Quadruple fan-shaped detection disc for chest pain

Abstract

The invention discloses a chest pain quadruple 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 grooves are connected to the quantifying unit; the at least one second waste liquid groove is connected with the four detection grooves respectively; the at least one injection groove is positioned between the quantitative unit and the detection groove and is connected to the detection groove; a group of freeze-drying ball combination for detecting chest pain markers is preset in each of the four detection grooves, and each group of freeze-drying ball combination comprises antibody-coated magnetic bead freeze-drying balls and enzyme-labeled antibody freeze-drying balls. 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, a chest pain quadruple package result is obtained through single whole blood, the detection sensitivity is high, the result is accurate and reliable, and the repeatability is good.

Description

Technical Field

[0001] This invention relates to the field of biochemical detection technology, and in particular to a four-piece fan-shaped detection disc for chest pain. Background Technology

[0002] Acute coronary events, acute pulmonary embolism, acute aortic dissection, and chest wall masses are the main non-traumatic emergencies besides stroke, collectively known as the tetrad of chest pain. Because these four conditions all manifest as chest pain, with no specificity between symptoms and a sudden onset, failure to diagnose them quickly in clinical practice is extremely dangerous. Commonly used biomarkers for diagnosing chest pain include cardiac troponin I (cTnI), B-type natriuretic peptide (BNP), D-dimer (D-Dimer), myoglobin (MYO), creatine kinase isoenzyme (CK-MB), heart-type fatty acid-binding protein (H-FABP), myosin-binding protein C (MYBPC), and peptides.

[0003] Currently, the main methods for detecting chest pain using the quadruple assay include radioimmunoassay, chemiluminescence immunoassay, and colloidal gold colorimetry. Radioimmunoassay is time-consuming and its use is limited by the risk of radioactive contamination. Colloidal gold colorimetry is rapid, simple, and easy to observe, making it a fast semi-quantitative method. Chemiluminescence immunoassay is simple to operate, highly specific, sensitive, and has a short detection time, and is currently widely used. In chemiluminescence immunoassay, magnetic microparticle immunoassay technology utilizes polymer materials to synthesize magnetic solid-phase microparticles of a specific size as carriers. These microparticles are coated with various immunologically active substances, such as antibodies or antigens, with specific affinity through physical adsorption and chemical coupling. This method features fast separation speed, high efficiency, good reproducibility, simple operation, and no impact on the biological properties and functions of the separated cells or other biological materials. Under the influence of an external magnetic field, these microparticles can move directionally, allowing for the separation, concentration, or purification of certain specific components. However, currently, chemiluminescence immunoassay generally uses liquid reagents for detection, which require refrigeration, resulting in higher transportation and storage costs.

[0004] With the development of detection technology, microfluidic chip technology has been widely used in many fields due to its advantages such as high throughput, integration, convenience, ease of operation, and low cost. How to combine microfluidic chip technology with chemiluminescence immunoassay to achieve faster detection of four types of chest pain is a pressing technical problem that needs to be solved in this field. Summary of the Invention

[0005] To overcome the shortcomings of the existing technology, the purpose of this invention is to provide a chest pain quadruple sector-shaped detection disk, which, when used with detection equipment, can realize functions such as whole blood separation, plasma quantification, lyophilization and dissolution, reagent mixing, rotation incubation, and centrifugation washing. It can obtain chest pain quadruple package results from a single whole blood sample, with high detection sensitivity, accurate and reliable results, and good repeatability.

[0006] To solve the above problems, the solution adopted by the present invention is as follows: This invention includes a four-piece fan-shaped detection disc for chest pain, comprising a fan-shaped disc body, a sample loading groove arranged sequentially on the disc body from the center of rotation outwards, a separation groove connected to the sample loading groove, a quantitative unit connected to the separation groove, four detection grooves connected to the quantitative unit, and at least one second waste liquid groove connected to each of the four detection grooves; and at least one injection groove located between the quantitative unit and the detection grooves and connected to the detection grooves; each of the four detection grooves contains a set of lyophilized beads for detecting chest pain markers, each set of lyophilized beads including antibody-coated magnetic bead lyophilized beads and enzyme-labeled antibody lyophilized beads.

[0007] Preferably, the freeze-dried pellet combinations are cTnI freeze-dried pellet combination, MYO freeze-dried pellet combination, BNP freeze-dried pellet combination, and D-Dimer freeze-dried pellet combination.

[0008] Preferably, the separation tank is connected to the quantitative unit via a first microchannel, the quantitative unit is connected to the detection tank via a second microchannel, and the four detection tanks are respectively connected to the second waste liquid tank via a third microchannel.

