Total bilirubin dry type detection and analysis assembly and preparation method thereof

Abstract

The invention discloses a total bilirubin dry-type detection and analysis assembly and a preparation method thereof.The total bilirubin dry-type detection and analysis assembly comprises an upper supporting layer, a diffusion layer, a first reagent layer, a second reagent layer, a supporting base body and a lower supporting layer which are sequentially stacked from top to bottom; the upper supporting layer and the lower supporting layer are respectively provided with a sample adding hole and a test hole, the first reagent layer is D4 hydrogel, and the second reagent layer is prepared from a modified adhesive, acid-resistant latex paint, acid, 7-(2, 3)-dihydroxypropyl theophylline, 1, 5-sodium naphthalene disulfonate, white glue, 2, 4-dihydroxypropyl-1, 3, 4-trimethyl-1, 3, 5-trimethyl-1, 3, 5-trimethyl-1, 3, 5-trimethyl-1, 3, 5-trimethyl-1, 3, 5-trimethyl-1, 3, 5-trimethyl-1, 3, the cleaning agent is prepared from 2, 6-dichloroaniline diazonium salt, a surfactant, soluble cellulose ether, an active material and a defoaming agent.

Description

Technical Field

[0001] The invention relates to the field of detection technology, and in particular to a total bilirubin dry-type detection and analysis component and a preparation method thereof. Background Art

[0002] Existing total bilirubin detection technologies can be divided into two types: wet chemical method and dry chemical method. The advantage of wet chemical method is that the test results are accurate, but the disadvantage is that the test requires professionals and the instrument is relatively expensive. The dry chemical method realizes and makes up for the shortcomings of the wet chemical method. It has the advantages of fast detection speed, easy storage of detection reagents, no need to configure any liquid reagents, low detection threshold requirements, strong adaptability to occasions, and no need for professional operation.

[0003] At present, the commonly used dry analytical reagents continue the coating technology of photosensitive film. The shielding layer, diffusion layer, reagent layer, color development layer, etc. are coated on the transparent support in sequence according to the functional layers. The concentration of the object to be tested is detected by measuring the change of reflected light. This kind of multi-layer film dry film test results are accurate and easy to operate, but this method still has the following disadvantages:

[0004] 1) The reagent layer is unevenly colored because multiple layers are applied, resulting in poor penetration and color development.

[0005] 2) The preparation of the diffusion layer requires the use of organic solvents, which have high requirements on materials and equipment, are extremely harmful to the health of operators, and pollute the environment.

[0006] 3) The reagent needs to be stored at low temperature because fish gelatin is used, and the gelling effect of the coated fish gelatin at room temperature is poor.

[0007] 4) The production process is complicated and requires multi-layer coating because it is necessary to shield the light source, disperse the sample, and prevent the color-developing layer from oxidation and discoloration.

[0008] 5) High production cost, because the material cost increases due to the need for multi-layer coating. The prepared reagent has a large test area, because this technology uses reflectance photometry (Kubelka-Munk theory) for testing, and a small test area will result in a weak light receiving signal and insufficient sensitivity. Summary of the invention

[0009] In order to solve the above problems, the present invention provides a total bilirubin dry detection and analysis component and a preparation method thereof.

[0010] The present invention adopts the following technical solutions:

[0011] A total bilirubin dry detection and analysis component comprises an upper support layer, a diffusion layer, a first reagent layer, a second reagent layer, a support matrix and a lower support layer which are stacked in sequence from top to bottom, wherein the upper support layer and the lower support layer are respectively provided with a sample addition hole and a test hole, the first reagent layer is D4 hydrogel, and the second reagent layer comprises a modified adhesive, acid-resistant latex paint, acid, 7-(2,3)-dihydroxypropylphylline, sodium 1,5-naphthalene disulfonate, white glue, 2,6-dichloroaniline diazonium salt, a surfactant, a soluble cellulose ether, an active material and a defoaming agent.

[0012] Preferably, the concentration of the D4 hydrogel is 0.2 g / ml.

