Immunochromatography kit for detecting lung cancer based on saliva

Abstract

The utility model belongs to the field of medical equipment for disease detection based on biomarkers, and particularly relates to an immunochromatography kit for lung cancer detection based on saliva, which comprises a sampling area, a binding area and an analysis area which are arranged along the dialysis direction, a plurality of corresponding monoclonal antibodies 2 are deployed in the detection areas of the analysis area, a T1 detection area and a T2 detection area are sequentially arranged in the analysis area at intervals in the dialysis direction, the monoclonal antibodies 2 are arranged in the T1 detection area at intervals in the direction perpendicular to the dialysis direction, the monoclonal antibodies 2 of the same series as the T1 detection area are arranged in the T2 detection area, and the monoclonal antibodies 2 are arranged in the T2 detection area at intervals in the direction perpendicular to the dialysis direction. Compared with a kit of a double-antibody sandwich method in the prior art, the kit has the advantages that the concentration of the same monoclonal antibody 2 deployed in the T1 detection area and the concentration of the same monoclonal antibody 2 deployed in the T2 detection area are higher than those of the same monoclonal antibody 2 deployed in the T1 detection area and the T2 detection area, and compared with a kit of a double-antibody sandwich method, the kit is quantitative and qualitative and has high sensitivity, so that the detection sensitivity of specific biomarkers to be detected is improved, and the detection result is more reliable.

Description

Technical Field

[0001] This invention belongs to the field of medical devices for disease detection based on biomarkers, specifically referring to an immunochromatographic reagent kit for detecting lung cancer based on saliva. Background Technology

[0002] Currently, lung cancer detection mainly involves lung cancer biopsy, immunological testing of blood tumor markers (such as enzyme-linked immunosorbent assay, Western blotting, colloidal gold assay, and fluorescence assay), and blood nucleic acid testing. Clinical diagnosis of lung cancer primarily relies on detecting lung cancer-related biomarkers. However, thoracentesis and venous blood sampling for lung cancer-related biomarker screening are invasive procedures that can cause trauma to patients. For example, while percutaneous biopsy avoids surgical incision, there is still a small risk of needle tract implantation, and venous blood sampling may also carry risks due to improper operation, such as infection and bleeding. Furthermore, these procedures are time-consuming and complex, no longer meeting the requirements of rapid quantitative testing in large hospitals. Using non-invasive saliva (oral mucosal exudate) screening for lung cancer-related biomarkers can provide significant convenience for clinicians and patients. Existing kits using a double-antibody sandwich method exist for detecting specific biomarkers in saliva. These kits typically include a sampling zone, a binding zone, and an analytical zone, arranged along the dialysis direction of the biomarker. Within the analytical zone, a detection zone and a control zone are sequentially located. The binding zone contains different types of monoclonal antibodies (1) labeled with fluorescent microspheres for different biomarkers. The detection zone contains monoclonal antibodies (2), which correspond to monoclonal antibodies (1). Different types of monoclonal antibodies (1 and 2) are deployed alternately along the dialysis direction. During detection, saliva is collected in the sampling zone, dialyzes to the binding zone to bind with monoclonal antibody (1), and then continues dialyzing to the analytical zone to bind with monoclonal antibody (2). The luminescence in the analytical zone is used to determine the presence of specific biomarkers related to lung cancer in the saliva, thus providing a basis for lung cancer detection. However, because the concentration of specific biomarkers in saliva is usually very low (approximately 1 / 100-1 / 1000 of that in blood), and due to its high water content and complex composition, false positives are highly likely. Currently, existing double-antibody sandwich assay kits suffer from insufficient detection accuracy and low positive-negative concordance rate. Utility Model Content

[0003] The purpose of this invention is to provide an immunochromatographic reagent kit for detecting lung cancer based on saliva, which has high detection sensitivity and reliable detection results.

