Immunochromatographic detection kit for platelet GP IIb / IIIa receptor inhibition rate, preparation method and detection method

Abstract

The present invention relates to the field of immunochromatographic detection, and specifically relates to an immunochromatographic detection kit for the inhibition rate of platelet GP IIb / IIIa receptor, a preparation method and a detection method, including: in the test strip A and the sample diluent: the sample diluent, the sample pad and / or the conjugate pad contain fluorescently labeled platelet surface specific receptor antibodies, anti-red blood cell antibodies, thrombin receptor I activating peptide and thrombin receptor IV activating peptide; in the test strip B and the sample diluent: the sample diluent and / or the sample-conjugate integrated pad contain fluorescently labeled platelet surface specific receptor antibodies, anti-red blood cell antibodies, thrombin receptor I activating peptide and thrombin receptor IV activating peptide; the above-mentioned kit has high detection accuracy, and has been compared with the "gold standard" turbidimetry (LTA method). The comparison results show that: the detection results of the present invention are in good agreement with the "gold standard".

Description

Technical Field

[0001] The present invention relates to the field of immunochromatographic detection, and particularly relates to an immunochromatographic detection kit for platelet GP IIb / IIIa receptor inhibition rate, a preparation method and a detection method. Background Art

[0002] Platelets play a key role in maintaining normal hemostasis. When exposed to damaged blood vessels, platelets will adhere to the exposed subendothelial matrix. Subsequently, various factors (such as thrombin, adenosine diphosphate (ADP) or collagen, etc.) released or generated at the injury site activate platelets. After platelet activation, the conformation of glycoprotein GPIIb / IIIa receptors on platelets changes, enabling them to bind to adhesion proteins such as fibrinogen, resulting in the recruitment of neighboring platelets to the injury site and aggregation to form a hemostatic plug or thrombus.

[0003] Currently, the commonly used antiplatelet drugs on the market are mainly cyclooxygenase-1 inhibitors (main representative drug: aspirin), P2Y12 receptor inhibitors (representative drugs: clopidogrel, prasugrel and ticagrelor), and GP IIb / IIIa inhibitors (representative drugs: abciximab, eptifibatide, tirofiban). GP IIb / IIIa receptor inhibitors block GP IIb / IIIa receptors in a reversible or irreversible manner, thereby preventing platelets from being activated through the GP IIb / IIIa receptor pathway.

[0004] Clinically, most patients taking GP IIb / IIIa receptor inhibitors have relatively serious and urgent thromboembolic risks. Therefore, for patients taking GP IIb / IIIa receptor inhibitors, monitoring their medication effects is of great significance. In actual situations, due to the differences in individual platelet function activities and drug efficacies, this may lead to patients taking GP IIb / IIIa receptor inhibitors not achieving the ideal medication effects. Therefore, clinically detecting the GP IIb / IIIa receptor inhibition rate of patients is very necessary for guiding their GP IIb / IIIa receptor inhibitor treatment.

[0005] Currently, there is no GP IIb / IIIa receptor inhibition rate detection product on the domestic market. Abroad, only VerifyNow has reported a GP IIb / IIIa detection product, but there is no report on an immunochromatographic method for detecting platelet GP IIb / IIIa receptor inhibition rate. Summary of the Invention

[0006] Therefore, the technical problem to be solved by the present invention is to provide an immunochromatographic detection kit for platelet GP IIb / IIIa receptor inhibition rate, a preparation method and a detection method.

[0007] To this end, the present invention provides the following technical solutions:

[0008] An immunochromatographic detection kit for the inhibition rate of platelet GP IIb / IIIa receptor, comprising: a test strip and a sample diluent; or a test strip;

[0009] The test strip includes a bottom plate, and a sample pad, a conjugate pad, a chromatographic membrane and an absorbent pad are sequentially overlapped and lapped on the bottom plate, and the test strip is denoted as test strip A; or

[0010] The test strip includes a bottom plate, and a sample-conjugate integrated pad, a chromatographic membrane and an absorbent pad are sequentially overlapped and lapped on the bottom plate, and the test strip is denoted as test strip B;

[0011] In the test strip A and the sample diluent, or in the test strip A:

[0012] The sample diluent, the sample pad and / or the conjugate pad contain a fluorescently labeled platelet surface specific receptor antibody and an anti-red blood cell antibody;

[0013] The sample pad, the conjugate pad and / or the chromatographic membrane are coated with thrombin receptor I activation peptide and thrombin receptor IV activation peptide;

[0014] The chromatographic membrane is provided with spaced detection lines and a quality control line, the detection line is coated with fibrinogen, and the quality control line is coated with a polyclonal antibody;

[0015] In the test strip B and the sample diluent, or in the test strip B:

[0016] The sample diluent and / or the sample-conjugate integrated pad contain a fluorescently labeled platelet surface specific receptor antibody and an anti-red blood cell antibody;

[0017] The sample-conjugate integrated pad and / or the chromatographic membrane are coated with thrombin receptor I activation peptide and thrombin receptor IV activation peptide;

[0018] The chromatographic membrane is provided with spaced detection lines and a quality control line, the detection line is coated with fibrinogen, and the quality control line is coated with a polyclonal antibody.

[0019] Optionally, the platelet surface specific receptor antibody is a GP IIb / IIIa receptor antibody, a P-selectin receptor antibody, a prostaglandin receptor antibody, a platelet chemokine receptor antibody and / or a 5-hydroxytryptamine receptor antibody;

[0020] Optionally, the platelet surface specific receptor antibody is a GP IIb / IIIa receptor antibody, a P-selectin receptor antibody and / or a 5-hydroxytryptamine receptor antibody;

[0021] Optionally, the platelet surface-specific receptor antibody is a GP IIb / IIIa receptor antibody and / or a P-selectin receptor antibody.

[0022] Optionally, in the test strip A and the sample diluent, the sample diluent or the sample pad contains a fluorescently labeled platelet surface-specific receptor antibody;

[0023] Optionally, in the test strip A and the sample diluent, the sample diluent contains a fluorescently labeled platelet surface-specific receptor antibody;

[0024] Optionally, in the test strip B and the sample diluent, the sample diluent contains a fluorescently labeled platelet surface-specific receptor antibody;

[0025] Optionally, in the test strip A and the sample diluent, the sample pad and / or the conjugate pad contains anti-red blood cell antibodies, thrombin receptor I activating peptide, and thrombin receptor IV activating peptide;

[0026] Optionally, in the test strip B and the sample diluent, the sample-conjugate integrated pad contains anti-red blood cell antibodies, thrombin receptor I activating peptide, and thrombin receptor IV activating peptide.

