Three-chromatography-channel cancer early screening test strip based on exosome protein

Abstract

The utility model discloses a three-chromatography-channel cancer early screening test strip based on exosome protein. The three-chromatography-channel cancer early screening test strip comprises a drainage sheet, the first chromatography channel, the second chromatography channel and the third chromatography channel are arranged around the drainage piece; the first chromatography channel, the second chromatography channel and the third chromatography channel respectively comprise a sample pad, a chromatography film, a water absorption pad and a bottom plate; the chromatographic membrane is coated with the colloidal gold pad, the detection line and the quality control line; the colloidal gold pads of the three chromatography channels are coated with three kinds of gold-labeled antibodies containing different exosome proteins; the detection line is coated with an antibody which is specifically combined with a substance to be detected; and the quality control lines are coated with antibodies which are specifically combined with the corresponding colloidal gold particles. The test strip can be used for detecting three special transmembrane proteins CD63, CD81 and CD9 in the exosome, and further can be used for identifying circulating tumor cells such as gastric cancer, lung cancer, lymph cancer and colorectal cancer. Early screening and early discovery of specific cancer cells can be realized, postoperative rehabilitation monitoring is also facilitated, and the method has a great clinical application space.

Description

Technical Field

[0001] The utility model belongs to the technical field of medical devices, and particularly relates to a three-chromatographic channel composite test strip for early cancer screening based on exosome proteins and a preparation method thereof. Background Art

[0002] There are many types of traditional test strips that can be sorted based on size, complexity, and specific binding of antigens and antibodies. However, these traditional test strips have the following problems in clinical applications:

[0003] (1) Limited test items: Traditional test strips can usually only detect a single or a few indicators, and the test items are relatively simple;

[0004] (2) The disadvantage of single-channel paper chromatography test strips is that the detection efficiency is low and it cannot meet the requirements of multi-item testing of large quantities of samples.

[0005] In addition to the above problems, the current multi-channel test strips can only realize multiple tests on different samples, resulting in multiple sampling required when different types of tests are needed. It is impossible to perform multiple tests on the same sample at the same time, and it is even impossible to accurately compare multiple tests.

[0006] Therefore, there is an urgent need for an efficient test paper that can simultaneously perform multiple types of tests on the same sample and perform accurate comparisons. Utility Model Content

[0007] In response to the above technical problems, the present invention aims to provide a low-cost, simple and efficient three-layer chromatography channel composite test strip for early cancer screening based on exosome proteins.

[0008] The technical solutions provided by this utility model are as follows:

[0009] The three-layered cancer early screening test strip based on exosome proteins includes:

[0010] drainage flap; and,

[0011] a first, a second, and a third chromatography channel disposed around the drainage plate;

[0012] The first, second and third chromatography channels each include a sample pad, a chromatography membrane, an absorbent pad and a bottom plate;

[0013] The chromatography membrane is coated with a colloidal gold pad, a detection line and a quality control line;

[0014] The colloidal gold pads of the three chromatography channels are coated with three gold-labeled antibodies containing different exosomes, one of which is coated on each chromatography channel;

[0015] The test line is coated with an antibody that specifically binds to the substance to be tested;

[0016] The quality control lines are coated with antibodies that specifically bind to the corresponding colloidal gold particles.

[0017] In a possible implementation, the drainage sheet is sequentially connected to a sample pad, a chromatography membrane, and a water-absorbing pad supported by a bottom plate.

[0018] In a possible implementation, the colloidal gold pad, the detection line, and the quality control line are separated by a chromatography membrane.

[0019] Furthermore, the distances between the colloidal gold pad, the detection line and the quality control line, as well as their setting positions, remain consistent in the three channels.

[0020] Furthermore, the size, thickness and water absorption properties of the water-absorbing material are consistent in the three chromatography channels.

[0021] In a possible implementation, the gold-labeled antibody is formed by combining gold particles with an exoprotein antibody.

[0022] Furthermore, the gold-labeled antibody for the exoprotein is selected from one of gold-labeled Anti-CD63, gold-labeled Anti-CD81 and gold-labeled Anti-CD9.

[0023] In a possible implementation, the chromatography membrane is a nitrocellulose membrane; and the bottom plate is a PVC plate.

[0024] In a possible implementation, the first, second, and third chromatography channels are uniformly connected to the drainage plate.

[0025] In a possible implementation, the drainage plate is provided with a rear injection port.

