Tumor necrosis factor alpha detection kit using chemiluminescence immunoassay technology

Abstract

The invention discloses a tumor necrosis factor alpha detection kit using a chemiluminescence immunoassay technology, and relates to the technical field of detection kits. An anti-human tumor necrosis factor alpha antibody is used for coating a microporous ELISA plate, after a standard substance is added, the tumor necrosis factor alpha can be tightly combined with the coating antibody, then a detection antibody is added to form a stable antibody-antigen-enzyme-labeled antibody compound, free components are thoroughly removed through plate washing operation, then a chromogenic substrate TMB is added, and the detection sensitivity is high. The TMB is blue under the catalysis of a detection antibody, finally, the TMB is converted into yellow after a stop buffer is added, the depth of the color is positively correlated with the concentration of the tumor necrosis factor alpha in a sample, the absorbance is measured through an enzyme-linked immunosorbent assay instrument, the concentration of the sample can be accurately calculated, and the fixed seat can be prevented from shifting under the cooperation of a connecting shaft and a roller; and by arranging the bearing mechanism, the purposes of stretching out and storing the fixing mechanism can be achieved, and taking, placing and protection are facilitated.

Description

Technical Field

[0001] The present invention relates to the technical field of detection kits, and particularly relates to a tumor necrosis factor α detection kit using chemiluminescent immunoassay technology. Background Art

[0002] Tumor necrosis factor is the earliest discovered cytokine that can cause hemorrhagic necrosis of tumor tissue cells. It is mainly secreted by macrophages and adipocytes in adipose tissue and is named because it can directly kill tumor cells in vivo and in vitro. According to its source and structure, it is mainly divided into TNF-α and TNF-β. TNF-α is mainly produced by activated monocytes-macrophages and many other cells. High-dose TNF-α can cause cachexia, and patients show progressive emaciation. Therefore, TNF-α is also called cachectin.

[0003] The methods for measuring tumor necrosis factor α are mainly divided into three categories: bioactivity detection method, immunological detection method, and molecular biology detection method. Bioactivity detection method: Currently, the MTT colorimetric method in the cell proliferation method is more commonly used. The MTT method has poor specificity and the test samples are easily interfered by other active substances. For example, the activities of TNF and IL-1 are cross-reactive, resulting in deviation of the results. In the immunological detection method, the commonly used enzyme-linked immunosorbent assay technology is currently the main detection means for tumor necrosis factor α. However, the enzyme-labeled substances in the ELISA method are unstable and are easily affected by the environment such as temperature, pH value, etc. The method has poor repeatability and sensitivity. The component detected by the molecular biology detection method is the mRNA of cytokines, which cannot objectively reflect the protein expression level and causes differences in the results. Therefore, we propose a tumor necrosis factor α detection kit using chemiluminescent immunoassay technology. Summary of the Invention

[0004] The purpose of the present invention is to provide a tumor necrosis factor α detection kit using chemiluminescent immunoassay technology to solve the problems mentioned in the above background art.

[0005] The present invention specifically adopts the following technical solutions to achieve the above purpose:

[0006] A tumor necrosis factor α detection kit using chemiluminescent immunoassay technology, comprising:

[0007] A box body, which is used to protect the internal components;

[0008] Fixing mechanism, the fixing mechanism is arranged inside the box body, the fixing mechanism includes five fixing seats uniformly arranged longitudinally along the box body, and inside each fixing seat, standard products, sample diluents, detection antibodies, washing buffers, termination liquids, sealing plates and microplate enzyme-linked immunosorbent assay (ELISA) plates are uniformly arranged horizontally. Fixing frames are detachably installed on both sides of the fixing seat, and three connecting shafts are uniformly arranged longitudinally on the outer side of each fixing frame, and rollers are sleeved on the circumferential surface of each connecting shaft;

[0009] Supporting mechanism, the supporting mechanism is arranged below the fixing mechanism, the supporting mechanism includes a supporting plate, and two symmetrically arranged electric push rods are installed at the lower end of the supporting plate;

[0010] Sealing plate, the sealing plate is detachably installed above the box body.