[0027] Optionally, the sample diluent contains a buffer salt at a concentration of 0.01M to 0.2M with a pH of 7.4 to 7.5; a protein protectant at a concentration of 0.2 wt% to 5 wt%; an anticoagulant at a concentration of 3.2 to 3.8 wt%; a thickening agent at a concentration of 0.2 wt% to 3 wt%; a preservative at a concentration of 0.01 wt% to 0.1 wt% and / or a surfactant at a concentration of 0.01 wt% to 0.5 wt%; and the fluorescently labeled platelet surface-specific receptor antibody at a concentration of 0.01 wt% to 2 wt%;

[0028] Optionally, the buffer salt in the sample diluent is at least one of PBS, TRIS, HEPES, and boric acid;

[0029] Optionally, the protein protectant is at least one of BSA, casein, gelatin, and trehalose;

[0030] Optionally, in the sample diluent, the anticoagulant is sodium citrate;

[0031] Optionally, the thickening agent is at least one of PVP40, PEG20000, and PVA;

[0032] Optionally, the preservative is at least one of sodium azide, P300, and gentamicin;

[0033] Optionally, the surfactant is at least one of Tween 20, Triton X-100, Tween 80, surfactant S9, and surfactant S7.

[0034] A preparation method of an immunochromatographic detection kit for the platelet GP IIb / IIIa receptor inhibition rate, comprising: the preparation of the test strip and the preparation of the sample diluent; or the preparation of the test strip; the test strip includes test strip A or test strip B;

[0035] The preparation of test strip A includes:

[0036] Soaking the sample pad with the sample pad treatment solution and drying;

[0037] Soaking the conjugate pad with the conjugate pad treatment solution and drying;

[0038] Using a membrane coating solution containing fibrinogen to draw a test line on the chromatography membrane, and using a membrane coating solution containing polyclonal antibody to draw a quality control line on the chromatography membrane, and drying;

[0039] The sample diluent, the sample pad treatment solution and / or the conjugate pad treatment solution contain a fluorescently labeled platelet surface specific receptor antibody and an anti-red blood cell antibody;

[0040] The sample pad, the conjugate pad and / or the chromatography membrane are coated with thrombin receptor I activating peptide and thrombin receptor IV activating peptide;

[0041] The sample pad, the conjugate pad, the chromatography membrane and the absorbent pad are sequentially overlapped and lapped on the bottom plate; or

[0042] The preparation of test strip B includes:

[0043] Soaking the sample-conjugate integrated pad with the sample-conjugate integrated pad treatment solution and drying;

[0044] Using a membrane coating solution containing fibrinogen to draw a test line on the chromatography membrane, and using a membrane coating solution containing polyclonal antibody to draw a quality control line on the chromatography membrane, and drying;

[0045] The sample diluent and / or the sample-conjugate integrated pad treatment solution contain a fluorescently labeled platelet surface specific receptor antibody and an anti-red blood cell antibody;

[0046] The sample-conjugate integrated pad and / or the chromatography membrane have thrombin receptor I activating peptide and thrombin receptor IV activating peptide;

[0047] The sample-conjugate integrated pad, the conjugate pad, the chromatography membrane and the absorbent pad are sequentially overlapped and lapped on the bottom plate.

[0048] Optionally, when the sample diluent contains a fluorescently labeled platelet surface specific receptor antibody, the concentration of the fluorescently labeled platelet surface specific receptor antibody is 0.01 wt% to 2 wt%; and / or

[0049] When the sample pad treatment solution contains a fluorescently labeled platelet surface specific receptor antibody, the concentration of the fluorescently labeled platelet surface specific receptor antibody is 0.01 wt% to 2 wt%; and / or

[0050] When the conjugate pad treatment solution contains a fluorescently labeled platelet surface specific receptor antibody, the concentration of the fluorescently labeled platelet surface specific receptor antibody is 0.01 wt% to 2 wt%; and / or

[0051] When the sample-conjugate integrated pad treatment solution contains a fluorescently labeled platelet surface specific receptor antibody, the concentration of the fluorescently labeled platelet surface specific receptor antibody is 0.01 wt% to 2 wt%; and / or

[0052] In the test strip A and the sample diluent, the sample pad and / or the conjugate pad contain anti-red blood cell antibodies, thrombin receptor I activating peptide, and thrombin receptor IV activating peptide; and / or

[0053] In the test strip B and the sample diluent, the sample-conjugate integrated pad contains anti-red blood cell antibodies, thrombin receptor I activating peptide, and thrombin receptor IV activating peptide.

[0054] Optionally, when the sample pad treatment solution contains a fluorescently labeled platelet surface specific receptor antibody, the concentration of thrombin receptor I activating peptide contained in the sample pad treatment solution is 1 - 20 mM, the concentration of thrombin receptor IV activating peptide is 5 - 10 mM, and the concentration of anti-red blood cell antibody is 0.01 - 0.25 mg / mL;

[0055] Optionally, when the sample pad treatment solution contains a fluorescently labeled platelet surface specific receptor antibody, the sample pad treatment solution further contains a buffer salt of 0.01 - 0.5 M, pH 7.4 - 7.5; a protein protectant of 0.2 wt% - 0.5 wt%; a preservative of 0.01 wt% - 0.05 wt%; a thickener of 0.01 wt% - 0.5 wt%; and / or a surfactant of 0.01 wt% - 0.2 wt%;

[0056] Optionally, when the sample pad treatment solution does not contain a fluorescently labeled platelet surface specific receptor antibody, the concentration of thrombin receptor I activating peptide contained in the sample pad treatment solution is 1 - 20 mM, the concentration of thrombin receptor IV activating peptide is 0.001 - 1 μM, and the concentration of anti-red blood cell antibody is 0.01 - 0.5 mg / mL;

[0057] Optionally, when the sample pad treatment solution does not contain a fluorescently labeled platelet surface specific receptor antibody, the sample pad treatment solution further contains a buffer salt at 0.01 - 0.2 M, pH 7.4 - 7.5; a protein protectant at 0.2 wt% - 2 wt%; a preservative at 0.01 wt% - 0.05 wt%; a thickener at 0.01 wt% - 0.5 wt%; and / or a surfactant at 0.01 wt% - 0.5 wt%.

[0058] Optionally, the buffer salt in the sample pad treatment solution is at least one of PBS, Tris, and HEPES;

[0059] Optionally, the protein protectant in the sample pad treatment solution is at least one of BSA, casein, and gelatin;

[0060] Optionally, the preservative in the sample pad treatment solution is at least one of sodium azide, P300, and gentamicin;

[0061] Optionally, the thickener in the sample pad treatment solution is at least one of PVP - 40, NMP, PEG40000, PVA, and polylysine;

[0062] The surfactant in the sample pad treatment solution is at least one of S7, S9, Tween 20, Tween 80, and Triton;

[0063] Optionally, the concentration of the anti - red blood cell antibody contained in the conjugate pad treatment solution is 0.01 - 0.5 mg / mL;

[0064] Optionally, the conjugate pad treatment solution further contains a buffer salt at 0.01 - 0.2 M, pH 7.4 - 7.5; a saccharide at 1 wt% - 20 wt%; a thickener at 0.01 wt% - 5 wt%; and / or a surfactant at 0.01 wt% - 0.1 wt%.

