Urinary trypsinogen-2 determination kit

Abstract

The utility model relates to the field of biological detection, in particular to a urinary trypsinogen-2 determination kit, which is characterized in that a fluorescence labeled urinary trypsinogen-2 antibody is contained in a fluorescence labeled pad and can be quickly combined with urinary trypsinogen-2 in a sample; due to the design of the fluorescent labeled pad and the nitrocellulose membrane, the detection reaction is rapidly carried out, and the detection time is shortened; the sample pad, the fluorescent label pad, the compact glass fiber pad, the nitrocellulose membrane and the absorbent paper are reasonably arranged on the test strip, so that uniform flow of liquid on the test strip is ensured, and the detection efficiency is improved; a user only needs to dropwise add a sample into the sample adding opening, operation is easy and fast, complex pretreatment steps are not needed, even non-professional personnel can easily operate, and the use threshold is lowered. The problems that a traditional urinary trypsinogen-2 detection method is complex in operation and long in consumed time, needs professional technicians to carry out and limits wide clinical application of the traditional urinary trypsinogen-2 detection method are solved.

Description

Technical Field

[0001] This invention relates to the field of biological detection technology, and in particular to a urine trypsinogen-2 assay kit. Background Technology

[0002] Urinary trypsinogen-2 is a valuable biomarker for the early diagnosis of acute pancreatitis. When the pancreas is inflamed or damaged, urinary trypsinogen-2 levels rise significantly, making it a sensitive indicator for assessing pancreatic disease activity. Compared to serum amylase and lipase, urinary trypsinogen-2 not only provides more accurate diagnostic information in the early stages of the disease, but its concentration changes also better reflect the severity and prognosis of the disease. However, the measurement of urinary trypsinogen-2 faces several challenges in practical application. Traditional detection methods, such as enzyme-linked immunosorbent assay (ELISA), are complex, time-consuming, and require specialized technicians, limiting their widespread clinical application.

[0003] To address the aforementioned issues, there is an urgent need for a convenient and quick urine trypsinogen-2 assay kit. Utility Model Content

[0004] To address the problem that traditional methods for detecting urinary trypsinogen-2 are complex, time-consuming, and require specialized technicians, thus limiting their widespread clinical application, a new approach is needed.

[0005] This invention provides a urinary trypsinogen-2 assay kit, comprising an IC card and a test card. The test card includes a card shell and a test strip disposed inside the card shell. The card shell comprises a card cover and a card seat, with the test strip disposed between the card cover and the card seat. The card cover has a sample inlet and an observation port. The test strip includes a base plate and, in sequence, a sample pad, a fluorescently labeled antibody pad, a dense fiberglass pad, a nitrocellulose membrane, and absorbent paper arranged on the base plate. The sample pad is disposed below the sample inlet, and the nitrocellulose membrane is disposed below the observation port. The nitrocellulose membrane has a detection line and a control line. The fluorescently labeled antibody pad is coated with fluorescently labeled urinary trypsinogen-2 antibody and fluorescently labeled chicken IgY antibody.

[0006] Preferably, the card cover is provided with a buckle, and the card seat is provided with a card slot. The number of buckles and card slots are equal and their positions correspond. When the card cover and card seat are closed, the buckles are inserted into the card slots.

[0007] Preferably, the card holder is provided with a test strip slot for holding test strips.

[0008] Preferably, the base plate is made of polystyrene.

[0009] Preferably, the length of the sample pad is 14mm-16mm, and the length of the sample inlet is 7mm-9mm.

[0010] Preferably, the length of the nitrocellulose membrane is 24mm-26mm, and the length of the observation port is 17mm-19mm.

[0011] Preferably, the length of the fluorescent label pad is 10mm-11mm, the length of the dense fiberglass pad is 6mm-8mm, and the length of the absorbent paper is 23mm-24mm.

[0012] Preferably, the test line is coated with anti-urinary trypsinogen-2 antibody, and the control line is coated with anti-chicken IgY antibody.

[0013] Preferably, the fluorescent substance used to label urinary trypsinogen-2 antibody is one of fluorescent protein, fluorescent microspheres, quantum dots and time-resolved fluorescent microspheres.

