

[0017] Among them, the reference numerals are explained as follows: 1 is the disk body, 10 is the positioning groove, 11 is the first positioning groove, 12 is the second positioning groove, 13 is the dial, 21 is the sample addition groove, 22 is the separation groove, 23 is the first microchannel, 241 is the shunt channel, 242 is the first waste liquid groove, 243 is the quantification groove, 244 is the first ventilation hole, 25 is the second microchannel, 261 is the detection groove, 262 is the first injection groove, 2621 is the limiting sill, 263 is the second injection groove, 27 is the third microchannel, 28 is the second waste liquid groove, 281 is the second ventilation hole, 282 is the connection end, 283 is the diversion groove, 284 is the first accommodation cavity, 285 is the second accommodation cavity, 30 is the antibody-coated magnetic bead freeze-dried ball, and 31 is the enzyme-labeled antibody freeze-dried ball. DETAILED DESCRIPTION OF THE EMBODIMENTS

[0018] The present utility model will be further described in detail below with reference to the accompanying drawings.

[0019] Referring to Figure 1, which is a schematic structural diagram of a four-item thrombus fan-shaped detection disc in a preferred embodiment of the present utility model. The four-item thrombus fan-shaped detection disc includes a disc body 1 with a fan-shaped structure, and a sample adding groove 21 for adding whole blood samples, a separation groove 22 connected to the sample adding groove 21 for separating the whole blood samples, a quantification unit connected to the separation groove 22 for quantitatively shunting the plasma, four detection units connected to the quantification unit, and at least one second waste liquid groove 28 for collecting waste liquid respectively connected to the four detection units. The detection unit includes a detection groove 261 for freeze-drying dissolution, reagent mixing, rotary incubation, centrifugal cleaning and detection, and an injection groove combination. The detection groove 261 is connected to the second waste liquid groove 28. The injection groove combination includes a first injection groove 262 and a second injection groove 263 respectively connected to the detection groove 261. The injection groove combination is located between the quantification unit and the detection groove 261. The separation groove 22 is connected to the quantification unit through a first microchannel 23. The whole blood samples are separated through the first microchannel 23, and the flow of plasma in the whole blood samples to the quantification unit is controlled. The quantification unit is connected to the detection groove 261 through a second microchannel 25. The flow of the quantified plasma into the detection groove 261 is controlled through the second microchannel 25. The detection groove 261 is connected to the second waste liquid groove 28 through a third microchannel 27. The flow of waste liquid into the second waste liquid groove 28 is controlled through the third microchannel 27. A freeze-dried ball combination for detecting four thrombus markers is pre-set in each of the four detection units. Each group of freeze-dried ball combinations includes a magnetic bead freeze-dried ball 30 coated with an antibody and an enzyme-labeled antibody freeze-dried ball 31. The magnetic bead freeze-dried ball 30 is pre-set in the detection groove 261, and the enzyme-labeled antibody freeze-dried ball 31 is pre-set in the first injection groove 262. The disc body adopts a fan-shaped structure, and a circular disc is formed by combining multiple disc pieces, which can simultaneously realize the detection of multiple detection objects, greatly improving the utilization rate of the disc pieces and reducing the waste of the disc pieces. The detection disc, in cooperation with the detection equipment, can realize functions such as whole blood separation, plasma quantification, freeze-drying dissolution, reagent mixing, rotary incubation, centrifugal cleaning, etc. The four-item thrombus package results can be obtained from a single whole blood sample. The detection sensitivity is high, the results are accurate and reliable, and the repeatability is good. By pre-packaging the freeze-dried ball combination in the detection disc, the transportation and consumable costs can be reduced.

[0020] Specifically, the freeze-dried ball combinations for detecting four thrombus markers used in this application are respectively a TAT freeze-dried ball combination (including a magnetic bead freeze-dried ball coated with TAT antibody and an alkaline phosphatase-labeled TAT antibody freeze-dried ball), a PIC freeze-dried ball combination (including a magnetic bead freeze-dried ball coated with PIC antibody and an alkaline phosphatase-labeled PIC antibody freeze-dried ball), a TM freeze-dried ball combination (including a magnetic bead freeze-dried ball coated with TM antibody and an alkaline phosphatase-labeled TM antibody freeze-dried ball), and a tPAI·C freeze-dried ball combination (including a magnetic bead freeze-dried ball coated with tPAI·C antibody and an alkaline phosphatase-labeled tPAI·C antibody freeze-dried ball).

[0021] A semi-permeable membrane can be provided in the first microchannel 23 above to separate the sample in the separation tank 22 during centrifugation, and the plasma enters the quantification unit through the first microchannel; a microfluidic valve can be provided in the second microchannel 25 to strengthen the control of the plasma flow direction during centrifugation; a microfluidic valve can be provided in the third microchannel 27 to strengthen the control of the waste liquid flow direction during centrifugation.