[0065] Optionally, the buffer salt in the conjugate pad treatment solution is at least one of PBS, Tris, and HEPES;

[0066] Optionally, the saccharide in the conjugate pad treatment solution is at least one of trehalose, sucrose, lactose, maltose, and glucose;

[0067] Optionally, the thickener in the conjugate pad treatment solution is at least one of PVP - 40, NMP, PEG40000, PVA, and polylysine;

[0068] Optionally, the surfactant in the conjugate pad treatment solution is at least one of S7, S9, Tween 20, Tween 80, and Triton;

[0069] Optionally, in the test strip A and the sample diluent, in the membrane coating solution containing fibrinogen, the concentration of fibrinogen is 0.5 to 2.0 mg / mL, and the rate value is 0.5 to 0.8 μL / mm;

[0070] Optionally, in the test strip A and the sample diluent, in the membrane coating solution containing polyclonal antibody, the concentration of polyclonal antibody is 0.5 to 2.0 mg / mL, and the rate value is 0.5 to 1.0 μL / mm;

[0071] Optionally, in the test strip A and the sample diluent, the membrane coating solution further contains 0.01 to 0.1 M of buffer salt, pH 7.4 to 7.5; 1 v / v% to 5 v / v% of alcohol; 0.001 wt% to 0.05 wt% of protein protectant; 0.001 wt% to 0.05 wt% of surfactant; and / or 0.01 wt% to 0.05 wt% of saccharide.

[0072] Optionally, the buffer salt in the membrane coating solution is at least one of PBS, HEPES, TRIS, and boric acid;

[0073] Optionally, the alcohol in the membrane coating solution is at least one of methanol, ethanol, and propylene glycol;

[0074] Optionally, the saccharide in the membrane coating solution is at least one of trehalose, sucrose, lactose, maltose, and glucose;

[0075] Optionally, the protein protectant in the membrane coating solution is at least one of BSA, casein, and gelatin;

[0076] Optionally, the surfactant in the membrane coating solution is at least one of S7, S9, Tween 20, Tween 80, and Triton.

[0077] Optionally, when the sample-binding integrated pad treatment solution contains a fluorescently labeled platelet surface specific receptor antibody, the concentration of thrombin receptor I activating peptide is 10 - 200 mM, the concentration of thrombin receptor IV activating peptide is 50 - 100 mM, and the concentration of anti-red blood cell antibody is 0.01 - 0.5 mg / mL;

[0078] When the sample-binding integrated pad treatment solution does not contain a fluorescently labeled platelet surface specific receptor antibody, the concentration of thrombin receptor I activating peptide is 1 - 50 mM, the concentration of thrombin receptor IV activating peptide is 5 - 100 mM, and the concentration of anti-red blood cell antibody is 0.01 - 0.5 mg / mL;

[0079] Optionally, the sample-binding integrated pad treatment solution further contains a buffer salt at a concentration of 0.01 - 0.2 M, with a pH of 7.4 - 7.5; a saccharide substance at a concentration of 1 wt% - 15 wt%; a preservative at a concentration of 0.01 wt% - 0.05 wt%; a protein protectant at a concentration of 0.2 wt% - 2 wt%; a thickening agent at a concentration of 0.01 wt% - 0.5 wt%; and / or a surfactant at a concentration of 0.01 wt% - 0.5 wt%.

[0080] Optionally, the buffer salt in the sample-binding integrated pad treatment solution is at least one of PBS, HEPES, TRIS, and boric acid;

[0081] Optionally, the saccharide substance in the sample-binding integrated pad treatment solution is at least one of trehalose, sucrose, lactose, maltose, and glucose;

[0082] Optionally, the preservative in the sample-binding integrated pad treatment solution is at least one of sodium azide, P300, and gentamicin;

[0083] Optionally, the protein protectant in the sample-binding integrated pad treatment solution is at least one of BSA, casein, and gelatin;

[0084] Optionally, the thickening agent in the sample-binding integrated pad treatment solution is at least one of PVP-40, NMP, PEG40000, PVA, and polylysine;

[0085] Optionally, the surfactant in the sample-binding integrated pad treatment solution is at least one of S7, S9, Tween 20, Tween 80, and Triton;

[0086] Optionally, in the fibrinogen-containing membrane coating solution, the concentration of fibrinogen is 0.5 - 2.0 mg / mL, and the rate value is 0.5 - 0.8 μL / mm;

[0087] Optionally, in the polyclonal antibody-containing membrane coating solution, the concentration of the polyclonal antibody is 0.5 - 2.0 mg / mL, and the rate value is 0.5 - 1.0 μL / mm;

[0088] Optionally, the membrane coating solution further contains a buffer salt at a concentration of 0.01 - 0.1 M, with a pH of 7.5; an alcohol substance at a concentration of 1 v / v% - 5 v / v%; a protein protectant at a concentration of 0.01 wt% - 0.1 wt% and / or a surfactant at a concentration of 0.01 wt% - 0.05 wt%; and / or a saccharide substance at a concentration of 0.01 wt% - 0.05 wt%.

[0089] Optionally, the buffer salt in the membrane coating solution is at least one of PBS, HEPES, TRIS, and boric acid;

[0090] Optionally, the alcohol substance in the film scribing solution is at least one of methanol, ethanol, and propylene glycol;

[0091] Optionally, the saccharide substance in the film scribing solution is at least one of trehalose, sucrose, lactose, maltose, and glucose;

[0092] Optionally, the protein protectant in the film scribing solution is at least one of BSA, casein, and gelatin;

[0093] Optionally, the surfactant in the film scribing solution is at least one of S7, S9, Tween 20, Tween 80, and Triton.

[0094] An immunochromatographic detection method for the platelet GP IIb / IIIa receptor inhibition rate, including the immunochromatographic detection kit for the platelet GP IIb / IIIa receptor inhibition rate or the immunochromatographic detection kit for the platelet GP IIb / IIIa receptor inhibition rate prepared by the preparation method.

[0095] Optionally, it includes:

[0096] Quantitative detection method:

[0097] Take the test sample and add it to or not add it to the sample diluent, then drop it onto the sample pad of test strip A or the sample-binding integrated pad of test strip B. After 3 - 10 minutes, detect the fluorescence signals of the test line and the control line respectively, and calculate the platelet GP IIb / IIIa receptor inhibition rate through the following formula:

[0098] R = [1 - T / (T + C)] × 100%;

[0099] In the formula, R represents the platelet GP IIb / IIIa receptor inhibition rate;

[0100] T represents the fluorescence signal of the test line;

[0101] C represents the fluorescence signal of the control line;

[0102] Qualitative detection method:

[0103] Take the test sample and add it to or not add it to the sample diluent, then drop it onto the sample pad of test strip A or the sample-binding integrated pad of test strip B. After 3 - 10 minutes, compare the fluorescence signals of the test line and the control line, and make a judgment according to the reference color card;

[0104] Optionally, the test sample includes whole blood sample, concentrated platelets, platelet-rich plasma, diluted whole blood sample, diluted concentrated platelets, or diluted platelet-rich plasma;

[0105] Optionally, the sample to be tested contains platelet-rich plasma or diluted platelet-rich plasma.