[0014] The beneficial effects of this invention are as follows: the fluorescently labeled antibody pad contains fluorescently labeled urinary trypsinogen-2 antibody, which can rapidly bind to urinary trypsinogen-2 in the sample; the design of the fluorescently labeled antibody pad and nitrocellulose membrane allows for rapid detection, shortening the detection time; the rational arrangement of the sample pad, fluorescently labeled antibody pad, dense fiberglass pad, nitrocellulose membrane, and absorbent paper on the test strip ensures uniform liquid flow on the test strip, improving detection efficiency; users only need to add the sample to the sample inlet, making the operation simple and quick, requiring no complicated pretreatment steps, and even non-professionals can easily operate it, lowering the barrier to entry. This solves the problem that traditional urinary trypsinogen-2 detection methods are complex, time-consuming, and require professional technicians, limiting their widespread clinical application. Attached Figure Description

[0015] Figure 1 This is an exploded view of the cassette provided by this utility model.

[0016] Figure 2 This is a side view of the test strip provided by this utility model.

[0017] Figure 3 This is a cross-sectional view of the test strip provided by this utility model.

[0018] In the diagram: 1-Card housing; 11-Card cover; 111-Sample inlet; 112-Observation port; 12-Card holder; 121-Card slot; 122-Test paper slot; 2-Test paper strip; 21-Base plate; 22-Sample pad; 23-Fluorescent standard pad; 24-Dense fiberglass pad; 25-Nitrocellulose membrane; 251-Detection line; 252-Control line; 26-Absorbent paper. Detailed Implementation

[0019] The technical solutions of the embodiments of this application will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of this application, and not all embodiments. Based on the embodiments of this application, all other embodiments obtained by those of ordinary skill in the art without creative effort are within the scope of protection of this application.

[0020] Reference Figures 1-3 A urinary trypsinogen-2 assay kit includes an IC card and a test card. The test card includes a card housing 1 and a test strip 2 disposed inside the card housing 1. The card housing 1 includes a card cover 11 and a card seat 12. The test strip 2 is disposed between the card cover 11 and the card seat 12. The card cover 11 has a sample inlet 111 and an observation port 112. The test strip 2 includes a base plate 21 and, in sequence, a sample pad 22, a fluorescently labeled antibody pad 23, a dense fiberglass pad 24, a nitrocellulose membrane 25, and absorbent paper 26 arranged on the base plate 21. The sample pad 22 is disposed below the sample inlet 111, and the nitrocellulose membrane 25 is disposed below the observation port 112. The nitrocellulose membrane 25 has a detection line 251 and a control line 252. The fluorescently labeled antibody pad 23 is coated with fluorescently labeled urinary trypsinogen-2 antibody and fluorescently labeled chicken IgY antibody.

[0021] The fluorescently labeled antibody pad 23 contains fluorescently labeled urinary trypsinogen-2 antibody, which can rapidly bind to urinary trypsinogen-2 in the sample. The design of the fluorescently labeled antibody pad 23 and the nitrocellulose membrane 25 enables rapid detection, shortening the detection time. The rational arrangement of the sample pad 22, fluorescently labeled antibody pad 23, dense glass fiber pad 24, nitrocellulose membrane 25, and absorbent paper 26 on the test strip 2 ensures uniform flow of liquid on the test strip 2, improving detection efficiency and stability. Users only need to add the sample to the sample inlet 111; the operation is simple and quick, requiring no complicated pretreatment steps. Even non-professionals can easily operate it, lowering the barrier to entry. This solves the problem that traditional urinary trypsinogen-2 detection methods are complex, time-consuming, and require professional technicians, limiting their widespread clinical application.

[0022] The base plate 21 provides support and maintains the structural stability of the test strip 2; the sample pad 22 is used to absorb and pre-process the sample, removing impurities; the fluorescently labeled antibody pad 23 contains fluorescently labeled urinary trypsinogen-2 antibody, which is used to specifically bind to urinary trypsinogen-2 in the sample; the dense fiberglass pad 24 is used to filter and evenly distribute the sample, ensuring uniform flow of the sample on the test strip 2; the nitrocellulose membrane 25 can separate and enrich urinary trypsinogen-2 in the sample, concentrating urinary trypsinogen-2 in a specific area, improving the sensitivity and specificity of detection; and the absorbent paper 26 is used to absorb excess liquid and promote the flow of liquid on the test strip 2.

[0023] In some implementations, refer to Figure 1 The cover 11 is provided with a buckle, and the card seat 12 is provided with a card slot 121. The number of buckles and card slots 121 are equal and their positions correspond. When the cover 11 and the card seat 12 are closed, the buckle is inserted into the card slot 121.

[0024] The buckle and slot 121 ensure a tight fit between the cover 11 and the seat 12, improving the sealing and structural stability of the test strip 2.

