Multi-factor cardiovascular disease combined test kit

Abstract

The utility model discloses a multi-factor combined inspection kit for cardiovascular diseases. The multi-factor combined inspection kit comprises a reagent card and a sample treatment box, a uniform mixing cavity is formed in the sample treatment box, and a reagent card slot and a sample adding hole are formed in the sample treatment box; a sample treatment liquid is preloaded in a mixing cavity of the sample treatment box; the multiple factors comprise four factors, namely cardiac troponin I, peptidin, an amino-terminal brain natriuretic peptide precursor and a D-dimer; the reagent card comprises a bottom plate, a sample pad, a detection pad and a sample suction pad. The sample treating fluid contains a fluorescence-labeled specific detection antibody aiming at the four factors and a fluorescence-labeled quality control detection antibody. The kit can be used for quantitatively detecting four indexes of cardiac troponin, peptidin, amino-terminal brain natriuretic peptide precursor and D-dimer within 15 minutes at the same time, is simple to operate and accurate in result, and is suitable for outpatient and emergency treatment of hospitals and primary medical institutions.

Description

Technical Field

[0001] The utility model relates to the technical field of biotechnology diagnosis, in particular to a multi-factor cardiovascular disease combined detection kit. Background Art

[0002] Cardiovascular disease is a leading cause of death worldwide, claiming an estimated 17.9 million lives annually. Cardiovascular disease is a category of heart and blood vessel diseases that includes coronary heart disease, cerebrovascular disease, rheumatic heart disease, and other conditions. Cardiovascular disease is a common condition that poses a serious threat to human health, characterized by high prevalence, disability, and mortality. More than four-fifths of cardiovascular disease deaths are due to heart attacks and strokes, with one-third of these deaths occurring in people under the age of 70.

[0003] Cardiac troponin I (cTnI) is a regulatory protein unique to myocardial tissue that can inhibit the binding of myosin to actin and plays an important role in myocardial contraction. A large number of studies have shown that in the process of cardiovascular disease, the level of cardiac troponin I (cTnI) varies greatly and is one of the important serum markers of myocardial cell damage.

[0004] Copeptin is a novel neurohormone expressed in the hypothalamus and neurohypophysis. Due to its structural and detection advantages, it has gradually replaced vasopressin and is considered a key marker of nonspecific stress response. In recent years, copeptin detection has gained increasing clinical attention, particularly in cardiovascular disease.

[0005] Amino-terminal pro-brain natriuretic peptide (NT-proBNP) is a polypeptide synthesized and secreted by cardiomyocytes. It is the N-terminal cleavage product of the pro-brain natriuretic peptide. Brain natriuretic peptide is an important endogenous antidiuretic hormone that can maintain fluid balance by promoting the excretion of sodium and water. NT-proBNP is mainly produced by ventricular myocytes, and its synthesis and secretion are stimulated by cardiac load. When the cardiac load increases, cardiac muscle cells synthesize and release more NT-proBNP. As an important cardiac biomarker, NT-proBNP has broad clinical application prospects. Numerous studies have shown that it plays an important role in the diagnosis, prognosis assessment, and treatment monitoring of cardiovascular diseases.

[0006] D-dimer is a specific fibrin degradation product produced by cross-linked fibrin under the action of fibrase. It is the most important laboratory indicator of thrombosis and thrombolytic activity. D-dimer is widely used in the detection of cardiovascular diseases. Studies have shown that a number of cardiovascular diseases, such as venous thromboembolism, pulmonary embolism, disseminated intravascular coagulation, myocardial infarction, aortic dissection, atrial fibrillation, and ischemic stroke, are directly correlated with serum D-dimer concentrations.

[0007] Currently, detection methods for the four markers of high-sensitive cardiac troponin I (hs-cTnI), copeptin, amino-terminal pro-brain natriuretic peptide (NT-proBNP), and D-dimer are relatively limited. Although a few companies have developed magnetic bead-based chemiluminescent immunoassays, these products are expensive, inefficient, require independent calibration and quality control for multiple indicators, and are cumbersome to operate. Some companies have developed immunochromatographic kits for combined testing of several indicators, but these combined tests can lead to interference between competitive binding of samples within the conjugate pad, compromising specificity. Utility Model Content

[0008] In order to solve the above technical problems, the purpose of the present invention is to provide a multi-factor cardiovascular disease combined detection kit, which can simultaneously perform quantitative detection of four indicators: cardiac troponin, copeptin, amino-terminal pro-brain natriuretic peptide, and D-dimer within 15 minutes. It is simple to operate and has accurate results. It is suitable for use in hospital outpatient and emergency departments and primary medical institutions.

