A reaction disc for rapid detection of c-reactive protein

Abstract

The utility model discloses a reaction disc for rapid detection of C reaction protein belongs to C reaction protein detection technical field, including disc, be equipped with reaction subassembly on the disc, the reaction subassembly contains first liquid storage bin, second liquid storage bin and reaction bin, be equipped with multiple groups micro -flow channel on the disc, first liquid storage bin and second liquid storage bin are linked with reaction bin through corresponding micro -flow channel respectively, through the drive disc rotation, need not the complex mechanical device such as external pump, valve, through the adjustment disc rotation speed, can accurate control solution's flow (such as starting, stopping, switching path) in micro -flow channel, promote sample and solution reagent's uniform mixing, and mixed efficiency compared with passive diffusion has the remarkable improvement, and reaction is sufficient.

Description

Technical Field

[0001] This utility model relates to the field of C-reactive protein detection technology, and in particular to a reaction disk for rapid C-reactive protein detection. Background Technology

[0002] C-reactive protein (CRP), as a key inflammatory marker, has irreplaceable and multifaceted value in clinical diagnosis and treatment, and its detection and application span multiple disciplines in disease management. For example, in the field of infectious diseases, dynamic changes in CRP levels are an important basis for distinguishing between bacterial and viral infections. Furthermore, by monitoring the decline in CRP levels during treatment, the effectiveness of anti-infective therapy can be quickly assessed, guiding adjustments to the treatment course and shortening the average hospital stay by 1-2 days. In addition, CRP plays a crucial role in connective tissue diseases (such as monitoring active rheumatoid arthritis), metabolic syndrome (positively correlated with insulin resistance), cancer (an auxiliary indicator of tumor progression and metastasis), neurological diseases (assessment of inflammatory responses after cerebral hemorrhage), and in obstetrics and gynecology, pediatrics, and surgery (early warning of postoperative infection).

[0003] Therefore, CRP testing is crucial. Traditional CRP testing relies on large biochemical analyzers, which takes a long time from sample collection to report generation and requires professional laboratory support, making it difficult to meet the immediate needs of emergency departments, primary healthcare institutions, and POCT scenarios.

[0004] In contrast, the latest microfluidic detection, by constructing channels and reaction chambers in a (10-100) μm microscale space, shortens the diffusion distance of biomolecules to 1 / 100 of that in traditional systems. The probability of antigen-antibody collision increases exponentially, the kinetic rate is greatly improved, and the immune reaction that originally took several hours can be completed in just a few minutes.

[0005] Microfluidic detection devices for C-reactive protein detection have been developed. The core of these devices is to drop various solution reagents onto a reaction plate for reaction. However, they often require external pumps or modules to control the flow of the solution and accelerate the immune reaction. This process is cumbersome and has low reaction efficiency. There is still much room for improvement in terms of sample processing convenience and the versatility of detection and analysis. Utility Model Content

[0006] The purpose of this invention is to solve the technical problems in the prior art, which require external pumps or modules to control solution flow and accelerate immune reactions, resulting in cumbersome operation and low reaction efficiency. Therefore, this invention proposes a reaction disk for rapid detection of C-reactive protein.

[0007] To achieve the above objectives, the present invention adopts the following technical solution:

[0008] A reaction disk for rapid detection of C-reactive protein includes a disk on which a reaction assembly is provided; the reaction assembly includes a first liquid reservoir, a second liquid reservoir, and a reaction chamber; the disk is provided with multiple sets of microfluidic channels; the first liquid reservoir and the second liquid reservoir are respectively connected to the reaction chamber through corresponding microfluidic channels.

[0009] Preferably, the reaction assembly further includes a third liquid storage chamber; the third liquid storage chamber is also connected to the reaction chamber through a corresponding microfluidic channel.

[0010] Preferably, the reaction assembly further includes a centrifuge chamber; the centrifuge chamber is also connected to the reaction chamber via a corresponding microfluidic channel.

[0011] Preferably, the width of the microfluidic channel is 0.6-1 mm.

[0012] Preferably, the connections between the microfluidic channel and the first, second, and third liquid storage tanks are all chamfered.

[0013] Preferably, the number of reaction components is 2-5.

[0014] Preferably, the number of reaction components is three.

[0015] Preferably, the disc is further provided with a heating chamber; the second liquid storage chamber is located inside the heating chamber.

