Preparation method of sample pad for whole blood detection reagent

Abstract

The invention provides a preparation method of a sample pad for a whole blood detection reagent. The preparation method comprises the following steps: preparing a pretreatment solution; coating a fiber membrane with the pretreatment liquid, and then drying the fiber membrane; and sealing the fiber membrane by using an aluminum foil bag filled with a drying agent. The method has the advantages that the blood filtering rate is high, and red blood cells can be effectively prevented from entering a fibrous membrane area to interfere a detection result; the material is tough, easy to process and low in cost.

Description

Technical Field

[0001] The present invention relates to the field of biological detection, and more particularly, to a method for preparing a sample pad for a whole blood detection reagent. Background Art

[0002] Currently, the sample pads for whole blood detection reagents used in immunochromatographic reagents mostly adopt filter blood membranes or sample pads treated with anti-human RBC (Red Blood Cell) monoclonal antibodies. Their disadvantages are as follows: The filter blood membrane has excellent effects, but its material is soft and easy to break, resulting in a high scrap rate after processing, and its cost is high; although the sample pad treated with anti-human RBC monoclonal antibody has tough material and is easy to process, the blood filtration rate of the sample pad treated with anti-human RBC monoclonal antibody is not high, and in some whole blood detection reagents with high dilution multiples, it cannot completely filter the blood, causing human red blood cells to enter the polyester fiber membrane (NC membrane, Nitrocellulose Membrane) area and interfering with the detection results.

[0003] In summary, there is a need in the art to provide a method for preparing a sample pad for a whole blood detection reagent, which can overcome the defects of the prior art. Summary of the Invention

[0004] The present invention provides a method for preparing a sample pad for a whole blood detection reagent, which can solve the problems existing in the prior art. The object of the present invention is achieved through the following technical solutions.

[0005] An embodiment of the present invention provides a method for preparing a sample pad for a whole blood detection reagent, which includes the following steps: Step 1: Prepare a pretreatment solution; wherein, the components of the pretreatment solution include lectin and ε-polylysine; Step 2: Coat the pretreatment solution on the fiber membrane, and then dry the fiber membrane; and Step 3: Seal the fiber membrane with an aluminum foil bag containing a desiccant.

[0006] According to the method for preparing a sample pad for a whole blood detection reagent provided by the above embodiment of the present invention, the lectin is a plant agglutinin.

[0007] According to the method for preparing a sample pad for a whole blood detection reagent provided by the above embodiment of the present invention, the lectin is concanavalin A or soybean agglutinin.

[0008] According to the method for preparing a sample pad for a whole blood detection reagent provided by the above embodiment of the present invention, the proportion range of concanavalin A or soybean agglutinin in the pretreatment solution is 0.1% to 0.5%.

[0009] The preparation method of the sample pad for the whole blood detection reagent provided by the above-mentioned embodiment of the present invention, wherein the proportion range of ε-polylysine in the pretreatment solution is 0.5% to 2%.

[0010] The preparation method of the sample pad for the whole blood detection reagent provided by the above-mentioned embodiment of the present invention, wherein the fiber membrane is a polyester fiber membrane.

[0011] The preparation method of the sample pad for the whole blood detection reagent provided by the above-mentioned embodiment of the present invention, wherein the components of the pretreatment solution further include tris(hydroxymethyl)aminomethane or phosphate or sodium tetraborate, polyvinylpyrrolidone-10, sodium caseinate or bovine serum albumin, surfactant S9 (Tetronic1307) or S19 (Tween-20) or S20 (Tween-80), and disodium EDTA.

[0012] The preparation method of the sample pad for the whole blood detection reagent provided by the above-mentioned embodiment of the present invention, wherein the components of the pretreatment solution include tris(hydroxymethyl)aminomethane or phosphate or sodium tetraborate with a concentration range of 0.05 mol / L to 0.20 mol / L, 0.1% to 1.0% of polyvinylpyrrolidone-10, 0.1% to 1.0% of sodium caseinate or bovine serum albumin, 0.10% to 0.50% of surfactant S9 (Tetronic1307) or S19 (Tween-20) or S20 (Tween-80), and 0.5% to 1.5% of disodium EDTA.