[0009] In order to achieve the above technical objectives and the above technical effects, the present invention is implemented through the following technical solutions:

[0010] A multi-factor cardiovascular disease combined test kit comprises a reagent card and a sample processing box; the sample processing box has a mixing chamber inside, and is provided with a reagent card slot and a sample addition hole; the mixing chamber of the sample processing box is pre-filled with a sample processing solution;

[0011] The multifactors include cardiac troponin I, copeptin, amino-terminal pro-brain natriuretic peptide, and D-dimer; the reagent card includes a base plate and a sample pad, a detection pad, and a sample suction pad arranged on the base plate;

[0012] The sample processing solution contains fluorescently labeled specific detection antibodies for the four factors and fluorescently labeled quality control detection antibodies;

[0013] Four detection lines and one quality control line are distributed on the detection pad. The four detection lines are respectively coated with specific capture antibodies corresponding to the four factors, and the quality control line is coated with a hapten or antibody that can bind to the quality control detection antibody.

[0014] Furthermore, the reagent card further comprises a shell, the sample pad, the detection pad, the sample suction pad and the bottom plate are installed in the shell, and an inspection window is provided on the shell.

[0015] Furthermore, the inspection window is arranged above the four detection lines and one quality control line.

[0016] Furthermore, the four detection lines and one quality control line are parallel to each other and are arranged in sequence along the length direction of the detection pad.

[0017] Furthermore, the quality control line is close to the sample suction pad; and the four detection lines are located on the same side of the quality control line.

[0018] Furthermore, the sample pad is a glass fiber pad or a non-woven fabric pad, the detection pad is made of a nitrocellulose membrane, the sample absorption pad is made of absorbent filter paper, and the bottom plate is a PVC board.

[0019] Furthermore, the quality control line is formed by coating goat anti-mouse IgG, goat anti-chicken IgY or goat anti-rabbit IgG antibodies.

[0020] Beneficial effects of the utility model:

[0021] The utility model can realize the combined detection of multi-factor cardiovascular diseases including cardiac troponin I, copeptin, amino-terminal pro-brain natriuretic peptide and D-dimer, and can quantitatively detect the above four factors within 15 minutes with accurate results, and is suitable for use in hospital outpatient and emergency departments and primary medical institutions.

[0022] The utility model processes the sample through the sample processing liquid, and simultaneously completes the binding of the labeled antibody during the process of completing the buffer system stabilization and sample dilution of the sample detection. Compared with the antibody binding step performed on the binding pad in the prior art, this process is carried out in the liquid phase, which has more sufficient binding, more complete reaction, and less interference.

[0023] The utility model completes four-item testing through an immunochromatographic reagent card. Compared with single-item single-testing methods such as chemiluminescence, it can greatly save testing costs. At the same time, it does not require large instruments and a stable environment for testing. It is suitable for ambulances, ICUs and other occasions, and is suitable for use in hospital outpatient and emergency departments and primary medical institutions. BRIEF DESCRIPTION OF THE DRAWINGS

[0024] Figure 1This is a front view of the reagent card (housing omitted) in the multi-factor cardiovascular disease combined test kit according to an embodiment of the present invention.

[0025] Figure 2 This is a top view of the reagent card (housing omitted) in the multi-factor cardiovascular disease combined test kit according to an embodiment of the present invention.

[0026] Figure 3 This is a schematic structural diagram of a sample processing box in a multi-factor cardiovascular disease combined test kit according to an embodiment of the present invention.

[0027] Figure 4 This is a schematic structural diagram of a reagent card (including a housing) in a multi-factor cardiovascular disease combined test kit according to an embodiment of the present invention.

[0028] Description of reference numerals:

[0029] 1: Reagent card, 11: Housing, 111: Inspection window, 12: Bottom plate, 13: Sample pad, 14: Detection pad, 141: Detection line, 142: Quality control line, 15: Sample suction pad; 2: Sample processing box, 21: Sample loading hole, 22: Reagent card slot. DETAILED DESCRIPTION

[0030] The preferred embodiments of the present invention are described in detail below in conjunction with the accompanying drawings so that the advantages and features of the present invention can be more easily understood by those skilled in the art, thereby making a clearer and more precise definition of the protection scope of the present invention.