[0013] Preferably, the acid is one or more of metaphosphoric acid, tartaric acid, 5-sulfosalicylic acid, citric acid, and oxalic acid; the active material is one or more of diatomaceous earth, volcanic ash, silica gel, fly ash, microcrystalline cellulose, bentonite, titanium dioxide, and cellulose powder; the surfactant is one or more of sodium dodecylbenzene sulfonate, sodium dodecyl sulfate, hexadecyltrimethylammonium bromide, dodecylethoxysulfobetaine, and tetradecanoylamidopropylhydroxypropylsulfobetaine; the defoamer is one or more of alcohol defoamers, amide defoamers, phosphate defoamers, silicone defoamers, and polyether-modified polysiloxane defoamers.

[0014] Preferably, the amount of the acid-resistant latex paint added is 1-30g per unit square meter, the amount of the acid added is 1-30g per unit square meter, the amount of 7-(2,3)-dihydroxypropylphylline added is 1-20g per unit square meter, the amount of sodium 1,5-naphthalene disulfonate added is 1-15g per unit square meter, the amount of dichloroaniline diazonium salt added is 0.05-1g per unit square meter, the amount of the active material added is 1-40g per unit square meter, the amount of the surfactant added is 0.05-5g per unit square meter, and the amount of the defoamer added is 0.1-5ml per unit square meter.

[0015] Preferably, the diffusion layer is made of a mesh with an opening rate of 500 or 600.

[0016] Preferably, the support substrate is a plastic substrate with a transmittance of more than 80% under a wavelength of 300nm-800nm. The plastic substrate includes any one of polyethylene terephthalate, polycarbonate, polypropylene, polyethylene, polybutylene terephthalate, and polyvinyl chloride, and has a thickness of 30-200μm.

[0017] Preferably, the preparation steps of the modified adhesive specifically include:

[0018] A1. Weigh acrylamide and N-vinyl pyrrolidone in a mass ratio of 7:3 respectively, place them in 100 g of solvent, stir and dissolve them until they are completely dissolved to form solution a;

[0019] A2. Weigh 1% N,N'-methylenebisacrylamide and 0.5% N,N,N',N'-tetramethylethylenediamine respectively, add them to solution a and stir them evenly to form solution b after emulsification.

[0020] A3. Add solution b into the polymerization reactor and introduce nitrogen for 10 minutes. After the nitrogen is introduced, raise the temperature to 40°C and keep the temperature constant for 30 minutes under the protection of nitrogen. Then add 0.2% potassium persulfate and sodium bisulfite, and continue to react for 10 minutes under constant temperature conditions. Then, react for 4 hours under sealed conditions to obtain the modified adhesive.

[0021] Preferably, the modified binder is added in an amount of 1-30 g per square meter.

[0022] A method for preparing a total bilirubin dry detection and analysis component comprises the following steps:

[0023] S1, weigh 20g of D4 hydrogel and place it in 100ml of alcohol solution, wherein the ratio of alcohol to water in the alcohol solution is 9:1, and wait until it is completely dissolved to obtain a first reagent solution;

[0024] S2, respectively weighing metaphosphoric acid, 5-sulfosalicylic acid, 7-(2,3)-dihydroxypropylphylline, and sodium 1,5-naphthalene disulfonate and adding them to the aqueous solution in sequence, and wait until they are completely dissolved to obtain solution c;

[0025] S3, adding active material, acid-resistant latex paint, modified adhesive, surfactant and soluble cellulose ether to solution c respectively, and stirring to obtain solution d;

[0026] S4, continue to add 2,6-dichloroaniline diazonium salt and defoamer to solution d, continue stirring to obtain a second reagent solution;

[0027] S5, coating the first reagent solution on the support substrate, and setting the coating wet film thickness, coating speed, and coating temperature. After coating is completed, transfer to an oven at 50°C-70°C for drying for 5 minutes. After drying, a first reagent layer is formed;

[0028] S6, coating the second reagent solution on the dried first reagent layer, and setting the wet film thickness, coating speed, and coating temperature of the coating. After the coating is completed, the second reagent layer is transferred to an oven at 50°C-0°C for drying for 10 minutes, and a second reagent layer with a dry film thickness of 50-200 μm is formed after drying.