[0025] In some implementations, refer to Figure 1 The card holder 12 is provided with a test strip slot 122, which is used to place the test strip 2.

[0026] The test strip slot 122 serves to fix and protect the test strip 2, preventing it from being broken during transportation and use.

[0027] In some embodiments, the base plate 21 is made of polystyrene.

[0028] Polystyrene was chosen because, firstly, it has high mechanical strength and rigidity, providing stable support for test strip 2; secondly, it has good corrosion resistance and will not react with the reagents on test strip 2, ensuring the stability of the detection process and the accuracy of the results; in addition, polystyrene has relatively low production costs, making it suitable for large-scale production and application, effectively reducing the overall cost of the kit.

[0029] In some embodiments, the sample pad 22 has a length of 14mm-16mm, and the sample inlet 111 has a length of 7mm-9mm.

[0030] The length of the sample pad 22 is set to 14mm-16mm, providing sufficient surface area to absorb the sample and ensuring that the sample can fully contact and react with other parts of the test strip 2, such as the fluorescent marker pad 23 and the dense fiberglass pad 24; the length of the sample inlet 111 is set to 7mm-9mm, ensuring that the user can accurately drop the sample onto the sample pad 22 when adding the sample, avoiding sample overflow or uneven addition.

[0031] In some embodiments, the length of the nitrocellulose membrane 25 is 24mm-26mm, and the length of the observation port 112 is 17mm-19mm.

[0032] The nitrocellulose membrane 25 is set to a length of 24mm-26mm, providing sufficient surface area to ensure that the test line 251 and control line 252 have enough space to display the results. The observation port 112 is set to a length of 17mm-19mm, ensuring that the user can clearly see the progress of the test line 251 and control line 252, facilitating detection by the fluorescence immunoassay analyzer and rapid interpretation of the test results.

[0033] In some embodiments, the length of the fluorescent label pad 23 is 10mm-11mm, the length of the dense fiberglass pad 24 is 6mm-8mm, and the length of the absorbent paper 26 is 23mm-24mm.

[0034] The length of the fluorescently labeled antibody pad 23 is set to 10mm-11mm. This ensures that the target molecules in the sample can fully bind to the fluorescently labeled urinary trypsinogen-2 antibody, and also ensures a uniform flow path of the sample on the test strip 2, avoiding uneven flow caused by excessively long or short pads, which could affect the test results. The length of the dense fiberglass pad 24 is set to 6mm-8mm, providing sufficient surface area for filtering and uniformly distributing the sample. The length of the absorbent paper 26 is set to 23mm-24mm, providing sufficient absorbency to prevent liquid backflow during the test, which could affect the accuracy of the test results.

[0035] In some implementations, the test line 251 is coated with anti-urinary trypsinogen-2 antibody, and the control line 252 is coated with anti-chicken IgY antibody.

[0036] Test line 251 indicates the presence of urinary trypsinogen-2. If urinary trypsinogen-2 is present, test line 251 will fluoresce. Control line 252 verifies whether test strip 2 is functioning correctly. If control line 252 does not fluoresce, it indicates a potential problem with test strip 2, and the test results are unreliable. When test line 251 and control line 252 show fluorescence, this fluorescence can be detected at a specific wavelength. The fluorescence signal value is read using a fluorescence immunoassay analyzer, which is equipped with a light source and photoelectric sensor. Insert the IC card into the fluorescence immunoassay analyzer, click "Read IC Card," and confirm that the IC card batch number matches the test card batch number. The light source inside the fluorescence immunoassay analyzer emits light of a specific wavelength, exciting the fluorescent substances on the detection line 251 and control line 252 to emit light. A photoelectric sensor receives the fluorescence signals reflected from the detection line 251 and control line 252 and converts them into electrical signals. Based on the intensity of the received electrical signal and the calibration information provided by the IC card, the fluorescence immunoassay analyzer calculates the fluorescence intensity ratio of the detection line 251 to the control line 252, thereby determining the presence and concentration of urinary trypsinogen-2. Chicken IgY antibody is chosen because it does not bind to mammalian Fc receptors, rheumatoid factor (RF), complement factor (CF), or commonly used proteins A, G, and L, thus reducing the possibility of false positive signals and improving detection accuracy. Using both chicken IgY and urinary trypsinogen-2 antibody on control line 252 provides a dual verification mechanism. If the sample contains urinary trypsinogen-2, both parts will show a positive result, ensuring the validity of the test even if one part fails. Furthermore, because chicken IgY is a universal capture agent and the urinary trypsinogen-2 antibody targets a specific epitope, the system's ability to capture urinary trypsinogen-2 is increased, thereby improving the overall sensitivity and specificity of the detection. In addition, the relatively independent fluorescent labeling increases the upper limit of detection.