[0022] Specifically, the central angle of the disk body 1 is 90°, and a first positioning groove 11 and a second positioning groove 12 are provided on the outer periphery of the disk body 1. The sizes of the first positioning groove 11 and the second positioning groove 12 are different and are used for positioning during the installation of the disk. At this time, the positioning groove 10 is preferably set to be arc-shaped or U-shaped for easy installation and disassembly.

[0023] In another specific embodiment, a positioning groove 10 is provided at the center of the disk body 1. In a specific embodiment, the positioning groove 10 is selected to have a structure such as a W shape or a wavy shape for easy positioning during the installation of the disk.

[0024] Specifically, a dial 13 for facilitating the disassembly of the detection disk is provided on the outer periphery of the disk body 1.

[0025] Specifically, referring again to Figure 1 , the quantification unit includes a shunt channel 241 for shunting the plasma, a first waste liquid tank 242 for receiving the excess plasma, and four quantification tanks 243 for quantitatively measuring the plasma entering the detection tank 27. One end of the shunt channel 241 is connected to the separation tank 22 through the first microchannel 23, and the other end is connected to the first waste liquid tank 242. The four quantification tanks 243 are connected to the shunt channel 241, and the four quantification tanks 243 are connected to the detection tank 261 through the second microchannel 25. Further, the shunt channel 241 is generally arc-shaped, and the distance from the end close to the first microchannel 23 to the rotation center of the disk body 1 is not greater than the distance from the end close to the first waste liquid tank 242 to the rotation center of the disk body 1, which is convenient for the excess plasma to be collected into the first waste liquid tank 242.

[0026] Specifically, a first ventilation hole 244 is connected to the first waste liquid tank 242, and a second ventilation hole 281 is connected to the second waste liquid tank 28. The ventilation hole structure can adjust the air pressure in the disk body 1, facilitate the flow of plasma or waste liquid in the disk body 1, and facilitate their collection.

[0027] Specifically, referring to Figure 2 , a limiting sill 2621 for restricting the movement of the enzyme-labeled antibody lyophilized ball 31 is provided in the first injection tank 262. The limiting sill 2621 and the first injection tank 262 cooperate to form a receiving space for the enzyme-labeled antibody lyophilized ball 31, and a certain fixation is formed on the position of the enzyme-labeled antibody lyophilized ball 31.

[0028] Specifically, referring to Figure 3, in a specific embodiment, a connection end 282 connected to the third microchannel 27 is provided at the top of the second waste liquid tank 28. Flow guiding grooves 283 are respectively provided on both sides of the connection end 282. The distance from the top end of the flow guiding groove 283 to the rotation center of the disc body 1 is less than the distance from the connection end 282 to the rotation center of the disc body 1. The flow guiding grooves 283 can guide the solution in the second waste liquid tank 28 during forward and reverse rotation and mixing, avoiding the backflow of the solution into the third microchannel 27.

[0029] Refer again to Figure 3 , a first accommodation cavity 284 and a second accommodation cavity 285 are sequentially arranged in the second waste liquid tank 28 along the direction from the rotation center of the disc body 1 to the outer periphery. The depth of the first accommodation cavity 284 is greater than that of the second accommodation cavity 285. The second accommodation cavity 284 has a greater surface tension on the solution in the second waste liquid tank 28 compared to the first accommodation cavity 283, which can better ensure that the solution stays in the second accommodation cavity 284 and prevent the solution in the second waste liquid tank 28 from flowing back into the third microchannel 27 during forward and reverse rotation and mixing. At this time, in order to better dissolve the enzyme-labeled antibody freeze-dried beads 31, the enzyme-labeled antibody freeze-dried beads 31 are pre-placed in the second accommodation cavity 284.

[0030] Next, Figure 1 the usage principle of the thrombus four-item fan-shaped detection disc in

[0031] The first step is to add a whole blood sample into the sample addition groove of the detection disc;

[0032] The second step is that through centrifugation, the whole blood sample enters the separation groove, and the plasma in the whole blood sample in the separation groove is separated into the shunt flow channel through the first microchannel;

[0033] The third step is to continue centrifugation. After the plasma fills the quantitative groove, the excess plasma enters the first waste liquid tank;

[0034] The fourth step is to adjust the centrifugation speed so that the plasma in the quantitative groove breaks through the second microchannel and enters the detection groove to preliminarily dissolve the antibody-coated magnetic bead freeze-dried beads in the detection groove;