[0013] The preparation method of the sample pad for the whole blood detection reagent provided by the above-mentioned embodiment of the present invention, wherein the pH value of the pretreatment solution is greater than or equal to 7.0 and less than or equal to 9.0.

[0014] The advantages of the preparation method of the sample pad for the whole blood detection reagent according to the embodiment of the present invention are as follows: high blood filtration rate, which can effectively avoid red blood cells entering the fiber membrane area and interfering with the detection results; tough material, easy to process and low cost. BRIEF DESCRIPTION OF THE DRAWINGS

[0015] Other features, objects and advantages of the present invention will become more apparent by referring to the following detailed description of the non-limiting embodiments of the present invention with reference to the accompanying drawings.

[0016] Figure 1 Shows a flowchart of the preparation method of the sample pad for the whole blood detection reagent according to an embodiment of the present invention; Figure 2 Shows a performance evaluation comparison chart of ε-polylysine. DETAILED DESCRIPTION OF THE EMBODIMENTS

[0017] The following describes the specific implementation manners of the present invention in conjunction with the accompanying drawings and embodiments. Through the content recorded in this specification, those skilled in the art can clearly and completely understand the technical solution of the present invention, the technical problems solved, and the technical effects produced. It can be understood that the specific embodiments described herein are only used to explain the present invention, rather than limiting the present invention. In addition, for the convenience of description, only the parts related to the present invention are shown in the drawings.

[0018] It should be noted that the structures, ratios, sizes, etc. shown in the drawings of the specification are only used to cooperate with the content recorded in the specification for those skilled in the art to understand and read, and are not used to limit the conditions under which the present invention can be implemented. Therefore, they do not have technical essential meanings. Any modification of the structure, change of the proportional relationship, or adjustment of the size, without affecting the effects that the present invention can produce and the purposes that can be achieved, should fall within the scope covered by the technical content disclosed by the present invention.

[0019] The similar terms such as "first", "second", "the", etc. cited do not represent a quantity limitation and can represent singular or plural. The terms "comprising", "including", "having" and any variations thereof involved in the present invention are intended to cover non-exclusive inclusion; for example, a process, method, system, product or device that includes a series of steps or modules is not limited to the listed steps or units, but may further include steps or units not listed, or may further include other steps or units inherent to these processes, methods, products or devices. The similar terms such as "connected", "coupled", etc. involved in the present invention are not limited to physical or mechanical connections, but may also include direct or indirect electrical connections.

[0020] Figure 1 The flowchart of the preparation method of the sample pad for the whole blood detection reagent according to an embodiment of the present invention is shown. As Figure 1 shown, it includes the following steps: Step 1: Prepare a pretreatment solution; wherein, the components of the pretreatment solution include lectin and ε-polylysine; wherein, ε-polylysine is used to promote the adhesion of cells to the solid matrix. The higher the molecular weight of the polymer, the greater the adhesion force it provides. Therefore, it can accelerate the solidification of red blood cells on the sample pad and reduce the interference of impurities on the detection result; Step 2: Coat the pretreatment solution on the fiber membrane, and then perform a drying treatment on the fiber membrane; and Step 3: Seal the fiber membrane with an aluminum foil bag containing a desiccant.

[0021] Lectin refers to a glycoprotein or sugar-binding protein purified from various plants, invertebrates, and higher animals. It is named lectin because it can agglutinate red blood cells (including blood group substances). Commonly used plant lectins (Phytoagglutin, PHA) are usually named after the plants from which they are extracted, such as Conconvalina (ConA), Wheat germ agglutinin (WGA), Peanut agglutinin (PNA), and Soybean agglutinin (SBA), etc. Lectin is the general term for them.

[0022] ε-Polylysine is a light yellow powder with strong hygroscopicity and a slightly bitter taste. It is a linear polymer of lysine. It is not affected by pH value and is heat-stable (120 °C, 20 min). It can inhibit heat-resistant bacteria, so heat treatment can be carried out after adding it. ε-Polylysine is rich in cations, has strong electrostatic interaction with substances with anions, and has good penetration ability to biological membranes, and has excellent hemostatic performance.

[0023] According to the preparation method of the sample pad for whole blood detection reagent provided by the above one embodiment of the present invention, wherein the lectin is a plant agglutinin.