[0106] The technical solution of the present invention has the following advantages:

[0107] 1. An immunochromatographic detection kit for the inhibition rate of platelet GP IIb / IIIa receptor provided by the present invention includes: a test strip and a sample diluent; or a test strip; the test strip includes a bottom plate and a sample pad, a conjugate pad, a chromatographic membrane and a water absorption pad that are sequentially overlapped and lapped on the bottom plate, and the test strip is denoted as test strip A; or the test strip includes a bottom plate and a sample-conjugate integrated pad, a chromatographic membrane and a water absorption pad that are sequentially overlapped and lapped on the bottom plate, and the test strip is denoted as test strip B; in the test strip A and the sample diluent, or in the test strip A: the sample diluent, the sample pad and / or the conjugate pad contain a fluorescently labeled platelet surface-specific receptor antibody and an anti-red blood cell antibody; the sample pad, the conjugate pad and / or the chromatographic membrane are coated with thrombin receptor I activation peptide and thrombin receptor IV activation peptide; the chromatographic membrane is provided with a detection line and a quality control line at intervals, the detection line is coated with fibrinogen, and the quality control line is coated with a polyclonal antibody; in the test strip B and the sample diluent, or in the test strip B: the sample diluent and / or the sample-conjugate integrated pad contain a fluorescently labeled platelet surface-specific receptor antibody and an anti-red blood cell antibody; the sample-conjugate integrated pad and / or the chromatographic membrane are coated with thrombin receptor I activation peptide and thrombin receptor IV activation peptide; the chromatographic membrane is provided with a detection line and a quality control line at intervals, the detection line is coated with fibrinogen, and the quality control line is coated with a polyclonal antibody; in the above-mentioned kit, when detecting, after adding the sample, on the sample pad, the conjugate pad and / or the chromatographic membrane, while the thrombin receptor I activation peptide and the thrombin receptor IV activation peptide diffuse with the sample, the platelets in the sample to be tested are specifically activated by the thrombin receptor I activation peptide and the thrombin receptor IV activation peptide, and the activated platelets are captured by the fibrinogen coated on the detection line, and the unactivated platelets are captured by the polyclonal antibody coated on the quality control line. Since the sample diluent or the sample pad or the conjugate pad contains a fluorescently labeled platelet surface-specific receptor antibody, the fluorescently labeled platelet surface-specific receptor antibody can fluorescently label the activated and unactivated platelets in the sample to be tested. Therefore, by collecting the fluorescent signals on the detection line and the quality control line, the detection result can be signalized, and the inhibition degree of the GP IIb / IIIa receptor of the test sample can be qualitatively or quantitatively detected according to the fluorescent signals on the detection line and the quality control line. The same applies to test strip B.

[0108] Further, in quantitative detection, the fluorescence signals of the test line and the control line are collected, and the inhibition rate is calculated by the formula R = [1 - T / (T + C)] × 100%. The higher the inhibition rate, the higher the degree of inhibition of the GP IIb / IIIa receptor in the test sample, that is, the better the efficacy of the GP IIb / IIIa receptor inhibitor in the subject; on the contrary, the lower the inhibition rate, the worse the efficacy of the GP IIb / IIIa receptor inhibitor in the subject.

[0109] In summary, the kit of the present invention has high detection accuracy. It has been compared with the "gold standard" turbidimetry (LTA method). The comparison results show that the detection results of the present invention and the "gold standard" have good consistency.

[0110] 2. An immunochromatographic detection kit for the inhibition rate of platelet GP IIb / IIIa receptor provided by the present invention, wherein the platelet surface specific receptor antibody is a P2Y12 receptor antibody, a P-selectin receptor antibody, a prostaglandin receptor antibody, a platelet chemokine receptor antibody, and / or a 5-hydroxytryptamine receptor antibody; through long-term research, the present invention particularly selects the above-mentioned platelet surface specific receptor antibodies, and uses the platelet surface specific receptor antibodies labeled with fluorescent substances to achieve fluorescent labeling of platelets, so as to ensure that both activated platelets and unactivated platelets can be fluorescently labeled, and then the detection result signals can be obtained, and then qualitative and quantitative analysis of the detection results can be realized.

[0111] Further, anti-red blood cell antibodies are also coated on the sample pad and / or the conjugate pad. The anti-red blood cell antibodies can bind to the red blood cells in the whole blood sample to be tested, intercept the red blood cells, and avoid the red blood cells chromatographing onto the chromatographic membrane to generate background signals, which affect the detection of the signals of the test line and the quality control line, and further improve the detection accuracy. The same applies to test strip B. BRIEF DESCRIPTION OF THE DRAWINGS

[0112] In order to more clearly illustrate the specific embodiments of the present invention or the technical solutions in the prior art, the following will briefly introduce the drawings required for the description of the specific embodiments or the prior art. Obviously, the following drawings are some embodiments of the present invention. For those of ordinary skill in the art, other drawings can be obtained based on these drawings without creative efforts.

[0113] Figure 1 It is a schematic structural diagram of the test strip in Example 1 of the present invention;

[0114] Figure 2 Qualitative detection color card diagram;

[0115] Figure 3 It is a relationship diagram between the aggregation rate of the LTA method and the inhibition rate detected in Example 3 in Experimental Example 1 of the present invention;

[0116] Figure 4 It is a graph showing the relationship between the aggregation rate by the LTA method in Experimental Example 1 of the present invention and the inhibition rate detected in Example 6;

[0117] Figure 5 It is a graph showing the relationship between the aggregation rate by the LTA method in Experimental Example 1 of the present invention and the inhibition rate detected in Example 9;

[0118] Figure 6 It is a graph showing the relationship between the aggregation rate by the LTA method in Experimental Example 1 of the present invention and the inhibition rate detected in Example 12;

[0119] Figure 7 It is a graph showing the relationship between the aggregation rate by the LTA method in Experimental Example 1 of the present invention and the inhibition rate detected in Example 15.

[0120] Reference numerals:

[0121] 1 - sample pad, 2 - conjugate pad, 3 - test line, 4 - control line, 5 - chromatography membrane, 6 - absorbent pad, 7 - substrate. Detailed implementation manners

[0122] The following embodiments are provided to better further understand the present invention, and are not limited to the described optimal implementation manner. They do not constitute a limitation to the content and protection scope of the present invention. Any product that is the same as or similar to the present invention obtained by anyone under the inspiration of the present invention or by combining the features of the present invention with those of other prior arts falls within the protection scope of the present invention.

[0123] For those embodiments where specific experimental steps or conditions are not indicated, the operations or conditions of the conventional experimental steps described in the literature in this field can be followed. For the reagents or instruments whose manufacturers are not indicated, they are all conventional reagent products that can be obtained through commercial purchase.

[0124] In the present invention, the Chinese name of PVP - 40 is poly N - vinylpyrrolidone; the Chinese name of BSA is bovine serum albumin; the name of surfactant S9 is TETRONIC 1307; PBS is phosphate buffered saline; TRIS is tris(hydroxymethyl)aminomethane buffer; HEPES is 4 - (2 - hydroxyethyl)piperazine - 1 - ethanesulfonic acid; PEG20000 is polyethylene glycol 20000; PVA is polyvinyl alcohol. NMP is N - methylpyrrolidone.