[0016] Compared with the prior art, this utility model provides a reaction disk for rapid detection of C-reactive protein, which has the following beneficial effects:

[0017] 1. This rapid detection reaction disk for C-reactive protein, by driving the disk to rotate, eliminates the need for complex mechanical devices such as external pumps and valves. By adjusting the disk rotation speed, the flow of solution in the microchannel can be precisely controlled (such as starting, stopping, and switching paths), promoting uniform mixing of sample and solution reagents. The mixing efficiency is significantly improved compared to passive diffusion, and the reaction is more complete.

[0018] 2. This rapid C-reactive protein (CRP) detection reaction disk utilizes the centrifugal force generated by the disk to achieve a rapid immune response of CRP in biological samples. The operation is simple and convenient, while significantly reducing reaction time. It can be used to improve the efficiency of collecting baseline CRP data from large sample sizes of healthy individuals, enhancing the application value of CRP detection in precision medicine. Attached Figure Description

[0019] Figure 1 This is a schematic diagram of the structure of a reaction disk for rapid detection of C-reactive protein proposed in this utility model;

[0020] Figure 2 This invention provides a reaction disk for rapid detection of C-reactive protein. Figure 1 Enlarged view of section A;

[0021] Figure 3 This is a front view of a reaction disk for rapid detection of C-reactive protein proposed in this utility model;

[0022] Figure 4 This is the standard working curve for C-reactive protein.

[0023] In the diagram: 1. Disc; 2. First liquid storage chamber; 3. Second liquid storage chamber; 4. Third liquid storage chamber; 5. Reaction chamber; 6. Centrifuge chamber; 7. Microfluidic channel; 8. Chamfer; 9. Heating chamber. Detailed Implementation

[0024] The technical solutions of the present utility model will be clearly and completely described below with reference to the accompanying drawings of the embodiments of the present utility model. Obviously, the described embodiments are only some embodiments of the present utility model, and not all embodiments.

[0025] In the description of this utility model, it should be understood that the terms "upper", "lower", "front", "rear", "left", "right", "top", "bottom", "inner", "outer", etc., indicate the orientation or positional relationship based on the orientation or positional relationship shown in the accompanying drawings. They are only for the convenience of describing this utility model and simplifying the description, and do not indicate or imply that the device or element referred to must have a specific orientation, or be constructed and operated in a specific orientation. Therefore, they should not be construed as limitations on this utility model.

[0026] Example:

[0027] like Figures 1-3 A reaction disk for rapid detection of C-reactive protein includes a circular disk 1, which is a carrier of the reaction disk. The center of the bottom of the disk is fixedly connected to the output end of a motor.

[0028] The disk 1 is equipped with a reaction assembly; the reaction assembly includes a first liquid storage chamber 2, a second liquid storage chamber 3, and a reaction chamber 5.

[0029] The first storage chamber 2 has a volume of 135 μL. During the test, a mixed solution of polystyrene microspheres, Tween 20, sodium azide, phosphate buffer, and C-reactive protein polyclonal antibody was added, totaling 125 μL.

[0030] The second storage chamber 3 has a volume of 530 μL. During testing, a mixed solution of phosphate buffer (pH 7.4), Tween 20, polyethylene glycol, sodium chloride, and sodium azide, totaling 500 μL, is added.

[0031] The pH of the phosphate buffer is 7.4.

[0032] The volume of reaction chamber 5 is 640 μL.

[0033] The disk 1 is provided with multiple sets of microfluidic channels 7, the width of which is 0.6-1mm, preferably 0.8mm.

[0034] The first liquid storage tank 2 and the second liquid storage tank 3 are respectively connected to the reaction tank 5 through corresponding microfluidic channels 7; for example Figure 1 and Figure 3 The first liquid storage chamber 2 and the second liquid storage chamber 3 are each connected to the reaction chamber 5 through a microfluidic channel 7.

[0035] like Figure 1 During the test, 2 μL of the sample to be tested is added to the second storage chamber 3. Then, the motor drives the disk 1 to rotate at a centrifugal speed of 200 rpm. After mixing for 15 seconds, the centrifugal speed is increased to 1000 rpm and rotated for 15 seconds. Through centrifugal force, the solutions in the first storage chamber 2 and the second storage chamber 3 are transferred to the reaction chamber 5 through the microfluidic channel 7 for mixing and immune reaction.