[0004] The technical solutions for achieving the above objectives include the following:

[0005] An immunochromatographic kit for detecting lung cancer based on saliva includes a sampling area, a binding area, and an analysis area arranged along the dialysis direction. The sampling area has a sample pad, the binding area has a binding pad, and several monoclonal antibodies 1 are deployed on the binding pad. The analysis area has a detection area with corresponding monoclonal antibodies 2. The several monoclonal antibodies 1 are arranged at intervals perpendicular to the dialysis direction. The analysis area has two detection areas, T1 and T2, arranged at intervals along the dialysis direction. The monoclonal antibodies 2 are arranged at intervals perpendicular to the dialysis direction in the T1 detection area, and the monoclonal antibodies 2 in the T1 detection area are aligned with the corresponding monoclonal antibodies 1 on the binding pad. The T2 detection area is equipped with monoclonal antibodies 2 of the same series as those in the T1 detection area. The concentration of the same monoclonal antibody 2 deployed in the T1 and T2 detection areas is higher in the latter than in the former.

[0006] Furthermore, the sampling area is provided with a bristle cluster, which includes several bristles. The bristles are hollow capillaries, and the bottom of the bristles are connected to the sample pad.

[0007] Furthermore, the inner diameter of the bristles is 50-100 µm, and the length is 2.5 cm. Bristles that are too long will make the toothbrush difficult to control in the mouth, reducing the space for movement; bristles that are too short will have a small coverage area and be difficult to control. Choosing bristles of an appropriate length is beneficial for improving saliva collection efficiency.

[0008] Furthermore, the same monoclonal antibody 2 is deployed 4-6 mm apart in both the T1 and T2 detection zones. Choosing an appropriate spacing for the same monoclonal antibody 2 at both ends can improve detection speed without mutual interference.

[0009] Furthermore, adjacent monoclonal antibodies within the same detection area are spaced 0.1-0.3 mm apart. The spacing between adjacent monoclonal antibodies is chosen to minimize mutual interference while capturing as many dialyzed biomarkers as possible.

[0010] In the aforementioned immunochromatographic kit for lung cancer detection based on saliva, all monoclonal antibodies 1 are labeled with fluorescent microspheres. A specific biomarker in saliva binds sequentially to the corresponding monoclonal antibody 2 and monoclonal antibody 1, resulting in a double-antibody sandwich reaction. Because the monoclonal antibodies in the kit are deployed at intervals perpendicular to the osmotic direction, the single specific biomarker to be detected in saliva does not interfere with other specific biomarkers during the double-antibody sandwich reaction. The different concentrations of monoclonal antibodies set in the T1 and T2 detection zones allow for gradient detection of saliva samples. Compared with existing double-antibody sandwich kits, this invention provides both quantitative and qualitative analysis, with high sensitivity, thereby improving the detection sensitivity of the specific biomarker and making the detection results more reliable. Attached Figure Description

[0011] Figure 1 This is a schematic diagram of the deployment structure of the monoclonal antibody in the kit, as shown in the example.

[0012] Figure 2 for Figure 1 Enlarged schematic diagram of part a.

[0013] In the diagram, 1. Sampling area; 1-1. Brush tuft; 2. Binding area; 3. Analysis area; 3-1. T1 detection area; 3-2. T2 detection area; 3-3. Quality control area. Detailed Implementation

[0014] The present invention will be described in detail below with reference to the embodiments.