[0125] The anti - red blood cell antibody in the following embodiments is purchased from Phoenix Biotech Co., Ltd.;

[0126] Fluorescently labeled P2Y12 receptor antibody, fluorescently labeled P-selectin receptor antibody, fluorescently labeled prostaglandin receptor antibody, fluorescently labeled platelet chemokine receptor antibody, fluorescently labeled 5-hydroxytryptamine receptor antibody, and polyclonal antibody (sheep anti-mouse antibody) used for quality control line were purchased from Feipeng Biotechnology Co., Ltd.

[0127] Fibrinogen was purchased from Sigma;

[0128] Thrombin receptor I activating peptide and thrombin receptor IV activating peptide were purchased from Sigma.

[0129] Examples 1-9

[0130] The sample dilution formulas of Examples 1-9 are shown in the table below.

[0131] Table 1. Concentration of each component in the sample diluent formula

[0132]

[0133] Table 1. Specific composition of each component in the sample diluent formula

[0134]

[0135]

[0136] Note: P1 represents PEG20000, P2 represents PVP40, and P3 represents PVA; P1P2 contains PEG20000 and PVP40, with a mass ratio of 4:1.

[0137] The formulas of the sample pad treatment solutions in Examples 1 to 9 are shown in the table below.

[0138] Table 2 Concentration of each component in the sample pad treatment solution formula

[0139]

[0140]

[0141] Table 2. Specific ingredients of each component in the sample pad treatment solution formula

[0142]

[0143]

[0144] The formulations of the conjugate pad treatment solutions of Examples 1 to 9 are shown in the table below.

[0145] Table 3 Concentration of each component in the conjugate pad treatment solution formulation

[0146]

[0147] Continued in Table 3, specific components of each component in the formulation of the conjugate pad treatment solution

[0148]

[0149]

[0150] The formulations of the membrane scribing solutions for the detection lines in Examples 1 - 9 are shown in the following table.

[0151] Table 4, Concentrations of each component in the formulation of the membrane scribing solution for the detection line

[0152]

[0153]

[0154] Continued in Table 4, specific components of each component in the formulation of the membrane scribing solution for the detection line

[0155]

[0156] The formulations of the membrane scribing solutions for the quality control lines in Examples 1 - 9 are shown in the following table.

[0157] Table 5, Concentrations of each component in the formulation of the membrane scribing solution for the quality control line

[0158]

[0159]

[0160] Continued in Table 5, specific components of each component in the formulation of the membrane scribing solution for the quality control line

[0161]

[0162] The preparation method of the immunochromatographic detection kit for the platelet GP IIb / IIIa receptor inhibition rate in Example 1 includes the following steps:

[0163] The sample diluent is prepared according to the sample diluent formulation in the above table, and the solvent is sterile purified water.

[0164] The preparation method of the test strip includes:

[0165] Preparation of the sample pad: Prepare according to the sample pad treatment solution formulation in the above table, with the solvent being sterile purified water; then soak the entire sample pad (made of glass fiber) in the prepared sample pad treatment solution for 10 minutes, then carefully take it out and place it on a flat carrier plate, and transfer it to an electrothermal forced air drying oven, drying temperature: 65°C, drying duration: 42 hours.

[0166] Preparation of the conjugate pad: Prepare according to the conjugate pad treatment solution formula in the above table, with the solvent being sterile purified water; then soak the entire conjugate pad (made of glass fiber) in the prepared conjugate treatment solution for 10 minutes, then carefully take it out and place it on a flat carrier plate, and transfer it to an electrothermal blast drying oven, drying temperature: 65°C, drying duration: 42 hours.

[0167] Preparation of the nitrocellulose membrane (NC membrane): Prepare according to the detection line coating solution in the above table, with the solvent being sterile water; prepare according to the quality control line coating solution in the above table, with the solvent being sterile water; select the coating surface of the nitrocellulose membrane and make a mark, and evenly coat the coating solutions of the detection line and the quality control line to be coated on the membrane in parallel, and the distance between the detection line and the quality control line is 4 - 6 mm. The NC membrane climbing speed is 70 s / 4 cm. After the coating is completed, carefully take it out and place it on a flat carrier plate, and transfer it to an electrothermal blast drying oven, drying temperature: 45°C, drying duration: 72 hours.

[0168] Group board, strip cutting and assembly: Paste the sample pad (15 mm long), conjugate pad (10 mm long), NC membrane (25 mm), and absorbent pad (30 mm) in sequence (the overlapping width of the sample pad and the conjugate pad, and the conjugate pad and the NC membrane is 2 mm; the overlapping width of the NC membrane and the absorbent pad is 4 mm) on the substrate to form a large board. The detection line is about 10 mm from the near-detection-line side edge of the conjugate pad, and the quality control line is about 10 mm from the near-quality-control-line side edge of the absorbent pad. Cut the finished large board into narrow strips of 3.5 mm. The structural schematic diagram of the prepared test strip is as Figure 1 shown, and then it can be assembled on the cartridge or card sleeve of the test strip.

[0169] An immunochromatographic detection method for platelet GP IIb / IIIa receptor inhibition rate, using the above kit for detection. Take 30 μL of the sample to be tested and add it to the sample diluent, mix well for 5 minutes, then drop it onto the sample pad, time for 3 minutes, and read and report the results by the supporting equipment (fluorescent immunoassay analyzer LEPU Quant-Fluo 800).

[0170] Record the fluorescence signals of the detection line and the quality control line respectively, and calculate the platelet GP IIb / IIIa receptor inhibition rate through the following formula:

[0171] R = [1 - T / (T + C)] × 100%;

[0172] In the formula, R represents the platelet GP IIb / IIIa receptor inhibition rate;

[0173] T represents the fluorescence signal of the detection line;

[0174] C represents the fluorescence signal of the quality control line.

[0175] Qualitative detection, compare the fluorescence signals of the test line and the quality control line, and interpret according to the reference color card as shown in Figure 2 . The percentage in Figure 2 represents the platelet GP IIb / IIIa receptor inhibition rate.

[0176] Example 2

[0177] The difference between this example and Example 1 lies in the preparation of the sample pad: the sample pad is polyester, the soaking time is 5 min, the drying temperature is 45 °C, and the drying duration is 72 h.

[0178] In the preparation of the conjugate pad: the conjugate pad is polyester, the soaking time is 5 min, the drying temperature is 45 °C, and the drying duration is 12 h.

[0179] In the preparation of the NC membrane: the NC membrane crawling speed is 140 s / 4 cm, the drying temperature is 65 °C, and the drying duration is 12 h.

[0180] Grouping, strip cutting and assembly: Cut the finished large board into narrow strips of 4.2 mm.

[0181] In the detection method: Take 50 μL of the test sample and add it to the sample diluent. After mixing for 1 minute, drop it onto the sample pad, time for 5 min, and the reading and result reporting are carried out by the supporting equipment (fluorescence immunoassay analyzer LEPU Quant-Fluo 800).

[0182] Example 3

[0183] The difference between this example and Example 1 lies in the preparation of the sample pad: the sample pad is polyester, the soaking time is 7 min, the drying temperature is 50 °C, and the drying duration is 12 h.