[0037] Preferably, the fluorescent substance used to label urinary trypsinogen-2 antibody is one of fluorescent protein, fluorescent microspheres, quantum dots and time-resolved fluorescent microspheres.

[0038] For illustrative purposes and not as a limitation, when the fluorescent substance labeled by the antibody is a fluorescent protein, the method for preparing the fluorescently labeled antibody is as follows: NHS-activated biotin is dissolved in anhydrous DMF organic solvent and reacts with the amino groups on the antibody in a PBS buffer system to generate a biotin-conjugated monoclonal antibody. This is then reacted with a fluorescent protein fused with streptavidin in a PBS solution containing BSA at room temperature for 10 min to obtain fluorescent antibody-protein particles. Biotin is added to terminate the reaction, and the mixture is dialyzed three times to remove excess biotin. The pH of the PBS reaction system is 6.0–8.0, and the BSA concentration is 0.5%–5.0%. Further, the molar ratio of NHS-activated biotin to urinary trypsinogen-2 antibody is 4–40:1, and the reaction ratio of biotin-labeled antibody to streptavidin fluorescent protein is 1:2–10. Further, the urinary trypsinogen-2 fluorescently labeled antibody and the fluorescently labeled chicken IgY antibody are mixed at a concentration of 0.05 mg / mL–0.5 mg / mL and sprayed onto a fluorescently labeled antibody pad.

[0039] Although embodiments of the present invention have been shown and described, it will be understood by those skilled in the art that various changes, modifications, substitutions and alterations can be made to these embodiments without departing from the principles and spirit of the present invention, the scope of which is defined by the appended claims and their equivalents.

Claims

1. A urinary trypsinogen-2 assay kit comprising an IC card and a test card, the test card comprising a card shell and a test strip arranged inside the card shell, characterized in that: The card shell comprises a card cover and a card seat, a test strip is arranged between the card cover and the card seat, the card cover is provided with a sample adding port and an observation port, the test strip comprises a bottom plate and a sample pad, a fluorescent marker pad, a dense glass fiber pad, a nitrocellulose membrane and an absorbent paper arranged in sequence on the bottom plate, the sample pad is arranged below the sample adding port, the nitrocellulose membrane is arranged below the observation port, the nitrocellulose membrane is provided with a detection line and a quality control line, and the fluorescent marker pad is coated with a fluorescently labeled urinary trypsinogen-2 antibody and a fluorescently labeled chicken IgY antibody.

2. The urinary trypsinogen-2 assay kit according to claim 1, characterized in that The card cover is provided with buckles, the card seat is provided with card grooves, the number of the buckles and the card grooves is equal, and the positions of the buckles and the card grooves correspond to each other, and when the card cover and the card seat are closed, the buckles are inserted into the card grooves.

3. The urinary trypsinogen-2 assay kit according to claim 1, characterized in that: The card seat is provided with a test strip groove for placing the test strip.

4. The urinary trypsinogen-2 assay kit according to claim 1, characterized in that: The bottom plate is made of polystyrene.

5. The urinary trypsinogen-2 assay kit according to claim 1, characterized in that: The length of the sample pad is 14mm-16mm, and the length of the sample adding port is 7mm-9mm.

6. The urinary trypsinogen-2 assay kit according to claim 1, characterized in that: The length of the nitrocellulose membrane is 24mm-26mm, and the length of the observation port is 17mm-19mm.

7. The urinary pro-tryptase-2 assay kit according to claim 1, characterized by: The length of the fluorescent marker pad is 10mm-11mm, the length of the dense glass fiber pad is 6mm-8mm, and the length of the absorbent paper is 23mm-24mm.

8. The urinary trypsinogen-2 assay kit according to claim 1, characterized in that: The detection line is coated with an anti-urinary trypsinogen-2 antibody, and the quality control line is coated with an anti-chicken IgY antibody.

9. The urinary pro-tryptase-2 assay kit according to claim 8, characterized by: The fluorescent substance for labeling the urinary trypsinogen-2 antibody is one of a fluorescent protein, a fluorescent microsphere, a quantum dot and a time-resolved fluorescent microsphere.

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