[0024] According to the preparation method of the sample pad for whole blood detection reagent provided by the above one embodiment of the present invention, wherein the lectin is Conconvalina or Soybean agglutinin.

[0025] According to the preparation method of the sample pad for whole blood detection reagent provided by the above one embodiment of the present invention, the proportion range of Conconvalina or Soybean agglutinin in the pretreatment solution is 0.1% to 0.5%.

[0026] According to the preparation method of the sample pad for whole blood detection reagent provided by the above one embodiment of the present invention, the proportion range of ε-polylysine in the pretreatment solution is 0.5% to 2%.

[0027] According to the preparation method of the sample pad for whole blood detection reagent provided by the above one embodiment of the present invention, the fiber membrane is a polyester fiber membrane (NC membrane, Nitrocellulose Membrane).

[0028] The preparation method of the sample pad for whole blood detection reagent provided by the above-mentioned one embodiment of the present invention, wherein the components of the pretreatment solution further include tris (hydroxymethyl) aminomethane (Tris), or phosphate, or sodium tetraborate, polyvinylpyrrolidone-10 (PVP-10), sodium caseinate or bovine serum albumin, surfactant S9 (Tetronic1307) or S19 (Tween-20) or S20 (Tween-80), and disodium EDTA.

[0029] The preparation method of the sample pad for whole blood detection reagent provided by the above-mentioned one embodiment of the present invention, wherein the components of the pretreatment solution include tris (hydroxymethyl) aminomethane or phosphate or sodium tetraborate with a concentration range of 0.05 mol / L to 0.20 mol / L, 0.1% to 1.0% of polyvinylpyrrolidone-10, 0.1% to 1.0% of sodium caseinate or bovine serum albumin, 0.10% to 0.50% of surfactant S9 (Tetronic1307) or S19 (Tween-20) or S20 (Tween-80), and 0.5% to 1.5% of disodium EDTA.

[0030] The preparation method of the sample pad for whole blood detection reagent provided by the above-mentioned one embodiment of the present invention, wherein the pH value of the pretreatment solution is greater than or equal to 7.0 and less than or equal to 9.0.

[0031] Detection Take 100 μL of the sample pad pretreatment solution for whole blood detection reagent and the sample pad pretreatment solution prepared with anti-human RBC monoclonal antibody respectively, add them into 900 μL of 5% red blood cell suspension, shake well and start timing, record the time when the red blood cell suspension becomes clear. The results are as follows: Table 1: Comparative test results of the time for the red blood cell suspension to become clear The above data show that the aggregation efficiency of the sample pad pretreatment solution for whole blood detection reagent is improved compared with that of the sample pad pretreatment solution prepared with anti-human RBC monoclonal antibody, and the evaluation results at different concentrations are all significantly improved by 54.4%, with a significant improvement.

[0032] Similarly, the sample pad pretreatment solution for whole blood detection reagent and the sample pad pretreatment solution prepared with anti-human RBC monoclonal antibody are respectively treated with polyester fiber membrane and dried. The dried sample pad for whole blood detection reagent, anti-human RBC monoclonal antibody sample pad, and blood filtration membrane are respectively assembled with other immunochromatographic reagent materials into reagents, and 100 μL of 5% red blood cell suspension is added respectively. Record the reagent reaction results after 15 minutes. The results are as follows: Table II: Evaluation of Prefiltration Blood Effect Table III: Cost Accounting Comparison The above results show that the sample pad for whole blood detection reagent can completely filter out red blood cells so that human red blood cells do not enter the NC membrane area to interfere with the detection results, which is significantly improved compared with the sample pad treated with anti-human RBC monoclonal antibody, and the cost accounting of the sample pad for whole blood detection reagent is 51.9% lower than that of the blood filtration membrane.

[0033] In addition, the performance of the C-reactive Protein (CRP) reagent of the sample pad using ε-polylysine and the CRP reagent of the sample pad without using ε-polylysine was evaluated and compared with the system standard measured value. The results are as follows: Table IV: Performance Evaluation of ε-Polylysine The above results combined with Figure 2 , where Figure 2 The horizontal axis in the coordinate system is the control system standard measured value, and the vertical axis is the reagent measurement value of the CRP reagent of the sample pad with / without added ε-polylysine. It can be seen that the test results of the CRP reagent of the sample pad added with ε-polylysine are closer to the standard measured value (that is, in the linear equation y = Ax + B corresponding to the test results, the coefficient X of the test results of the CRP reagent of the sample pad added with ε-polylysine is closer to 1), which proves that ε-polylysine is beneficial to improving the measurement accuracy.