[0009] Preferably, the quantitative unit includes a diversion channel, a first waste liquid tank and four quantitative tanks. One end of the diversion channel is connected to the separation tank through a first microchannel, and the other end is connected to the first waste liquid tank. The four quantitative tanks are connected to the diversion channel, and the four quantitative tanks are respectively connected to the detection tank through a second microchannel.

[0010] Preferably, the top of the detection groove is provided with a connecting end that connects to the second microchannel, and guide grooves are provided on both sides of the guide groove. The distance from the top of the guide groove to the rotation center of the disk is less than the distance from the connecting end to the rotation center of the disk.

[0011] Preferably, a first receiving cavity and a second receiving cavity are sequentially arranged in the detection groove along the rotation center of the disk towards the outer periphery, and the depth of the first receiving cavity is greater than the depth of the second receiving cavity.

[0012] Preferably, a positioning groove is provided at the center of the disc body, and a first positioning groove is provided on its outer periphery.

[0013] Preferably, the central angle of the disk is 90°.

[0014] Preferably, a second positioning groove is also provided on the outer periphery, and the first positioning groove and the second positioning groove have different dimensions.

[0015] Compared with the prior art, the beneficial effects of the present invention are as follows: In this invention, the detection disc, in conjunction with the detection equipment, can achieve functions such as whole blood separation, plasma quantification, lyophilization and dissolution, reagent mixing, rotary incubation, and centrifugal washing. It obtains a four-in-one chest pain test result from a single whole blood sample, exhibiting high sensitivity, accurate and reliable results, and good repeatability. By pre-packaging the lyophilized bulbs in the detection disc, transportation and consumable costs are reduced. The detection disc adopts a fan-shaped structure, combining multiple discs into a circular disc, allowing for simultaneous detection of multiple subjects, significantly improving disc utilization and reducing waste. By combining the detection disc with the lyophilized bulb reagent, the properties of the detection disc and reagent as consumables and main raw materials, respectively, are changed. Combining the two products creates a composite consumable, improving user convenience. Furthermore, the package discs can be customized to meet specific customer needs. Attached Figure Description

[0016] Figure 1 This is a schematic diagram of the structure of the chest pain quadruple sector detection disc in a preferred embodiment of the present invention; Figure 2 for Figure 1 Schematic diagram of the structure of the detection tank; The following are the annotations in the attached drawings: 1 is the disc body, 10 is the positioning groove, 11 is the first positioning groove, 12 is the second positioning groove, 13 is the lever, 21 is the sample loading groove, 22 is the separation groove, 23 is the first microchannel, 241 is the diversion channel, 242 is the first waste liquid tank, 243 is the quantitative tank, 244 is the first vent, 25 is the second microchannel, 26 is the injection tank, 27 is the detection tank, 271 is the connection end, 272 is the guide groove, 273 is the first accommodating cavity, 274 is the second accommodating cavity, 28 is the third microchannel, 29 is the second waste liquid tank, 291 is the second vent, 30 is the antibody-coated magnetic bead lyophilized bulb, and 31 is the enzyme-labeled antibody lyophilized bulb. Detailed Implementation

[0017] The present invention will be further described in detail below with reference to the accompanying drawings.

[0018] Reference Figure 1This is a schematic diagram of a quadruple sector-shaped chest pain detection disk according to a preferred embodiment of the present invention. The quadruple sector-shaped chest pain detection disk includes a sector-shaped disk body 1, and a sample addition slot 21 for adding whole blood samples, a separation slot 22 for separating whole blood samples, a quantitative unit for quantitatively separating plasma, four detection slots 27 for lyophilization dissolution, reagent mixing, rotational incubation, centrifugal washing, and detection, and at least one second waste liquid tank for collecting waste liquid, arranged sequentially along the rotation center of the disk body 1 outwards. 29; The sample loading tank 21 is connected to the separation tank 22. The separation tank 22 is connected to the quantitative unit through the first microfluidic channel 23, which separates the whole blood sample and controls the flow of plasma from the whole blood sample to the quantitative unit. The quantitative unit is connected to the detection tank 27 through the second microfluidic channel 25, which controls the flow of a quantified amount of plasma into the detection tank 27. The detection tank 27 is connected to the second waste liquid tank 29 through the third microfluidic channel 28, which controls the flow of waste liquid into the second waste liquid tank 29. Each of the four detection tanks contains a set of lyophilized bulbs for detecting chest pain markers. Each set of lyophilized bulbs includes antibody-coated magnetic bead lyophilized bulbs 30 and enzyme-labeled antibody lyophilized bulbs 31. The disc body adopts a fan-shaped structure, and multiple discs are combined into a circular disc, which can simultaneously detect multiple objects, greatly improving the utilization rate of the discs and reducing the waste of discs. The detection disc, together with the detection equipment, can realize functions such as whole blood separation, plasma quantification, lyophilization and dissolution, reagent mixing, rotation incubation, centrifugation and washing. It can obtain the results of a four-in-one chest pain test 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 lyophilized bulbs in the detection disc, the cost of transportation and consumables is reduced.