[0011] Further, the concentrations of the standard products from front to back are successively 40, 20, 10, 5, 2.5 pg / ml from high to low.

[0012] Further, through holes are opened inside the fixing seat, the standard products, the sample diluents, the detection antibodies, the washing buffers, the termination liquids and the sealing plates are installed in the through holes, and a through groove is opened inside the right side of the fixing seat, and the microplate ELISA plates are installed in the through groove.

[0013] Further, installation grooves are opened at the inner ends of the fixing frames, and both sides of the fixing seat are installed in the installation grooves.

[0014] Further, positioning seats are installed at the outer ends of the fixing frames, and empty grooves are opened at the outer sides of the positioning seats. The rollers are installed in the empty grooves, and limiting grooves are opened on the inner side walls of the box body. The rollers are movably connected to the box body through the limiting grooves.

[0015] Further, three rows of top seats are uniformly arranged longitudinally at the lower end of the supporting plate. Below each top seat, a supporting plate is movably connected through a mounting shaft, the supporting plate is movably connected to a moving seat through the mounting shaft, and the moving seats on the same side are connected through a limiting shaft. Snap rings are installed on the circumferential surfaces at both ends of each limiting shaft.

[0016] Further, empty grooves are opened at both ends of the supporting plate, and the top seats and the moving seats are installed in the empty grooves.

[0017] Further, moving grooves are opened on the inner bottom wall of the box body. The lower ends of the moving seats are movably connected to the box body through the moving grooves, and positioning grooves with arc-shaped structures at both ends are opened on the front and back of the moving grooves. The limiting shafts are movably connected to the box body through the positioning grooves.

[0018] Furthermore, connection blocks are installed at the upper parts on both sides of the box body, connection grooves are formed at the lower end of the sealing plate, and the connection blocks are movably connected with the connection grooves.

[0019] Furthermore, a mounting seat is installed at the upper end of the sealing plate, rotating shafts are detachably installed on both sides of the mounting seat, and an anti-slip cylinder is sleeved on the circumferential surface of the rotating shafts.

[0020] The beneficial effects of the present invention are as follows:

[0021] 1. The present invention coats a microplate with anti-human tumor necrosis factor α antibody. When the standard product is added, the tumor necrosis factor α therein will tightly bind to the coated antibody. Subsequently, the detection antibody is added to form a stable antibody-antigen-enzyme-labeled antibody complex. After the plate washing operation, the free components are completely removed. Then, the chromogenic substrate TMB is added. Under the catalysis of the detection antibody, TMB presents blue. Finally, after adding the termination solution, it turns yellow. The depth of the color is positively correlated with the concentration of tumor necrosis factor α in the sample. By measuring the absorbance with an enzyme-labeled instrument at a wavelength of 450 nm, the sample concentration can be accurately calculated. When the fixed seat moves vertically inside the box body, under the action of the connecting shaft, the rollers will move inside the box body, that is, it can avoid the deviation of the standard product, sample diluent, detection antibody, washing buffer, termination solution, sealing film and microplate, so as to ensure the actual detection effect.

[0022] 2. The present invention can achieve the purpose of supporting and height adjustment of the supporting plate under the action of the electric push rod. When in use, the sealing plate is removed, and the electric push rod is started, so that the fixing mechanism can extend out of the box body for easy use. After use, the electric push rod contracts, so that the fixing mechanism is received inside the box body, and then the sealing plate is installed to protect the internal components. When the distance between the supporting plate and the inner bottom wall of the box body changes, the top seat will move vertically. Under the action of the mounting shaft, the included angle between the supporting plate and the top seat and the moving seat will change. At this time, under the action of the limiting shaft, the moving seat will move horizontally along the inner bottom wall of the box body, so as to further improve the use stability of the supporting plate. Brief Description of the Drawings

[0023] Figure 1 is a three-dimensional schematic diagram of the present invention;

[0024] Figure 2 is a front sectional view of the present invention;

[0025] Figure 3 is a three-dimensional schematic diagram of the fixing mechanism in the present invention;

[0026] Figure 4 is a three-dimensional schematic diagram of the supporting mechanism in the present invention;

[0027] Figure 5 It is an exploded perspective view of the box body and the sealing plate in the present invention.