[0184] In the preparation of the conjugate pad: the conjugate pad is polyester, the soaking time is 7 min, the drying temperature is 50 °C, and the drying duration is 12 h.

[0185] In the preparation of the NC membrane: the NC membrane crawling speed is 185 s / 4 cm, the drying temperature is 50 °C, and the drying duration is 12 h.

[0186] Grouping, strip cutting and assembly: Cut the finished large board into narrow strips of 3.9 mm.

[0187] In the detection method: Take 40 μL of the test sample and add it to the sample diluent. After mixing for 4 minutes, drop it onto the sample pad, time for 4 min, and the reading and result reporting are carried out by the supporting equipment (fluorescence immunoassay analyzer LEPU Quant-Fluo 800).

[0188] Example 4, Example 7 are different from Example 1 in the detection method: Take 100 μL of the sample to be tested and add it to the sample diluent or directly drop it into the sample addition hole (in this example, it is chosen to add it to the sample diluent first), then drop it into the sample addition hole, time for 3 minutes, and the matching equipment will read the data and report the results.

[0189] Example 5, Example 8 are different from Example 2 in the detection method: Take 140 μL of the sample to be tested and add it to the sample diluent or directly drop it into the sample addition hole (in this example, it is chosen to add it to the sample diluent first), then drop it into the sample addition hole, time for 10 minutes, and the matching equipment will read the data and report the results.

[0190] Example 6, Example 9 are different from Example 3 in the detection method: Take 120 μL of the sample to be tested and add it to the sample diluent or directly drop it into the sample addition hole (in this example, it is chosen to add it to the sample diluent first), then drop it into the sample addition hole, time for 6 minutes, and the matching equipment will read the data and report the results.

[0191] The sample diluent formulations of Examples 10 - 15 are shown in the following table.

[0192] Table 6 Concentrations of each component in the sample diluent formulation

[0193]

[0194] Table 6 continued Specific components of each component in the sample diluent formulation

[0195]

[0196]

[0197] Note: P1 represents PEG20000, P2 represents PVP40, P3 represents PVA; P1P2 represents containing PEG20000 and PVP40 with a mass ratio of 4:1.

[0198] The formulations of the sample-binding integrated pad treatment liquid in Examples 10 - 15 are shown in the following table.

[0199] Table 7 Concentrations of each component in the sample-binding integrated pad treatment liquid formulation

[0200]

[0201]

[0202] Table 7 continued Specific components of each component in the sample-binding integrated pad treatment liquid formulation

[0203]

[0204]

[0205] The formulations of the membrane coating solutions for the detection lines in Examples 10 - 15 are shown in the following table.

[0206] Table 8. Concentrations of each component in the formulation of the membrane coating solution for the detection line

[0207]

[0208] Table 8 continued. Specific components of each component in the formulation of the membrane coating solution for the detection line

[0209]

[0210]

[0211] The formulations of the membrane coating solutions for the quality control lines in Examples 10 - 15 are shown in the following table.

[0212] Table 9. Concentrations of each component in the formulation of the membrane coating solution for the quality control line

[0213]

[0214] Table 9 continued. Specific components of each component in the formulation of the membrane coating solution for the quality control line

[0215]

[0216]

[0217] The difference between Example 10 and Example 1 lies in the preparation of the sample pad: the sample pad and the conjugate pad are combined into one pad, called the sample - conjugate integrated pad. The preparation method of the sample - conjugate integrated pad is basically the same as that of the sample pad in Example 1, and the sample pad treatment solution is replaced with the sample - conjugate integrated pad treatment solution; the sample diluent, the sample - conjugate integrated pad treatment solution, and the membrane coating solution are prepared according to the corresponding formulations in the above table;

[0218] The step of preparing the conjugate pad is omitted;

[0219] Grouping, strip cutting and assembly: Stick the sample - conjugate integrated pad (25 mm in length), NC membrane (25 mm), and absorbent pad (30 mm) on the substrate in sequence (the overlapping width between the sample - conjugate integrated pad and the NC membrane is 3 mm; the overlapping width between the NC membrane and the absorbent pad is 4 mm) to form a large board. The detection line is about 10 mm from the near - detection - line side edge of the sample - conjugate integrated pad, and the quality control line is about 10 mm from the near - quality - control - line side edge of the absorbent pad. Cut the finished large board into narrow strips with a width of 4.0 mm.

[0220] Example 11 is different from Example 2 in that in the preparation of the sample pad: the sample pad and the conjugate pad are combined into one pad, called the sample-conjugate integrated pad. The preparation method of the sample-conjugate integrated pad is basically the same as that of the sample pad in Example 1, and the sample pad treatment liquid is replaced with the sample-conjugate integrated pad treatment liquid; the sample diluent, the sample-conjugate integrated pad treatment liquid, and the membrane scribing liquid are prepared according to the corresponding formulas in the above table;

[0221] The conjugate pad preparation step is omitted;

[0222] Panel making, strip cutting and assembly: the same as in Example 10.

[0223] Example 12 is different from Example 3 in that in the preparation of the sample pad: the sample pad and the conjugate pad are combined into one pad, called the sample-conjugate integrated pad. The preparation method of the sample-conjugate integrated pad is basically the same as that of the sample pad in Example 1, and the sample pad treatment liquid is replaced with the sample-conjugate integrated pad treatment liquid; the sample diluent, the sample-conjugate integrated pad treatment liquid, and the membrane scribing liquid are prepared according to the corresponding formulas in the above table;

[0224] The conjugate pad preparation step is omitted;

[0225] Panel making, strip cutting and assembly: the same as in Example 10.

[0226] Example 13 is different from Example 4 in that in the preparation of the sample pad: the sample pad and the conjugate pad are combined into one pad, called the sample-conjugate integrated pad. The preparation method of the sample-conjugate integrated pad is basically the same as that of the sample pad in Example 1, and the sample pad treatment liquid is replaced with the sample-conjugate integrated pad treatment liquid; the sample diluent, the sample-conjugate integrated pad treatment liquid, and the membrane scribing liquid are prepared according to the corresponding formulas in the above table;

[0227] The conjugate pad preparation step is omitted;

[0228] Panel making, strip cutting and assembly: the same as in Example 10.

[0229] Example 14 is different from Example 5 in that in the preparation of the sample pad: the sample pad and the conjugate pad are combined into one pad, called the sample-conjugate integrated pad. The preparation method of the sample-conjugate integrated pad is basically the same as that of the sample pad in Example 1, and the sample pad treatment liquid is replaced with the sample-conjugate integrated pad treatment liquid; the sample diluent, the sample-conjugate integrated pad treatment liquid, and the membrane scribing liquid are prepared according to the corresponding formulas in the above table;

[0230] The conjugate pad preparation step is omitted;

[0231] Panel making, strip cutting and assembly: the same as in Example 10.