[0019] Specifically, this application uses the following lyophilized bead combinations for detecting chest pain biomarkers: a cTnI lyophilized bead combination (including cTnI antibody-coated magnetic bead lyophilized beads and alkaline phosphatase-labeled cTnI antibody lyophilized beads), a MYO lyophilized bead combination (including MYO antibody-coated magnetic bead lyophilized beads and alkaline phosphatase-labeled MYO antibody lyophilized beads), a BNP lyophilized bead combination (including BNP antibody-coated magnetic bead lyophilized beads and alkaline phosphatase-labeled BNP antibody lyophilized beads), and a D-Dimer lyophilized bead combination (including D-Dimer antibody-coated magnetic bead lyophilized beads and alkaline phosphatase-labeled D-Dimer antibody lyophilized beads). It should be noted that all the aforementioned lyophilized beads were prepared using conventional methods in the art, and will not be described in detail here.

[0020] A semi-permeable membrane can be installed in the first microchannel 23 to separate the sample in the separation tank 22 during centrifugation, and the plasma enters the quantitative unit through the first microchannel; a microfluidic valve can be installed in the second microchannel 25 to enhance the control of the plasma flow direction during centrifugation; a microfluidic valve can be installed in the third microchannel 28 to enhance the control of the waste liquid flow direction during centrifugation.

[0021] 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 first positioning groove 11 and the second positioning groove 12 have different sizes and are used for positioning during disk installation. In this case, the positioning groove 10 is preferably set to an arc shape or a U shape to facilitate installation and disassembly.

[0022] In another specific embodiment, a positioning groove 10 is provided at the center of the disk body 1. In one specific embodiment, the positioning groove 10 is selected from a W-shaped or wave-shaped structure to facilitate positioning during disk installation.

[0023] Specifically, a paddle 13 is provided on the outer periphery of the disc body 1 to facilitate the disassembly of the testing disc.

[0024] For details, please refer to again. Figure 1 The quantitative unit includes a diversion channel 241 for diverting plasma, a first waste liquid tank 242 for receiving excess plasma, and four quantitative tanks 243 for quantitatively measuring plasma entering the detection tank 27. One end of the diversion channel 241 is connected to the separation tank 22 via a first microchannel 23, and the other end is connected to the first waste liquid tank 242. The four quantitative tanks 243 are connected to the diversion channel 241 and are connected to the detection tank 27 via a second microchannel 25. Furthermore, the diversion channel 241 is approximately arc-shaped, and the distance from the end near the first microchannel 23 to the rotation center of the disk 1 is no greater than the distance from the end near the first waste liquid tank 242 to the rotation center of the disk 1, facilitating the collection of excess plasma into the first waste liquid tank 242.

[0025] Specifically, the first waste liquid tank 242 is connected to a first vent hole 244, and the second waste liquid tank 29 is connected to a second vent hole 291. The vent hole structure can adjust the air pressure inside the plate 1, which facilitates the flow of plasma or waste liquid inside the plate 1 and makes it easier for them to be collected.

[0026] Specifically, refer to Figure 2 The top of the detection tank 27 is provided with a connecting end 271 that connects to the second microchannel 25. Guide channels 272 are respectively provided on both sides of the connecting end 271. The distance from the top of the guide channel 272 to the rotation center of the disk 1 is less than the distance from the connecting end 271 to the rotation center of the disk 1. The guide channels 272 can guide the solution in the detection tank 27 during forward and reverse rotation mixing, preventing the solution from flowing back into the second microchannel 25.

[0027] Refer again Figure 2In one specific embodiment, a first receiving cavity 273 and a second receiving cavity 274 are sequentially arranged along the outer periphery of the rotation center of the disk 1 within the detection tank 27. The depth of the first receiving cavity 273 is greater than the depth of the second receiving cavity 274. The second receiving cavity 274 has a greater surface tension on the solution in the detection tank 27 compared to the first receiving cavity 273, which can better ensure that the solution remains in the second receiving cavity 274 and prevent the solution in the detection tank 27 from flowing back into the second microchannel 25 when mixed by forward and reverse rotation. At this time, in order to ensure better dissolution of the freeze-dried ball assembly, the freeze-dried ball assembly is pre-placed in the second receiving cavity 274.