[0028] Reference numerals: 10, box body; 20, fixing mechanism; 21, fixing seat; 22, standard product; 23, sample diluent; 24, detection antibody; 25, washing buffer; 26, termination solution; 27, sealing film; 28, microplate enzyme immunoassay plate; 29, fixing frame; 210, connecting shaft; 211, roller; 30, supporting mechanism; 31, supporting plate; 32, electric push rod; 33, top seat; 34, mounting shaft; 35, supporting plate; 36, moving seat; 37, limiting shaft; 38, snap ring; 40, sealing plate; 50, mounting seat; 60, rotating shaft. Detailed implementation manners

[0029] To make the objectives, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings in the embodiments of the present invention.

[0030] Please refer to Figure 1 - Figure 5 , the present invention provides a tumor necrosis factor α detection kit using chemiluminescence immunoassay technology, including:

[0031] A box body 10 for protecting the internal components.

[0032] A fixing mechanism 20 is arranged inside the box body 10. The fixing mechanism 20 includes five fixing seats 21 uniformly arranged longitudinally along the box body 10. Inside each fixing seat 21, a standard product 22, a sample diluent 23, a detection antibody 24, a washing buffer 25, a termination solution 26, a sealing film 27 and a microplate enzyme immunoassay plate 28 are uniformly arranged horizontally. Fixing frames 29 are detachably installed on both sides of the fixing seat 21. Three connecting shafts 210 are uniformly arranged longitudinally on the outer side of each fixing frame 29, and a roller 211 is sleeved on the circumferential surface of each connecting shaft 210.

[0033] Under the action of the fixing seat 21, the installation stability of the standard product 22, the sample diluent 23, the detection antibody 24, the washing buffer 25, the termination solution 26, the sealing film 27 and the microplate enzyme immunoassay plate 28 can be realized. Under the action of the fixing frame 29, the installation stability of the fixing seat 21 can be realized, and the installation stability of the connecting shaft 210 and the roller 211 on its circumferential surface can also be ensured.

[0034] Coat a microplate enzyme immunoassay plate 28 with an anti-human tumor necrosis factor α antibody. When the standard 22 is added, the tumor necrosis factor α therein will bind tightly to the coated antibody. Subsequently, add the detection antibody 24 to form a stable antibody-antigen-enzyme-labeled antibody complex. After the plate washing operation, the free components are completely removed. Then add the chromogenic substrate TMB. Under the catalysis of the detection antibody 24, TMB shows blue. Finally, after adding the stop solution 26, it turns yellow. The depth of the color is positively correlated with the concentration of tumor necrosis factor α in the sample. By measuring the absorbance with an enzyme immunoassay reader at a wavelength of 450 nm, the sample concentration can be accurately calculated. When the fixing base 21 moves vertically inside the box body 10, under the action of the connecting shaft 210, the roller 211 will move inside the box body 10, which can avoid the deviation of the standard 22, sample diluent 23, detection antibody 24, washing buffer 25, stop solution 26, sealing film 27 and microplate enzyme immunoassay plate 28, thus ensuring the actual detection effect.

[0035] A supporting mechanism 30 is provided below the fixing mechanism 20. The supporting mechanism 30 includes a supporting plate 31, and two symmetrically arranged electric push rods 32 are installed at the lower end of the supporting plate 31.

[0036] Under the action of the supporting plate 31, the purpose of supporting the fixing mechanism 20 can be achieved. Under the action of the electric push rod 32, the purpose of supporting and height adjustment of the supporting plate 31 can be achieved. When in use, remove the sealing plate 40 and start the electric push rod 32, which can make the fixing mechanism 20 extend out of the box body 10 for easy use. After use, the electric push rod 32 contracts, making the fixing mechanism 20 retract into the box body 10, and then install the sealing plate 40 to protect the internal components.

[0037] A sealing plate 40 is detachably installed above the box body 10.