[0232] Example 15 is different from Example 6 in that in the preparation of the sample pad: the sample pad and the conjugate pad are combined into one pad, called the sample-conjugate integrated pad. The preparation method of the sample-conjugate integrated pad is basically the same as that of the sample pad in Example 1, and the sample pad treatment solution is replaced with the sample-conjugate integrated pad treatment solution; the sample diluent, the sample-conjugate integrated pad treatment solution, and the membrane coating solution are prepared according to the corresponding formulas in the above table;

[0233] The step of preparing the conjugate pad is omitted;

[0234] Grouping, strip cutting and assembly: the same as in Example 10.

[0235] Experimental Example 1

[0236] The kits and detection methods in Examples 3, 6, 9, 12, and 15 of the present invention were used for comparative tests with the "gold standard" turbidimetry (LTA method). The LTA method selected the Helena AggRAM platelet aggregation analysis system (four channels). The samples to be tested were 40 cases of (sodium citrate anticoagulated venous whole blood samples). Taking the inhibition rate obtained by the present invention as the abscissa and the inhibition rate obtained by the LTA method as the ordinate, a fitting curve was made. The results of Examples 3, 6, 9, 12, and 15 are as Figures 3 - 7 shown. It can be seen from the figure in turn that R 2 = 0.9624, R 2 = 0.9437, R 2 = 0.9475, R 2 = 0.9557, R 2 = 0.9424, indicating that the present invention has good consistency with the "gold standard".

[0237] Obviously, the above examples are only examples given for clear illustration and are not limitations on the implementation manners. For those of ordinary skill in the art, other different forms of changes or modifications can be made on the basis of the above description. It is not necessary and impossible to list all the implementation manners here. And the obvious changes or modifications derived therefrom are still within the protection scope of the present invention.

Claims

1. A preparation method of an immunochromatographic detection kit for the platelet GPIIb / IIIa receptor inhibition rate, characterized in that Comprising: The preparation of the test strip and the preparation of the sample diluent; The sample diluent contains a fluorescently labeled platelet surface specific receptor antibody; The platelet surface specific receptor antibody is a P2Y12 receptor antibody; the sample diluent also contains a buffer salt of 0.01M - 0.2M at pH 7.4 - 7.5, a protein protectant of 0.2wt% - 5wt%, an anticoagulant of 3.2 - 3.8wt%, a thickener of 0.2wt% - 3wt%, a preservative of 0.01wt% - 0.1wt%, and a surfactant of 0.01wt% - 0.5wt%; The test strip includes a bottom plate and a sample pad, a conjugate pad, a chromatographic membrane, and an absorbent pad that are sequentially overlapped and lapped on the bottom plate, and the test strip is denoted as test strip A; or The test strip includes a bottom plate and a sample-conjugate integrated pad, a chromatographic membrane, and an absorbent pad that are sequentially overlapped and lapped on the bottom plate, and the test strip is denoted as test strip B; The preparation of the test strip A includes: Soaking the sample pad with a sample pad treatment solution and drying; the sample pad treatment solution contains a buffer salt of 0.01 - 0.2M at pH 7.4 - 7.5, a protein protectant of 0.2wt% - 2wt%, a preservative of 0.01wt% - 0.05wt%, a thickener of 0.01wt% - 0.5wt%, and a surfactant of 0.01wt% - 0.5wt%; Soaking the conjugate pad with a conjugate pad treatment solution and drying; the conjugate pad treatment solution contains a buffer salt of 0.01 - 0.2M at pH 7.4 - 7.5; a saccharide substance of 1wt% - 20wt%; a thickener of 0.01wt% - 5wt%; and a surfactant of 0.01wt% - 0.1wt%; The sample pad treatment solution and / or the conjugate pad treatment solution also contains an anti-red blood cell antibody; Using a fibrinogen-containing membrane coating solution to coat the chromatographic membrane to form a test line, and using a polyclonal antibody-containing membrane coating solution to coat the chromatographic membrane to form a quality control line, and drying; The sample pad, the conjugate pad, and / or the chromatographic membrane contain thrombin receptor I activating peptide and thrombin receptor IV activating peptide; Sequentially overlapping and lapping the sample pad, the conjugate pad, the chromatographic membrane, and the absorbent pad on the bottom plate; or The preparation of the test strip B includes: Soaking the sample-conjugate integrated pad with a sample-conjugate integrated pad treatment solution and drying; the sample-conjugate integrated pad treatment solution contains a buffer salt of 0.01 - 0.2M at pH 7.4 - 7.5, a saccharide substance of 1wt% - 15wt%, a preservative of 0.01wt% - 0.05wt%, a protein protectant of 0.2wt% - 2wt%, a thickener of 0.01wt% - 0.5wt%, and a surfactant of 0.01wt% - 0.5wt%; The sample-conjugate integrated pad treatment solution also contains an anti-red blood cell antibody; Using a fibrinogen-containing membrane coating solution to coat the chromatographic membrane to form a test line, and using a polyclonal antibody-containing membrane coating solution to coat the chromatographic membrane to form a quality control line, and drying; The sample-conjugate integrated pad and / or the chromatographic membrane contain thrombin receptor I activating peptide and thrombin receptor IV activating peptide; The sample-binding integrated pad, chromatographic membrane and absorbent pad are successively overlapped and lapped on the bottom plate.

2. The preparation method of the immunochromatographic detection kit for platelet GPIIb / IIIa receptor inhibition rate according to claim 1, characterized in that, The concentration of the fluorescently labeled platelet surface specific receptor antibody contained in the sample diluent is 0.01 wt% - 2 wt%; and / or In the test strip A, the sample pad and / or the binding pad contain anti-red blood cell antibody, thrombin receptor I activating peptide and thrombin receptor IV activating peptide; In the test strip B, the sample-binding integrated pad contains anti-red blood cell antibody, thrombin receptor I activating peptide and thrombin receptor IV activating peptide.

3. The preparation method of the immunochromatographic detection kit for platelet GPIIb / IIIa receptor inhibition rate according to claim 2, characterized in that The sample pad treatment solution further contains 1 - 20 mM of thrombin receptor I activating peptide, 0.001 - 1 μM of thrombin receptor IV activating peptide, and 0.01 - 0.5 mg / mL of anti-red blood cell antibody.

4. The preparation method of the immunochromatographic detection kit for platelet GPIIb / IIIa receptor inhibition rate according to claim 1, characterized in that The buffer salt in the sample pad treatment solution is at least one of PBS, Tris, and HEPES; and / or, the protein protectant in the sample pad treatment solution is at least one of BSA, casein, and gelatin; and / or, the preservative in the sample pad treatment solution is at least one of sodium azide, P300, and gentamicin; and / or, the thickening agent in the sample pad treatment solution is at least one of PVP-40, NMP, PEG40000, PVA, and polylysine; and / or, the surfactant in the sample pad treatment solution is at least one of S7, S9, Tween 20, Tween 80, and Triton.

5. The preparation method of the immunochromatographic detection kit for platelet GPIIb / IIIa receptor inhibition rate according to claim 1, characterized in that The binding pad treatment solution further contains 0.01 - 0.5 mg / mL of anti-red blood cell antibody.