[0015] See Figure 1 and Figure 2 In this embodiment, CEA, NSE, CA125, and anti-hemolysin O in saliva are selected as biomarkers for detection. The saliva-based immunochromatographic kit for lung cancer detection in this embodiment includes a sampling area 1, a binding area 2, and an analysis area 3 arranged along the dialysis direction. The sampling area 1 has a sample pad, and the binding area 2 has a binding pad. On the binding pad, anti-CEA monoclonal antibody 1, anti-NSE monoclonal antibody 1, anti-CA125 monoclonal antibody 1, and anti-hemolysin O monoclonal antibody 1 are deployed at intervals perpendicular to the dialysis direction. Figure 2 As shown in Figures A1, B1, C1, and D1, analysis zone 3 has two detection zones, T1 and T2, spaced apart along the dialysis direction. Quality control zone 3-3 is located behind detection zone 3-2 (T2). Within detection zone 3-1 (T1), anti-CEA monoclonal antibody 2, anti-NSE monoclonal antibody 2, anti-CA125 monoclonal antibody 2, and anti-hemolysin O monoclonal antibody 2 are deployed at intervals perpendicular to the dialysis direction. Figure 2As shown in the diagram, the spacing between monoclonal antibody 2 in detection zone 3-1 (T1) and monoclonal antibody 1 on the binding pad is 0.2 mm. Similarly, in detection zone 3-2 (T2), A2, B2, C2, and D2 are deployed perpendicular to the dialysis direction with the same spacing. Comparing the concentrations of the same monoclonal antibody 2 deployed in detection zones 3-2 (T1 and T2), the latter is higher than the former. For example, in healthy individuals, CEA levels are less than 43 ng / mL; levels greater than 100 ng / mL strongly suggest lung cancer. (Data source: SCI papers or publicly available industry data). The standard value for anti-CEA monoclonal antibody 2 in detection zone 3-1 (T1) is set at 43 ng / mL, while the standard value in T2 is set at 100 ng / mL. The concentrations of other monoclonal antibodies 2 in the two detection zones can be set according to this principle. The spacing between the same monoclonal antibody 2 in detection zones 3-2 (T1 and T2) is 6 mm. This distance reduces dialysis time and increases detection speed without causing interference between samples. In this embodiment, sampling area 1 is equipped with a bristle cluster 1-1, which includes several nylon bristles. Each nylon bristle is a hollow capillary with an inner diameter of 50µm and a length of 2.5cm. The bottom of the nylon bristles is connected to the sample pad. During use, the bristle cluster 1-1 is inserted into the oral cavity to collect saliva. The saliva enters the sample pad through the nylon capillary, making sampling more convenient.

[0016] The kit in this embodiment utilizes capillary action to automatically draw saliva into the sample pad in sampling area 1, reducing sample quality degradation caused by improper sampling operations. Compared with current common sampling methods, it is more convenient to operate, more rational in sampling, and improves the accuracy of detection. The T1 and T2 detection areas are set with different levels of monoclonal antibody 2, allowing for gradient detection of the same biomarker in saliva. If both T1 detection area 3-1 and T2 detection area 3-2 show color during detection, the result is positive for that biomarker, indicating that the saliva sample contains a large amount of that biomarker. The monoclonal antibodies targeting the same biomarker are sequentially deployed along the dialysis direction (e.g., A1, A2, and A2 are all deployed) in a straight line, reducing interference from resistance between bands and allowing for more complete antigen-antibody binding, which is beneficial for improving detection accuracy.

Claims

1. An immunochromatographic kit for detecting lung cancer based on saliva, comprising a sampling zone, a binding zone and an analysis zone arranged in a dialysis direction, the sampling zone being provided with a sample pad, the binding zone being provided with a binding pad, and a plurality of monoclonal antibodies 1 being disposed on the binding pad, and a plurality of corresponding monoclonal antibodies 2 being disposed in a detection zone of the analysis zone, characterized in that, The several monoclonal antibodies 1 are arranged in parallel with the dialysis direction, the analysis area is sequentially and spacedly provided with two detection areas T1 and T2 along the dialysis direction, monoclonal antibodies 2 are arranged in parallel with the dialysis direction in the T1 detection area, the monoclonal antibodies 2 in the T1 detection area are matched with the corresponding monoclonal antibodies 1 on the positive pad, the T2 detection area is provided with the same series of monoclonal antibodies 2 as the T1 detection area, and the concentration of the same kind of monoclonal antibodies 2 arranged in the T1 and T2 detection areas is higher in the latter than in the former.

2. The immunochromatography kit for detecting lung cancer based on saliva according to claim 1, characterized by, The sampling area is provided with a bristle cluster, and the bristle cluster comprises several bristles.

3. The immunochromatography kit for detecting lung cancer based on saliva according to claim 2, characterized by, The bristles are hollow capillaries, and the bottom end of the bristle is connected with the sample pad.

4. The immunochromatography kit for detecting lung cancer based on saliva according to claim 2, characterized by, The inner diameter of the bristle is 50-100 µm, and the length is 2-3 cm.

5. The immunochromatographic kit for detecting lung cancer based on saliva according to claim 1, characterized by, The material of the bristle is nylon.

6. The immunochromatographic kit for detecting lung cancer based on saliva according to claim 1, characterized by, The interval between the same kind of monoclonal antibodies 2 arranged in the T1 and T2 detection areas is 4-6 mm. The interval between adjacent monoclonal antibodies 2 in the same detection area is 0.1-0.3 mm.

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