[0034] Detection conclusion: The erythrocyte aggregation efficiency of the present invention is 50 - 80% higher than that of anti-human RBC monoclonal antibody. Human red blood cells will not enter the NC membrane area to interfere with the detection results. The cost is 40 - 60% lower than that of the blood filtration membrane. The material is tough, easy to process and inexpensive, and can be widely used in reagent projects requiring whole blood detection.

[0035] The advantages of the preparation method of the sample pad for whole blood detection reagent according to the embodiment of the present invention are: high blood filtration rate, which can effectively avoid red blood cells entering the fiber membrane area to interfere with the detection results; the material is tough, easy to process and inexpensive.

[0036] Although the present invention has been described and illustrated with reference to specific embodiments thereof, such description and illustration are not intended to limit the present invention. It will be readily apparent to those skilled in the art that various changes may be made, and equivalent elements may be substituted within the embodiments without departing from the scope of the present invention as defined by the claims. Differences may exist between the technical reproduction and the actual device in the present invention due to variables in the manufacturing process and so on. There may be other embodiments of the present invention that are not specifically described. The specification and the drawings should be regarded as illustrative rather than restrictive, and modifications may be made in accordance with the purpose and spirit of the present invention, all of which are within the scope of the claims. Although the methods disclosed herein have been described with reference to specific operations performed in a specific order, it should be understood that these operations may be recombined, subdivided, or arranged in a different order to form equivalent methods without departing from the teachings of the present invention. Therefore, unless specifically indicated herein, the order and grouping of operations do not limit the present invention.

Claims

1. A preparation method of a sample pad for a whole blood detection reagent, characterized in that, It includes multiple steps: Step 1: Configure the pretreatment solution; wherein, the components of the pretreatment solution include lectin and ε-polylysine; Step 2: Coat the pretreatment solution on the fiber membrane, and then perform a drying treatment on the fiber membrane; and Step 3: Seal the fiber membrane using an aluminum foil bag containing a desiccant.

2. The preparation method of the sample pad for the whole blood detection reagent according to claim 1, wherein, The lectin is a plant agglutinin.

3. The preparation method of the sample pad for the whole blood detection reagent according to claim 2, wherein, The lectin is concanavalin A or soybean agglutinin.

4. The preparation method of the sample pad for the whole blood detection reagent according to claim 3, wherein The proportion range of concanavalin A or soybean agglutinin in the pretreatment solution is 0.1% to 0.5%.

5. The preparation method of the sample pad for the whole blood detection reagent according to claim 1, wherein, The proportion range of ε-polylysine in the pretreatment solution is 0.5% to 2%.

6. The preparation method of the sample pad for the whole blood detection reagent according to claim 1, characterized in that, The fiber membrane is a polyester fiber membrane.

7. The preparation method of the sample pad for the whole blood detection reagent according to claim 1, wherein, The components of the pretreatment solution further include tris(hydroxymethyl)aminomethane or phosphate or sodium tetraborate, polyvinylpyrrolidone-10, sodium caseinate or bovine serum albumin, surfactant S9 (Tetronic1307) or S19 (Tween-20) or S20 (Tween-80), and disodium EDTA.

8. The preparation method of the sample pad for the whole blood detection reagent according to claim 7, wherein, The components of the pretreatment solution include tris(hydroxymethyl)aminomethane or phosphate or sodium tetraborate with a concentration range of 0.05 mol / L to 0.20 mol / L, 0.1% to 1.0% of polyvinylpyrrolidone-10, 0.1% to 1.0% of sodium caseinate or bovine serum albumin, 0.10% to 0.50% of surfactant S9 (Tetronic1307) or S19 (Tween-20) or S20 (Tween-80), and 0.5% to 1.5% of disodium EDTA.

9. The preparation method of the sample pad for the whole blood detection reagent according to claim 8, characterized in that, The pH value of the pretreatment solution is greater than or equal to 7.0 and less than or equal to 9.0.

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