[0036] In this immune reaction, highly sensitive and specific C-reactive protein antibodies are directionally coupled to the surface of latex microspheres. After the C-reactive protein in the sample binds to the antibody, an antigen-antibody complex is formed, causing the latex microspheres to aggregate and resulting in a change in actual turbidity. The reaction solution in reaction chamber 5 is taken for spectrophotometric detection, and the absorbance value is recorded. A standard working curve of absorbance versus C-reactive protein concentration is plotted, as shown below. Figure 4 As shown.

[0037] The linear fitting equation for the standard working curve is y = 0.063x + 1.28, R0 2 =0.998.

[0038] The accuracy and repeatability of this testing method were verified using nationally certified reference materials.

[0039] Accuracy: Using the nationally certified standard reference material GBW09228, C-reactive protein solution purity standard, 2 μL of C-reactive protein standard material of known concentration was mixed and reacted through disk 1. After the reaction, the sample was detected by a spectrophotometer. The spectra were observed, the absorbance was recorded, and the measured value of C-reactive protein was calculated using a working curve (rounded to one decimal place). Δc was calculated according to the following formula as the evaluation result of the accuracy of this method.

[0040] Δc=|c 测 -c 理 |

[0041] In the formula:

[0042] c_measured: Measured value, unit mg / L;

[0043] c_theo: theoretical value, unit in mg / L.

[0044] The concentration of C-reactive protein is 54.9 mg / L, with an expanded uncertainty of 2.8 mg / L and k=2. After repeated detection 3 times using the method in the project, the 3 inter-batch measured values obtained via the standard curve are 55.6, 52.7, and 53.1 mg / L respectively, the calculated average value is 53.8 mg / L, and Δc = 1.1 mg / L.

[0045] Repeatability: A standard substance solution of C-reactive protein with a known concentration is added to perform the aforementioned detection and obtain measured values. The relative standard deviation (RSD) is calculated through 6 consecutive tests, which is taken as the result of the repeatability of this method.

[0046]

[0047] In the formula:

[0048] xi: the measured value of the i-th measurement;

[0049] the arithmetic mean of 6 measurements;

[0050] i: measurement sequence number.

[0051] Through 6 tests, the measured values of C-reactive protein are 55.6, 52.7, 53.1, 53.7, 52.1, and 55.9 mg / L respectively, and RSD = 2.86% is calculated by the above formula.

[0052] This method can balance the efficiency, portability and low cost of complex biological sample detection, has important advantages in the fields of point-of-care testing, high-throughput screening and the like, and provides a technical direction for the microfluidic technology to move towards clinical practical application.

[0053] By driving the disk 1 to rotate, there is no need for additional complex mechanical devices such as external pumps and valves. By adjusting the rotation speed of the disk 1, the flow of the solution in the microchannel (such as starting, stopping, and switching paths) can be precisely controlled, so as to promote the uniform mixing of the sample and the solution reagent. The mixing efficiency is significantly improved compared with passive diffusion, and the reaction is sufficient.

[0054] In addition, the centrifugal force can inhibit the residue and adsorption of the solution in the microchannel 7. During the rotation of the disk 1, the centrifugal force is greater than the capillary force and the surface tension of the solution, so that the solution in the microchannel 7 is rapidly transferred, and the reaction efficiency is improved.

[0055] It should be noted that in actual detection, the disk 1 is located in the housing of the detection device, and there is a cover on the top. During detection, that is, when the disk 1 rotates, the cover is closed and fits with the upper surface of the disk 1 to prevent various solutions from splashing out during detection.

[0056] For example Figure 1 and Figure 3 The reaction assembly also includes a third liquid storage chamber 4; the third liquid storage chamber 4 is also connected to the reaction chamber 5 through a corresponding microfluidic channel 7. The third liquid storage chamber 4 is also connected to the reaction chamber 5 through a separate microfluidic channel 7.

[0057] Add 3% (wt / vol%) C18 microsphere methanol solution to the third storage chamber 4.

[0058] Increase the centrifugation speed to 1000 rpm in disc 1 and rotate for 15 seconds. Then, continue to adjust the speed to 1500 rpm and rotate for another 15 seconds. The C18 microspheres in the third storage chamber 4 enter the reaction chamber 5 for mixing and immune reaction to remove proteins and peptides from the sample.

[0059] like Figure 1 and Figure 3 The reaction assembly also includes a centrifuge chamber 6; the centrifuge chamber 6 is also connected to the reaction chamber 5 through a corresponding microfluidic channel 7; the centrifuge chamber 6 is also connected to the reaction chamber 5 through a separate microfluidic channel 7.