[0031] In the description of the present invention, it should be understood that the terms "front", "rear", "left", "right", "up", "down", etc., indicating directions or positional relationships, are based on the directions or positional relationships shown in the accompanying drawings, and are only for the convenience of describing the present invention and simplifying the description, rather than indicating or implying that the device or element referred to must have a specific direction, be constructed and operated in a specific direction, and therefore cannot be understood as a limitation on the present invention.

[0032] like Figures 1 to 4 The multi-factor cardiovascular disease combined test kit shown includes a reagent card 1 and a sample processing box 2; the sample processing box 2 has a mixing chamber inside, and is provided with a reagent card slot 22 and a sample addition hole 21; the mixing chamber of the sample processing box 2 is pre-filled with a sample processing solution.

[0033] The multifactors include cardiac troponin I, copeptin, amino-terminal pro-brain natriuretic peptide, and D-dimer; the reagent card 1 includes a base plate 12 and a sample pad 13, a detection pad 14, and a sample suction pad 15 arranged on the base plate 12; wherein the sample pad 13 is arranged on one side of the detection pad 14, and the sample suction pad 15 is arranged on the other side of the detection pad 14.

[0034] The sample processing solution includes the following components: a buffer system, a protein, a blocking agent, fluorescently labeled specific detection antibodies for cardiac troponin I (hs-cTnI), copeptin, amino-terminal pro-brain natriuretic peptide (NT-proBNP), and D-dimer (D-Dimer), and a fluorescently labeled quality control detection antibody. The specific detection antibodies can be polyclonal or monoclonal. The fluorescent label can be a time-resolved fluorescent microsphere label or a quantum dot fluorescent label.

[0035] When time-resolved fluorescent microspheres are used as labeling agents, the specific detection antibody or the quality control detection antibody is coupled to the time-resolved fluorescent microspheres. The time-resolved fluorescent microspheres used for coupling are polystyrene microspheres filled with a lanthanide chelate selected from europium, terbium, samarium, neodymium, or dysprosium. Preferably, the fluorescent microspheres are polystyrene microspheres filled with a europium chelate.

[0036] In addition, it should be noted that time-resolved fluorescent microspheres or quantum dot fluorescent labeling technology is an existing technology, which is known to those skilled in the art and will not be described or limited here.

[0037] The buffer system can be a phosphate buffer system, a Tris buffer system, or a Hepes buffer system; the pH of the buffer system is between 6.0 and 9.0. The protein can be bovine serum albumin, mouse IgG, or a mixture of the two.

[0038] The detection pad 14 is provided with four detection lines 141 and one quality control line 142. The four detection lines 141 and the quality control line 142 are parallel to each other and arranged sequentially along the length of the detection pad 14. The quality control line 142 is located adjacent to the sample suction pad 15; the four detection lines 141 are located on the same side of the quality control line 142. The spacing between two adjacent detection lines 141 is equal to the spacing between a quality control line 142 and the detection line 141 adjacent to the quality control line 142. The four detection lines 141 are respectively coated with specific capture antibodies corresponding to the four factors, and the quality control line 142 is coated with a hapten or antibody that can bind to the quality control detection antibody.

[0039] It should be noted that the specific capture antibodies for cardiac troponin I (hs-cTnI), copeptin, amino-terminal pro-brain natriuretic peptide (NT-proBNP), and D-dimer used to coat the test line can be either polyclonal or monoclonal. The control line can be coated with a hapten, goat anti-mouse IgG, goat anti-chicken IgY, or goat anti-rabbit IgG.

[0040] The reagent card 1 also includes a housing 11, in which the sample pad 13, detection pad 14, sample suction pad 15, and base plate 12 are mounted. The sample pad 13, detection pad 14, and sample suction pad 15 are bonded and fixed using conventional adhesives in the prior art, making the arrangement of each structural layer more reliable. An inspection window 111 is provided on the housing 11. The inspection window 111 is positioned above the four detection lines 141 and the one quality control line 142.

[0041] In addition, the housing 11 of the reagent card 1 is an end-exposed structure, so that the end of the sample pad 13 is exposed to the outside. When testing is required, the reagent card can be directly inserted into the reagent card slot 22 of the sample processing box 2 to chromatograph the sample.

[0042] The sample pad 13 is a glass fiber pad or a non-woven fabric pad, the detection pad 14 is made of a nitrocellulose membrane, the sample absorbing pad 15 is made of absorbent filter paper, and the bottom plate is a PVC board.