[0026] The beneficial effects of the utility model are as follows:

[0027] (1) One sampling can be used to perform three tests, greatly reducing the demand for samples;

[0028] (2) Simplified sample pretreatment: The optimized test strips only require simple pretreatment before testing, which simplifies the testing process and makes the test more convenient and quick;

[0029] (3) The multi-channel test strip is used to detect three exosome outer membrane proteins, CD63, CD81, and CD9, requiring less sample, greatly improving the accuracy of early screening and also facilitating rehabilitation testing for patients;

[0030] (4) The multi-channel test strip can achieve more accurate comparison of different types of test results through structural design, providing more research and analysis dimensions;

[0031] (5) The present invention can realize the screening of different cancers, such as lymphoma, lung cancer and colorectal cancer, and has important significance in the field of life sciences. BRIEF DESCRIPTION OF THE DRAWINGS

[0032] Figure 1 It is a schematic diagram of the three-dimensional structure of the present invention;

[0033] Figure 2 It is a schematic diagram of a bird's-eye view of the present invention;

[0034] Figure 3 This is a schematic diagram of the relative positions of individual test strips of the three-layer chromatography channel test strip of the present invention;

[0035] FIG4 is a diagram showing the simulated effect of sample detection under the action of chromatographic force according to the present invention; (a) sample addition diagram; (b) antigen-gold labeled antibody binding diagram; (c) detection line effect diagram; (d) quality control line effect diagram;

[0036] In the figure: 1-inlet, 2-bottom plate, 3-drainage sheet, 4-sample pad, 5-colloidal gold pad, 6-chromatographic membrane, 7-test line, 8-quality control line, 9-absorbent pad. DETAILED DESCRIPTION

[0037] The present invention will be further described below with reference to the accompanying drawings, but the present invention is not limited thereto.

[0038] Example

[0039] like Figures 1 to 3 As shown, the three-layer chromatography channel cancer early screening test strip based on exosome proteins includes:

[0040] Drainage sheet 3; and

[0041] First, second and third chromatography channels arranged around the drainage plate 3;

[0042] The first, second and third chromatography channels each include a sample pad 4, a chromatography membrane 6, a water absorbent pad 9 and a bottom plate 2;

[0043] The chromatography membrane 6 is coated with a colloidal gold pad 5, a detection line 7 and a quality control line 8;

[0044] The colloidal gold pads 5 of the three chromatography channels are coated with three gold-labeled antibodies containing different exosome proteins, one of which is coated on each chromatography channel;

[0045] The detection line 7 is coated with an antibody that specifically binds to the substance to be detected;

[0046] The quality control lines 8 are coated with antibodies that specifically bind to the corresponding colloidal gold particles.

[0047] In a possible implementation, the drainage sheet 3 is sequentially connected to the sample pad 4 , the chromatography membrane 6 , and the absorbent pad 9 supported by the bottom plate 2 .

[0048] In a possible implementation, the colloidal gold pad 5 , the detection line 7 and the quality control line 8 are separated by a chromatography membrane.

[0049] As a specific embodiment, the distances and the setting positions among the colloidal gold pad 5 , the detection line 7 and the quality control line 8 are kept consistent in the three channels.

[0050] It can be understood that the distances and settings between the colloidal gold pad 5, the detection line 7 and the quality control line 8 are kept consistent in the three channels in order to keep the chromatographic distances consistent during the detection process and to facilitate intuitive comparison of the control variables.

[0051] As a specific embodiment, the size, thickness and water absorption performance of the absorbent pad 9 are consistent in the three chromatography channels. Similarly, this is also for the purpose of controlling variables.

[0052] In a possible implementation, the gold-labeled antibody is formed by combining gold particles with an exoprotein antibody.

[0053] As a specific embodiment, the gold-labeled antibody for the exocytic protein is selected from one of gold-labeled Anti-CD63, gold-labeled Anti-CD81 and gold-labeled Anti-CD9.

[0054] In a possible implementation, the chromatography membrane 6 is a nitrocellulose membrane; and the bottom plate 2 is a PVC plate.

[0055] It should be noted that the test strip can be a drainage piece and three channels placed on a flat plate, or the bottom plate 2 can be expanded into a disc to form an integral structure.

[0056] In a possible implementation, the first, second, and third chromatography channels are uniformly connected to the drainage plate 3 .

[0057] In a possible implementation, the drainage plate 3 is provided with a rear injection port 1 .

[0058] As a specific embodiment, the base plate 2 of the entire test strip is a 75mm diameter disc, and the drain plate has a diameter of 25mm. The width of each of the three chromatographic channels is 5mm, and the angle between the three chromatographic channels is 120°. The injection hole has a diameter of 5mm x 5mm.

[0059] As a specific embodiment, the detection line 7 is coated with antibodies Anti-CD63, Anti-CD81 and Anti-CD9 that specifically bind to the substance to be detected.

[0060] As a specific embodiment, the quality control line 8 is coated with antibodies Anti-CD63, Anti-CD81 and Anti-CD9 that specifically bind to their respective corresponding colloidal gold particles.

[0061] How to use the test strips:

[0062] Within 1 hour of use, remove the three-layer chromatography channel test strip from the low-temperature environment, add an appropriate amount of sample, and place the disc flat to ensure that the drainage piece is evenly distributed. After a period of time, observe the changes in the detection lines and quality control lines of the three test strips.