6. The preparation method of the immunochromatographic detection kit for platelet GPIIb / IIIa receptor inhibition rate according to claim 1, characterized in that The buffer salt in the binding pad treatment solution is at least one of PBS, Tris, and HEPES; and / or, the saccharide in the binding pad treatment solution is at least one of trehalose, sucrose, lactose, maltose, and glucose; and / or, the thickening agent in the binding pad treatment solution is at least one of PVP-40, NMP, PEG40000, PVA, and polylysine; and / or, the surfactant in the binding pad treatment solution is at least one of S7, S9, Tween 20, Tween 80, and Triton.

7. The preparation method of the immunochromatographic detection kit for platelet GPIIb / IIIa receptor inhibition rate according to claim 1, characterized in that In the test strip A, in the fibrinogen-containing membrane-drawing solution, the concentration of fibrinogen is 0.5 - 2.0 mg / mL, and the rate value is 0.5 - 0.8 μL / mm; And / or, in the test strip A, in the membrane coating solution containing the polyclonal antibody, the concentration of the polyclonal antibody is 0.5 - 2.0 mg / mL, and the rate value is 0.5 - 1.0 μL / mm; And / or, in the test strip A, the membrane coating solution further contains 0.01 - 0.1 M of buffer salt, with a pH of 7.4 - 7.5; 1 v / v% - 5 v / v% of alcohol; 0.001 wt% - 0.05 wt% of protein protectant; 0.001 wt% - 0.05 wt% of surfactant; and / or 0.01 wt% - 0.05 wt% of saccharide.

8. The preparation method of the immunochromatographic detection kit for the platelet GPIIb / IIIa receptor inhibition rate according to claim 7, wherein The buffer salt in the membrane coating solution is at least one of PBS, HEPES, TRIS, and boric acid; And / or, the alcohol in the membrane coating solution is at least one of methanol, ethanol, and propylene glycol; And / or, the saccharide in the membrane coating solution is at least one of trehalose, sucrose, lactose, maltose, and glucose; And / or, the protein protectant in the membrane coating solution is at least one of BSA, casein, and gelatin; And / or, the surfactant in the membrane coating solution is at least one of S7, S9, Tween 20, Tween 80, and Triton.

9. The preparation method of the immunochromatographic detection kit for the platelet GPIIb / IIIa receptor inhibition rate according to claim 2, characterized in that, The sample-binding conjugate pad treatment solution further contains 1 - 50 mM of thrombin receptor I activating peptide, 5 - 100 mM of thrombin receptor IV activating peptide, and 0.01 - 0.5 mg / mL of anti-red blood cell antibody.

10. The preparation method of the immunochromatographic detection kit for the platelet GPIIb / IIIa receptor inhibition rate according to claim 1, wherein The buffer salt in the sample-binding conjugate pad treatment solution is at least one of PBS, HEPES, TRIS, and boric acid; And / or, the saccharide in the sample-binding conjugate pad treatment solution is at least one of trehalose, sucrose, lactose, maltose, and glucose; And / or, the preservative in the sample-binding conjugate pad treatment solution is at least one of sodium azide, P300, and gentamicin; And / or, the protein protectant in the sample-binding conjugate pad treatment solution is at least one of BSA, casein, and gelatin; And / or, the thickener in the sample-binding conjugate pad treatment solution is at least one of PVP-40, NMP, PEG40000, PVA, and polylysine; And / or, the surfactant in the sample-binding conjugate pad treatment solution is at least one of S7, S9, Tween 20, Tween 80, and Triton.

11. The preparation method of the immunochromatographic detection kit for the platelet GPIIb / IIIa receptor inhibition rate according to claim 1, wherein In the test strip B, in the membrane coating solution containing fibrinogen, the concentration of fibrinogen is 0.5 - 2.0 mg / mL, and the rate value is 0.5 - 0.8 μL / mm; And / or, in the test strip B, in the membrane coating solution containing polyclonal antibody, the concentration of the polyclonal antibody is 0.5 - 2.0 mg / mL, and the rate value is 0.5 - 1.0 μL / mm; And / or, in the test strip B, the membrane coating solution further contains 0.01 - 0.1 M of buffer salt, pH 7.5; 1 v / v% - 5 v / v% of alcohol; 0.01 wt% - 0.1 wt% of protein protectant; 0.01 wt% - 0.05 wt% of surfactant; and / or 0.01 wt% - 0.05 wt% of saccharide.

12. The preparation method of the immunochromatographic detection kit for platelet GPIIb / IIIa receptor inhibition rate according to claim 11, characterized in that The buffer salt in the membrane coating solution is at least one of PBS, HEPES, TRIS, and boric acid; And / or, the alcohol in the membrane coating solution is at least one of methanol, ethanol, and propylene glycol; And / or, the saccharide in the membrane coating solution is at least one of trehalose, sucrose, lactose, maltose, and glucose; And / or, the protein protectant in the membrane coating solution is at least one of BSA, casein, and gelatin; And / or, the surfactant in the membrane coating solution is at least one of S7, S9, Tween 20, Tween 80, and Triton.

13. An immunochromatographic detection kit for the inhibition rate of platelet GPIIb / IIIa receptor, characterized in that, Comprising: The test strip and the sample diluent prepared by the preparation method according to any one of claims 1 - 12.

14. An immunochromatographic detection method for the platelet GPIIb / IIIa receptor inhibition rate for non-disease diagnosis and / or treatment purposes, characterized in that, Comprising the immunochromatographic detection kit for platelet GPIIb / IIIa receptor inhibition rate prepared by using the immunochromatographic detection kit for platelet GPIIb / IIIa receptor inhibition rate according to claim 13 or the preparation method according to any one of claims 1 - 12.

15. The immunochromatographic detection method for the platelet GPIIb / IIIa receptor inhibition rate for non-disease diagnosis and / or treatment purposes according to claim 14, characterized in that, Comprising: Quantitative detection method: Take the test sample and add it to the sample diluent, then drop it onto the sample pad of test strip A or the sample - binding integrated pad of test strip B. After 3 - 10 minutes, detect the fluorescence signals of the test line and the control line respectively, and calculate the platelet GPIIb / IIIa receptor inhibition rate through the following formula: R = [1 - T / (T + C)] × 100%; In the formula, R represents the platelet GPIIb / IIIa receptor inhibition rate; T represents the fluorescence signal of the test line; C represents the fluorescence signal of the control line; Or qualitative detection method: Take the test sample and add it to the sample diluent, then drop it onto the sample pad of test strip A or the sample - binding integrated pad of test strip B. After 3 - 10 minutes, compare the fluorescence signals of the test line and the control line, and make a judgment according to the reference color card.

16. The immunochromatographic detection method for platelet GPIIb / IIIa receptor inhibition rate for non - disease diagnosis and / or treatment purposes according to claim 15, characterized in that The test sample includes whole blood sample, concentrated platelets, or platelet - rich plasma.

17. The immunochromatographic detection method for platelet GPIIb / IIIa receptor inhibition rate for non - disease diagnosis and / or treatment purposes according to claim 15, characterized in that The sample to be tested includes a diluted whole blood sample, a diluted concentrated platelet, or a diluted platelet-rich plasma.

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