[0038] Under the action of the sealing plate 40, the sealing performance of the box body 10 can be ensured, and the internal components can be prevented from falling off.

[0039] In this embodiment, preferably, the concentrations of the standard 22 from front to back are 40, 20, 10, 5, 2.5 pg / ml from high to low; it can meet the usage requirements under different conditions and improve the applicability.

[0040] In this embodiment, preferably, a through hole is provided inside the fixing base 21, and the standard product 22, sample diluent 23, detection antibody 24, washing buffer 25, termination solution 26 and sealing plate film 27 are installed in the through hole. Moreover, a through groove is provided inside the right side of the fixing base 21, and the microplate 28 is installed in the through groove. By providing the through hole, the installation purpose of the standard product 22, sample diluent 23, detection antibody 24, washing buffer 25, termination solution 26 and sealing plate film 27 can be ensured. By providing the through groove, the installation purpose of the microplate 28 can be ensured.

[0041] In this embodiment, preferably, an installation groove is provided at the inner end of the fixing frame 29, and both sides of the fixing base 21 are installed in the installation groove. Under the action of the installation groove, the connection stability between the fixing base 21 and the fixing frame 29 can be ensured.

[0042] In this embodiment, preferably, a positioning seat is installed at the outer end of the fixing frame 29, and an empty groove is provided on the outer side of the positioning seat. The roller 211 is installed in the empty groove, and a limiting groove is provided on the inner side wall of the box body 10. The roller 211 is movably connected to the box body 10 through the limiting groove. Under the action of the empty groove, space can be provided for the rotation of the roller 211. Under the action of the limiting groove, the moving path of the roller 211 can be limited.

[0043] In this embodiment, preferably, three rows of top seats 33 are uniformly arranged along the longitudinal direction at the lower end of the supporting plate 31. A supporting plate 35 is movably connected to the lower side of each top seat 33 through a mounting shaft 34. The supporting plate 35 is movably connected to a moving seat 36 through the mounting shaft 34. The moving seats 36 on the same side are connected through a limiting shaft 37. Snap rings 38 are installed on the circumferential surfaces at both ends of each limiting shaft 37. Under the action of the supporting plate 31, the installation stability of the top seats 33 can be ensured. Under the action of the mounting shaft 34, the connection stability between the supporting plate 35 and the top seats 33 and the moving seats 36 can be ensured. Under the action of the limiting shaft 37, the connection stability of the moving seats 36 on the same side can be ensured. Under the action of the snap rings 38, the limiting shaft 37 can be prevented from falling off. When the distance between the supporting plate 31 and the inner bottom wall of the box body 10 changes, the top seats 33 will move in the vertical direction. Under the action of the mounting shaft 34, the included angles between the supporting plate 35 and the top seats 33 and the moving seats 36 will change. At this time, under the action of the limiting shaft 37, the moving seats 36 will move horizontally along the inner bottom wall of the box body 10, thereby further improving the use stability of the supporting plate 31.

[0044] In this embodiment, preferably, empty grooves are provided at both ends of the supporting plate 35, and the top seats 33 and the moving seats 36 are installed in the empty grooves. Under the cooperation of the empty grooves and the mounting shaft 34, space can be provided for the angle adjustment at both ends of the supporting plate 35.

[0045] In this embodiment, preferably, a moving groove is formed in the inner bottom wall of the box body 10. The lower end of the moving seat 36 is movably connected to the box body 10 through the moving groove. The front and back surfaces of the moving groove are provided with positioning grooves with arc-shaped structures at both ends. The limiting shaft 37 is movably connected to the box body 10 through the positioning grooves. By providing the moving groove, space can be provided for the movement of the lower end of the moving seat 36. By providing the positioning grooves, space can be provided for the movement of the limiting shaft 37, and a limiting effect is also achieved.

[0046] In this embodiment, preferably, connection blocks are installed on the upper parts of both sides of the box body 10. Connection grooves are formed in the lower end of the sealing plate 40. The connection blocks are movably connected to the connection grooves. The connection grooves can ensure the installation stability of the connection blocks, and the cooperation between the connection blocks and the connection grooves can achieve the disassembly of the sealing plate 40 and the box body 10.