[0063] As shown in Figure 4, the gold-labeled antibody in the colloidal gold pad of the present invention is obtained by combining gold particles with antigen-labeled antibodies through Au-S bonds. It is captured by non-labeled antibodies at the test line (T line) to form a "colloidal gold antibody-viral antigen-T line antibody" complex, and the excess gold-labeled antibody will flow to the absorbent pad. When it flows to the quality control line (C line), it will be captured by the antibodies at the quality control line to form a "gold-labeled antibody-C line antibody" complex. At the same time, colloidal gold accumulates in large quantities, and both the test line and the quality control line appear red, thereby achieving the purpose of detection. The single-hole three-channel configuration is conducive to the identification of circulating tumor cells such as gastric cancer, lung cancer, lymphoma and stone cancer. The circular drainage plate ensures that when the sample is added uniformly, it can ensure that the sample flows evenly into each test paper slot, so that it can be evenly chromatographed at the same time. The single sample addition hole reduces the number of samples required for detection, reduces the number of sampling times, and is beneficial to the patient's physical recovery. The application of three chromatography channels improves efficiency while greatly reducing equipment costs. Different detection lines correspond to different circulating tumor cells and also to different lesions. The combination of test strips can be adjusted quickly and conveniently according to clinical needs. The three-layer chromatography channel composite test strips for early screening of specific circulating tumor cells (CTCs) using exosome proteins are highly efficient, highly sensitive, and highly feedback-free. They do not harm the health of patients and are beneficial for early screening and rehabilitation testing of patients. Detection of three special transmembrane proteins, CD63, CD81, and CD9, in exosomes is beneficial for the identification of circulating tumor cells such as gastric cancer, lung cancer, lymphoma, and stone cancer. The utility model can not only realize the early screening and early detection of specific cancers, but also facilitate postoperative rehabilitation monitoring, and has great room for development in future clinical applications.

[0064] The three-channel chromatographic test strips described in this utility model significantly improve the detection rate of target antigens, utilizing multiple sets of antigen-antibody specific binding to capture antigens. The gold-labeled antibody in the colloidal gold pad is prepared through the synthesis of "Au-S" bonds. The antigen and gold-labeled antibody bind to the test line through chromatography, completing the secondary capture of the antibody and antigen at the immunological level and converting them into a visible color medium. Through the triple detection of Anti-CD63, Anti-CD81, and Anti-CD9, the probability of detecting various exosomes in a blood sample is greatly increased, thereby achieving the goal of expanding the detection range.

[0065] The three channels of the present invention adopt a consistent structural design, which can effectively control variables, so that the test results of the same sample in different types of tests have good comparability, thereby achieving more accurate comparison of different types of test results and providing more research and analysis dimensions.

[0066] The above description is only a preferred specific embodiment of the present invention, but the scope of protection of the present invention is not limited thereto. Any modifications, equivalent replacements and improvements made by any technician familiar with this technical field within the technical scope disclosed by the present invention should be included in the scope of protection of the invention.

Claims

1. A three-layer chromatography channel cancer early screening test strip based on exosome proteins, characterized by: include: drainage tablets; as well as, a first, a second, and a third chromatography channel disposed around the drainage plate; The first, second and third chromatography channels each include a sample pad, a chromatography membrane, an absorbent pad and a bottom plate; The chromatography membrane is coated with a colloidal gold pad, a detection line and a quality control line; The colloidal gold pads of the three chromatography channels are coated with three gold-labeled antibodies containing different exosomes, one of which is coated on each chromatography channel; The test line is coated with an antibody that specifically binds to the substance to be tested; The quality control lines are coated with antibodies that specifically bind to the corresponding colloidal gold particles.

2. The three-layer chromatography channel cancer early screening test strip based on exosome protein according to claim 1, characterized in that: The drainage piece is sequentially connected to the sample pad, the chromatography membrane and the water absorbent pad supported by the bottom plate.

3. The three-layer chromatography channel cancer early screening test strip based on exosome protein according to claim 1, characterized in that: The colloidal gold pad, the detection line and the quality control line are separated by a chromatography membrane.

4. The three-layer chromatography channel cancer early screening test strip based on exosome protein according to claim 3, characterized in that: The distances between the colloidal gold pad, the detection line and the quality control line, as well as their setting positions, are kept consistent in the three channels.

5. The three-layer chromatography channel cancer early screening test strip based on exosome protein according to claim 4, characterized in that: The size, thickness and water absorption properties of the absorbent materials were consistent across the three chromatography channels.

6. The three-layer chromatography channel cancer early screening test strip based on exosome protein according to claim 1, characterized in that: The gold-labeled antibody is formed by combining gold particles with exoprotein antibodies.

7. The three-layer chromatography channel cancer early screening test strip based on exosome protein according to claim 6, characterized in that: The gold-labeled antibody for the exoprotein is selected from one of gold-labeled Anti-CD63, gold-labeled Anti-CD81 and gold-labeled Anti-CD9.

8. The three-layer chromatography channel cancer early screening test strip based on exosome protein according to claim 1, characterized in that: The chromatography membrane is a nitrocellulose membrane; the bottom plate is a PVC plate.

9. The three-layer chromatography channel cancer early screening test strip based on exosome protein according to claim 1, characterized in that: The first, second and third chromatography channels are evenly connected to the drainage plate.

10. The three-layer chromatography channel cancer early screening test strip based on exosome protein according to claim 1, characterized in that: The drainage piece is provided with a rear injection port.

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