[0035] Step 5: Stop centrifugation, add the activation solution into the second injection tank, centrifuge again to allow the activation solution to enter the detection tank and completely dissolve the freeze-dried ball combination. Then, continuously change the centrifugation direction, rotate in both forward and reverse directions, and auxiliary treatments such as heating the detection tank can be carried out to mix and incubate the plasma with the antibody-coated magnetic beads, obtaining a mixed solution containing magnetic bead-antibody-antigen intermediates. Adjust the centrifugation speed to allow the waste liquid in the mixed solution in the detection tank to break through the third microchannel and enter the second waste liquid tank. At the same time, with the help of an external magnetic field, the magnetic bead-antibody-antigen intermediates are retained in the detection tank. Stop centrifugation, add the cleaning solution into the second injection tank, centrifuge again to allow the cleaning solution to enter the detection tank, and then continuously change the centrifugation direction, rotate in both forward and reverse directions to thoroughly clean the magnetic bead-antibody-antigen intermediates with the cleaning solution. Adjust the centrifugation speed again to allow the cleaning solution containing impurities to break through the third microchannel and enter the second waste liquid tank. At the same time, with the help of an external magnetic field, the highly purified magnetic bead-antibody-antigen intermediates after cleaning are retained in the detection tank.

[0036] Step 6: Stop centrifugation, add the activation solution into the first injection tank to completely dissolve the freeze-dried enzyme-labeled antibody balls. Centrifuge again to allow the activated enzyme-labeled antibody solution to enter the detection tank, and then continuously change the centrifugation direction, rotate in both forward and reverse directions, and auxiliary treatments such as heating the detection tank can be carried out to mix and incubate the magnetic bead-antibody-antigen intermediates and the enzyme-labeled antibody, obtaining a mixed solution containing antigen-antibody sandwich conjugates.

[0037] Step 7: Adjust the centrifugation speed to allow the waste liquid in the mixed solution in the detection tank to break through the third microchannel and enter the second waste liquid tank. At the same time, with the help of an external magnetic field, the antigen-antibody sandwich conjugates are retained in the detection tank. Stop centrifugation, add the cleaning solution into the injection tank, centrifuge again to allow the cleaning solution to enter the detection tank, and then continuously change the centrifugation direction, rotate in both forward and reverse directions to thoroughly clean the antigen-antibody sandwich conjugates with the cleaning solution. Adjust the centrifugation speed again to allow the cleaning solution containing impurities to break through the third microchannel and enter the second waste liquid tank. At the same time, with the help of an external magnetic field, the highly purified antigen-antibody sandwich conjugates after cleaning are retained in the detection tank.

[0038] Step 8: Stop centrifugation, add the luminescent substrate into the second injection tank, centrifuge again to allow the luminescent substrate to enter the detection tank, rotate in both forward and reverse directions, and auxiliary treatments such as heating the detection tank can be carried out to mix and react the antigen-antibody sandwich conjugates with the luminescent substrate. After the reaction ends, the luminescence values in the four detection tanks can be detected and analyzed simultaneously by a detection device.

[0039] The embodiments of this specific implementation manner are all preferred embodiments of the present invention, and do not limit the protection scope of the present invention accordingly. Therefore, all equivalent changes made according to the structure, shape, and principle of the present invention should be covered within the protection scope of the present invention.

Claims

1. A four-item sector-shaped thrombus detection disc, characterized in that: The invention comprises a disc body with a fan-shaped structure, and a sample loading slot, a separation slot connected to the sample loading slot, a certain amount unit connected to the separation slot, four detection units connected to the quantitative unit, and at least one second waste liquid slot respectively connected to the four detection units, the detection unit comprises a detection slot and an injection slot combination, the detection slot is connected to the second waste liquid slot, the injection slot combination comprises a first injection slot and a second injection slot respectively connected to the detection slot, and the injection slot combination is located between the quantitative unit and the detection slot; a group of freeze-dried ball combinations for detecting four markers of thrombosis are respectively preset in the four detection units, and each group of freeze-dried ball combinations comprises antibody-coated magnetic bead freeze-dried balls and enzyme-labeled antibody freeze-dried balls; The magnetic bead freeze-dried ball is preset in the detection tank, and the enzyme-labeled antibody freeze-dried ball is preset in the first injection tank.

2. The four-item sector-shaped thrombus detection disc according to claim 1, characterized in that: The freeze-dried ball combinations are respectively TAT freeze-dried ball combination, PIC freeze-dried ball combination, TM freeze-dried ball combination and tPAI·C freeze-dried ball combination.

3. The four-item sector-shaped thrombus detection disc according to claim 1, characterized in that: A limit threshold for limiting the movement of enzyme-labeled antibody freeze-dried beads is arranged in the first injection groove.

4. The four-item sector-shaped thrombus detection disc according to claim 1, characterized in that: The top of the second waste liquid tank is provided with a connection end connected to the third microchannel, and two sides thereof are provided with guide grooves, and the distance between the top of the guide groove and the rotation center of the disk body is smaller than the distance between the connection end and the rotation center of the disk body.

5. The four-item sector-shaped thrombus detection disc according to claim 1, characterized in that: A first accommodating cavity and a second accommodating cavity are sequentially arranged in the second waste liquid tank along the rotation center of the disk body toward the outer circumference, and the depth of the first accommodating cavity is greater than the depth of the second accommodating cavity.

Images