[0043] The method of using the test kit of the present invention is as follows:

[0044] (1) Take out the reagent card 1 stored at room temperature from the sealed bag and place it flat on the testing table;

[0045] (2) Aspirating the sample to be tested that has been equilibrated to room temperature and adding it to the sample processing box 2 pre-filled with sample processing liquid through the sample addition hole 21;

[0046] (3) Let the sample processing box 2 stand for reaction for 5 minutes;

[0047] (4) Insert the reagent card 1 into the sample processing box 2 through the reagent card slot 22. When the end of the sample pad 13 contacts the sample, the sample will be automatically aspirated for chromatography.

[0048] (5) After waiting for 10 minutes, remove the reagent card 1, insert it into the corresponding detection device with the inspection window 111 facing upward;

[0049] (6) The detection device automatically performs quantitative analysis of cardiac troponin I (hs-cTnI), copeptin, amino-terminal pro-brain natriuretic peptide (NT-proBNP), and D-dimer according to a pre-set standard curve;

[0050] (7) If the quality control line does not detect a fluorescent signal or the fluorescent signal is lower than the set value, the experiment fails and needs to be retested.

[0051] The detection principle of the reagent of the present invention will not be described or limited in detail, as it is prior art and known to those skilled in the art. Qualitative determination can be made by the color depth of the test line. The concentrations of cardiac troponin I (hs-cTnI), copeptin, amino-terminal pro-brain natriuretic peptide (NT-proBNP), and D-dimer (D-Dimer) are calculated and displayed based on a standard curve pre-set within the detection device (dry-type immunofluorescence analyzer) to achieve quantitative detection.

[0052] It will be apparent to those skilled in the art that the present invention is not limited to the details of the exemplary embodiments described above, and that the present invention can be implemented in other specific forms without departing from the spirit or essential features of the present invention. Therefore, the embodiments should be considered in all respects as illustrative and non-restrictive, and the scope of the present invention is defined by the appended claims rather than the foregoing description, and it is intended that all variations that come within the meaning and range of equivalents of the claims be embraced within the present invention.

[0053] In addition, it should be understood that although this specification is described in terms of implementation methods, not every implementation method contains only one independent technical solution. This narrative method of the specification is only for the sake of clarity. Those skilled in the art should regard the specification as a whole. The technical solutions in each embodiment can also be appropriately combined to form other implementation methods that can be understood by those skilled in the art.

Claims

1. A multi-factor cardiovascular disease combined detection kit, characterized by: The invention comprises a reagent card and a sample processing box; the sample processing box has a mixing chamber inside, and is provided with a reagent card slot and a sample addition hole; the mixing chamber of the sample processing box is pre-filled with a sample processing liquid; The multifactors include cardiac troponin I, copeptin, amino-terminal pro-brain natriuretic peptide, and D-dimer; the reagent card includes a base plate and a sample pad, a detection pad, and a sample suction pad arranged on the base plate; The sample processing solution contains fluorescently labeled specific detection antibodies for the four factors and fluorescently labeled quality control detection antibodies; Four detection lines and one quality control line are distributed on the detection pad. The four detection lines are respectively coated with specific capture antibodies corresponding to the four factors, and the quality control line is coated with a hapten or antibody that can bind to the quality control detection antibody.

2. A multi-factor cardiovascular disease combined detection kit according to claim 1, characterized in that: The reagent card further comprises a shell, the sample pad, the detection pad, the sample suction pad and the bottom plate are installed in the shell, and an inspection window is provided on the shell.

3. A multi-factor cardiovascular disease combined detection kit according to claim 2, characterized in that: The inspection window is arranged above the four detection lines and one quality control line.

4. A multi-factor cardiovascular disease combined detection kit according to claim 1, characterized in that: The four detection lines and one quality control line are parallel to each other and are arranged in sequence along the length direction of the detection pad.

5. A multi-factor cardiovascular disease combined detection kit according to claim 4, characterized in that: The quality control line is close to the sample suction pad; the four detection lines are located on the same side of the quality control line.

6. A multi-factor cardiovascular disease combined detection kit according to claim 1, characterized in that: The sample pad is a glass fiber pad or a non-woven fabric pad, the detection pad is made of a nitrocellulose membrane, the sample absorption pad is made of absorbent filter paper, and the bottom plate is a PVC board.

7. A multi-factor cardiovascular disease combined detection kit according to claim 1, characterized in that: The quality control line is formed by coating goat anti-mouse IgG, goat anti-chicken IgY or goat anti-rabbit IgG antibodies.

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