[0028] The following is about Figure 1 The working principle of the quadruple sector-shaped detection disc for chest pain is further explained below: The first step is to add a whole blood sample to the sample compartment of the testing tray; The second step involves centrifugation, during which the whole blood sample enters the separation tank, and the plasma in the whole blood sample is separated into the diversion channel via the first microfluidic channel. The third step is to continue centrifugation. After the plasma fills the quantitative tank, the excess plasma enters the first waste liquid tank. The fourth step is to adjust the centrifugation speed so that the plasma in the quantitative tank breaks through the second microchannel and enters the detection tank to initially dissolve the lyophilized pellets in the detection tank. Fifth step: Stop centrifugation, add activation solution to the injection tank, centrifuge again to allow the activation solution to enter the detection tank and completely dissolve the lyophilized beads; then continuously change the centrifugation direction, by rotating in both directions, and may also assist in heating the detection tank, to achieve the mixing and incubation of plasma with antibody-coated magnetic beads and enzyme-labeled antibodies, and obtain a mixture containing antigen-antibody sandwich conjugates. Step 6: Adjust the centrifugation speed to allow the waste liquid in the mixture 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 conjugate is retained in the detection tank. Stop centrifugation, add cleaning solution to the injection tank, and centrifuge again to allow the cleaning solution to enter the detection tank. Then, continuously change the centrifugation direction and achieve thorough cleaning of the antigen-antibody sandwich conjugate by rotating in both directions. 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 cleaned high-purity antigen-antibody sandwich conjugate is retained in the detection tank. Step 7: Stop centrifugation, add the luminescent substrate to the injection tank, centrifuge again to allow the luminescent substrate to enter the detection tank, and achieve the mixing and reaction of the antigen-antibody sandwich conjugate and the luminescent substrate by rotating in both directions and by heating the detection tank. After the reaction is completed, the luminescence values ​​in the four detection tanks can be detected and analyzed simultaneously by the detection equipment.

[0029] The embodiments described herein are preferred embodiments of the present invention and are not intended to limit the scope of protection of the present invention. Therefore, all equivalent changes made in accordance with the structure, shape, and principle of the present invention should be covered within the scope of protection of the present invention.

Claims

1. A four-piece sector-shaped chest pain detection disc, characterized in that, The device includes a fan-shaped disk body, a sample loading groove arranged sequentially on the disk body from the rotation center outwards, a separation groove connected to the sample loading groove, a quantitative unit connected to the separation groove, four detection grooves connected to the quantitative unit, and at least one second waste liquid groove connected to the four detection grooves respectively. And at least one injection tank located between the quantitative unit and the detection tank, connected to the detection tank; Each of the four detection chambers contains a set of lyophilized beads for detecting chest pain markers. Each set of lyophilized beads includes antibody-coated magnetic bead lyophilized beads and enzyme-labeled antibody lyophilized beads.

2. The chest pain quadruple sector-shaped detection disc as described in claim 1, characterized in that, The freeze-dried pellet combinations are cTnI freeze-dried pellet combination, MYO freeze-dried pellet combination, BNP freeze-dried pellet combination and D-Dimer freeze-dried pellet combination.

3. The chest pain quadruple sector-shaped detection disc as described in claim 1, characterized in that, The separation tank is connected to the quantitative unit through the first microchannel, the quantitative unit is connected to the detection tank through the second microchannel, and the four detection tanks are respectively connected to the second waste liquid tank through the third microchannel.

4. The chest pain quadruple sector-shaped detection disc as described in claim 3, characterized in that, The quantitative unit includes a diversion channel, a first waste liquid tank, and four quantitative tanks. One end of the diversion channel is connected to the separation tank through a first microchannel, and the other end is connected to the first waste liquid tank. The four quantitative tanks are connected to the diversion channel, and the four quantitative tanks are respectively connected to the detection tank through a second microchannel.

5. The chest pain quadruple sector-shaped detection disc as described in claim 3, characterized in that, The top of the detection groove is provided with a connection end that connects to the second microchannel, and guide grooves are provided on both sides of it. The distance from the top of the guide groove to the center of rotation of the disk is less than the distance from the connection end to the center of rotation of the disk.

6. The chest pain quadruple sector-shaped detection disc as described in claim 3, characterized in that, The detection groove contains a first accommodating cavity and a second accommodating cavity arranged sequentially from the rotation center of the disc outwards. The depth of the first accommodating cavity is greater than the depth of the second accommodating cavity.

7. The chest pain quadruple sector-shaped detection disc as described in any one of claims 1-6, characterized in that, A positioning groove is provided at the center of the disc body, and a first positioning groove is provided on its outer periphery.

8. The chest pain quadruple sector-shaped detection disc as described in claim 7, characterized in that, The central angle of the disk is 90°.

9. The chest pain quadruple sector-shaped detection disc as described in claim 7, characterized in that, A second positioning groove is also provided on the outer periphery, and the first positioning groove and the second positioning groove have different dimensions.

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