[0047] In this embodiment, preferably, a mounting seat 50 is installed on the upper end of the sealing plate 40. The two sides of the mounting seat 50 are detachably installed with rotating shafts 60. The circumferential surface of the rotating shaft 60 is sleeved with an anti-slip cylinder. The sealing plate 40 can support the mounting seat 50. The mounting seat 50 can ensure the installation stability of the rotating shaft 60. The rotating shaft 60 is movably connected to the mounting seat 50, that is, the parallel and perpendicular states of the rotating shaft 60 and the mounting seat 50 can be achieved. When in the vertical state, the overall lifting purpose can be achieved. The anti-slip cylinder can increase the friction force and ensure the use effect.

[0048] Working principle and usage process of the present invention: The box body 10 is used to protect the internal components; the microplate enzyme immunoassay plate 28 is coated with anti-human tumor necrosis factor α antibody. When the standard product 22 is added, the tumor necrosis factor α therein will tightly bind to the coated antibody. Subsequently, the detection antibody 24 is added to form a stable antibody-antigen-enzyme-labeled antibody complex. After the plate washing operation, the free components are completely removed. Then, the chromogenic substrate TMB is added. Under the catalysis of the detection antibody 24, TMB presents blue. Finally, after adding the stop solution 26, it turns yellow. The depth of the color is positively correlated with the concentration of tumor necrosis factor α in the sample. By measuring the absorbance with an enzyme-labeled instrument at a wavelength of 450 nm, the sample concentration can be accurately calculated. When the fixed seat 21 moves vertically inside the box body 10, under the action of the connecting shaft 210, the roller 211 will move inside the box body 10, that is, it can prevent the standard product 22, sample diluent 23, detection antibody 24, washing buffer 25, stop solution 26, sealing film 27, and microplate enzyme immunoassay plate 28 from shifting, thereby ensuring the actual detection effect; under the action of the supporting plate 31, the supporting purpose of the fixing mechanism 20 can be achieved. Under the action of the electric push rod 32, the supporting and height adjustment purposes of the supporting plate 31 can be achieved. When in use, the sealing plate 40 is removed, and the electric push rod 32 is started, so that the fixing mechanism 20 can extend out of the box body 10 for easy use. After use, the electric push rod 32 contracts, so that the fixing mechanism 20 is received inside the box body 10, and then the sealing plate 40 is installed to protect the internal components. When the distance between the supporting plate 31 and the inner bottom wall of the box body 10 changes, the top seat 33 will move vertically. Under the action of the mounting shaft 34, the included angle between the supporting plate 35 and the top seat 33 and the moving seat 36 will change. At this time, under the action of the limiting shaft 37, the moving seat 36 will move horizontally along the inner bottom wall of the box body 10, thereby further improving the use stability of the supporting plate 31; under the action of the sealing plate 40, the sealing performance of the box body 10 can be ensured, preventing the internal components from falling off. Under the action of the sealing plate 40, the supporting purpose of the mounting seat 50 can be achieved. Under the action of the mounting seat 50, the installation stability of the rotating shaft 60 can be ensured, and the rotating shaft 60 is movably connected to the mounting seat 50, that is, the parallelism and perpendicularity of the rotating shaft 60 and the mounting seat 50 can be achieved. When in the vertical state, the overall lifting purpose can be achieved. Under the action of the anti-slip cylinder, the friction force can be increased to ensure the use effect.

[0049] The foregoing description of the disclosed embodiments enables those skilled in the art to practice or use the present invention. Various modifications to these embodiments will be readily apparent to those skilled in the art, and the general principles defined herein may be implemented in other embodiments without departing from the spirit or scope of the present invention. Thus, the present invention is not intended to be limited to the embodiments shown herein but is to be accorded the widest scope consistent with the principles and novel features disclosed herein.