[0060] In other words, a reaction assembly has four microfluidic channels 7; the first liquid storage chamber 2, the second liquid storage chamber 3, the third liquid storage chamber 4, and the centrifuge chamber 6 are all connected to the reaction chamber 5 through a microfluidic channel 7.

[0061] like Figure 1 and Figure 3 The first liquid storage chamber 2, the second liquid storage chamber 3, and the third liquid storage chamber 4 are located on the side of the reaction chamber 5 closest to the circular side of the disk 1;

[0062] Centrifuge chamber 6 is located on the side of reaction chamber 5 away from the circular shape of disk 1.

[0063] This ensures that the solution enters the reaction chamber 5 and the centrifuge chamber 6 sequentially through centrifugal force.

[0064] After the reaction, the sample is separated by centrifugation and enters centrifuge chamber 6. It can then be analyzed by large instruments to obtain a cleaner detection background and a better detection signal.

[0065] like Figures 1-3 The connections between the microfluidic channel 7 and the first liquid storage tank 2, the second liquid storage tank 3, and the third liquid storage tank 4 are all designed with chamfers 8.

[0066] The connection between the microfluidic channel 7 and the centrifuge chamber 6 is also designed with a chamfer 8.

[0067] The chamfered 8 design facilitates the flow of solution in and out.

[0068] like Figure 1 and Figure 3The number of reaction components is 2-5, preferably 3.

[0069] Through multiple designs, parallel detection of multiple samples and multiple indicators can be achieved, thereby improving detection efficiency.

[0070] like Figures 1-3 The disc 1 is also equipped with a heating chamber 9; the second liquid storage chamber 3 is located inside the heating chamber 9, and the heating chamber 9 and the second liquid storage chamber 3 are separated by a partition. The partition is thermally conductive and heats the solution in the second liquid storage chamber 3.

[0071] A heating component, such as a heating wire, is installed inside the heating chamber 9 to heat the solution in the second storage chamber 3 to 37°C, thereby accelerating the subsequent reaction.

[0072] The rapid C-reactive protein (CRP) detection reaction disk utilizes the centrifugal force generated by disk 1 to achieve a rapid immune response of CRP in biological samples. The operation is simple and convenient, while significantly reducing reaction time. It can be used to improve the efficiency of collecting baseline CRP data from large sample sizes of healthy individuals, enhancing the application value of CRP detection in precision medicine.

[0073] The above description is only a preferred embodiment of the present utility model, but the protection scope of the present utility model is not limited thereto. Any equivalent substitutions or changes made by those skilled in the art within the technical scope disclosed in the present utility model, based on the technical solution and the inventive concept of the present utility model, should be included within the protection scope of the present utility model.

Claims

1. A reaction disk for rapid detection of C-reactive protein, comprising a disk (1), characterized in that, The disk (1) is equipped with a reaction assembly; The reaction assembly includes a first liquid storage tank (2), a second liquid storage tank (3), and a reaction tank (5); The disk (1) is provided with multiple sets of microfluidic channels (7); The first liquid storage chamber (2) and the second liquid storage chamber (3) are respectively connected to the reaction chamber (5) through corresponding microfluidic channels (7).

2. The reaction disk for rapid detection of C-reactive protein according to claim 1, characterized in that, The reaction assembly also includes a third liquid storage tank (4); The third liquid storage chamber (4) is also connected to the reaction chamber (5) through the corresponding microfluidic channel (7).

3. The reaction disk for rapid detection of C-reactive protein according to claim 2, characterized in that, The reaction assembly also includes a centrifuge chamber (6); The centrifuge chamber (6) is also connected to the reaction chamber (5) through a corresponding microfluidic channel (7).

4. The reaction disk for rapid detection of C-reactive protein according to claim 1, characterized in that, The width of the microfluidic channel (7) is 0.6-1 mm.

5. The reaction disk for rapid detection of C-reactive protein according to claim 2, characterized in that, The connection points between the microfluidic channel (7) and the first liquid storage tank (2), the second liquid storage tank (3), and the third liquid storage tank (4) are all designed with chamfers (8).

6. A reaction disk for rapid detection of C-reactive protein according to any one of claims 1-5, characterized in that, The number of reaction components is 2-5.

7. A reaction disk for rapid detection of C-reactive protein according to claim 6, characterized in that, The number of reaction components is 3.

8. The reaction disk for rapid detection of C-reactive protein according to claim 1, characterized in that, The disc (1) is also provided with a heating chamber (9); The second liquid storage chamber (3) is located inside the heating chamber (9).

Images