Claims

1. A tumor necrosis factor α detection kit using chemiluminescence immunoassay technology, characterized in that, Comprising: A box body (10) for protecting internal components. A fixing mechanism (20) disposed inside the box body (10). The fixing mechanism (20) includes five fixing seats (21) uniformly arranged longitudinally along the box body (10). Inside each fixing seat (21), standards (22), sample diluent (23), detection antibody (24), washing buffer (25), termination solution (26), sealing film (27), and microplate enzyme immunoassay plate (28) are uniformly arranged horizontally. Fixing frames (29) are detachably installed on both sides of the fixing seat (21). On the outer side of each fixing frame (29), three connecting shafts (210) are uniformly arranged longitudinally. A roller (211) is sleeved on the circumferential surface of each connecting shaft (210). A supporting mechanism (30) disposed below the fixing mechanism (20). The supporting mechanism (30) includes a supporting plate (31), and two symmetrically arranged electric push rods (32) are installed at the lower end of the supporting plate (31). A sealing plate (40) detachably installed above the box body (10).

2. The tumor necrosis factor α detection kit using chemiluminescence immunoassay technology according to claim 1, wherein: The concentrations of the standards (22) from front to back are successively 40, 20, 10, 5, 2.5 pg / ml.

3. A tumor necrosis factor α detection kit using chemiluminescent immunoassay technology according to claim 1, characterized in that: Through holes are formed inside the fixing seat (21). The standards (22), the sample diluent (23), the detection antibody (24), the washing buffer (25), the termination solution (26), and the sealing film (27) are installed in the through holes. A through groove is formed inside the right side of the fixing seat (21), and the microplate enzyme immunoassay plate (28) is installed in the through groove.

4. A tumor necrosis factor α detection kit using chemiluminescence immunoassay technology according to claim 1, characterized in that: Installation grooves are formed at the inner ends of the fixing frames (29), and both sides of the fixing seat (21) are installed in the installation grooves.

5. A tumor necrosis factor α detection kit using chemiluminescence immunoassay technology according to claim 1, characterized in that: Positioning seats are installed at the outer ends of the fixing frames (29), and empty grooves are formed on the outer sides of the positioning seats. The rollers (211) are installed in the empty grooves. Limiting grooves are formed on the inner side walls of the box body (10), and the rollers (211) are movably connected to the box body (10) through the limiting grooves.

6. A tumor necrosis factor α detection kit using chemiluminescent immunoassay technology according to claim 1, characterized in that: Three rows of top seats (33) are uniformly arranged longitudinally at the lower end of the supporting plate (31). A supporting plate (35) is movably connected to the lower side of each top seat (33) through a mounting shaft (34). The supporting plate (35) is movably connected to a moving seat (36) through the mounting shaft (34). The moving seats (36) on the same side are connected by a limiting shaft (37). Snap rings (38) are installed on the circumferential surfaces at both ends of each limiting shaft (37).

7. A kit for detecting tumor necrosis factor α using chemiluminescent immunoassay technology according to claim 6, characterized in that: Empty grooves are formed at both ends of the supporting plate (35), and the top seats (33) and the moving seats (36) are installed in the empty grooves.

8. A kit for detecting tumor necrosis factor-α using chemiluminescence immunoassay technology according to claim 6, characterized in that: A moving groove is formed in the inner bottom wall of the box body (10). The lower end of the moving seat (36) is movably connected to the box body (10) through the moving groove. Positioning grooves with arc-shaped structures at both ends are formed in the front and back of the moving groove. The limiting shaft (37) is movably connected to the box body (10) through the positioning grooves.

9. A tumor necrosis factor α detection kit using chemiluminescence immunoassay technology according to claim 1, characterized in that: Connecting blocks are installed on the upper parts of both sides of the box body (10). Connecting grooves are formed at the lower ends of the sealing plates (40). The connecting blocks are movably connected to the connecting grooves.

10. A tumor necrosis factor α detection kit using chemiluminescent immunoassay technology according to claim 1, characterized in that: An installation seat (50) is installed at the upper end of the sealing plate (40). Rotating shafts (60) are detachably installed on both sides of the installation seat (50). Anti-slip cylinders are sleeved on the circumferential surfaces of the rotating shafts (60).

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