[0029] S7, cut the support matrix with the first reagent layer and the second reagent layer and the diffusion layer into 10*9.5mm 2 and assemble them according to the stacking sequence of the upper support layer, the diffusion layer, the first reagent layer, the second reagent layer, the support matrix and the lower support layer to form a total bilirubin dry detection analysis component.

[0030] Preferably, both ends of the upper supporting layer and the lower supporting layer are respectively provided with snap positions and buckles for connection and cooperation.

[0031] The dry analytical reagent test method for total bilirubin is as follows:

[0032] 10ul of the serum sample to be tested is vertically added to the sample well. The sample diffuses to the first reagent layer and the second reagent layer from the diffusion layer and is incubated at about 37°C for 400S. In this process, unconjugated bilirubin is separated from the bound protein under the action of dihydroxypropylphylline. Unconjugated bilirubin and conjugated bilirubin participate in the diazonium salt reaction together to generate a purple-red azo compound at a wavelength of 540nm. The change in reflectivity is measured by reflectance spectrophotometry, and the concentration of total bilirubin is calculated by the change in reflectivity. The equation for the reaction of the corresponding substances in the total bilirubin dry analysis reagent is as follows:

[0033]

[0034] After adopting the above technical solution, the present invention has the following advantages compared with the background technology:

[0035] The invention provides a total bilirubin dry detection and analysis component and a preparation method thereof. Under acidic conditions, a modified adhesive is combined with an acid-resistant latex paint to connect active materials, and a reagent layer is formed under the joint action of a surfactant and a defoamer. The reagent layer only needs to be coated in two layers, which improves the problem that 3-4 layers are required for coating, improves the problem of poor penetration and color development, and realizes room temperature storage, simplifies the process, and saves costs. At the same time, a gauze is used as a diffusion layer, which is environmentally friendly and harmless, does not require an organic solvent during the preparation process, is environmentally friendly, and the dry analysis reagent has good stability, is easy to store and preserve, has high detection accuracy, and a wide linear range, and can achieve rapid penetration and accurate detection of samples to be analyzed containing total bilirubin. BRIEF DESCRIPTION OF THE DRAWINGS

[0036] Figure 1 This is a schematic diagram of the structure of the total bilirubin dry type detection and analysis component of the present invention;

[0037] Figure 2 A flow chart for preparing a total bilirubin dry type detection and analysis component of the present invention;

[0038] Figure 3This is a linear correlation diagram of the test results of the bilirubin dry type detection and analysis component and the biochemical analyzer method in Example 2 of the present invention;

[0039] Figure 4 This is a linear correlation diagram of the test results of the bilirubin dry detection and analysis component and the biochemical analyzer method in Example 3 of the present invention;

[0040] Figure 5 This is a linear correlation diagram of the bilirubin dry detection and analysis component and the test results of the biochemical analyzer method in Example 4 of the present invention. DETAILED DESCRIPTION

[0041] In order to make the purpose, technical solution and advantages of the present invention more clearly understood, the present invention is further described in detail below in conjunction with the accompanying drawings and embodiments. It should be understood that the specific embodiments described herein are only used to explain the present invention and are not used to limit the present invention.

[0042] Example 1

[0043] like Figure 1 As shown, the present invention discloses a total bilirubin dry detection and analysis component, comprising an upper support layer 1, a diffusion layer 2, a first reagent layer 4, a second reagent layer 3, a support matrix 5 and a lower support layer 6 which are stacked in sequence from top to bottom, wherein the upper support layer 1 and the lower support layer 6 are respectively provided with a sample addition hole a and a test hole b, the first reagent layer is D4 hydrogel, and the second reagent layer comprises a modified adhesive, an acid-resistant latex paint, an acid, 7-(2,3)-dihydroxypropylphylline, 1,5-sodium naphthalene disulfonate, white glue (methyl vinyl ether maleic anhydride copolymer), 2,6-dichloroaniline diazonium salt, a surfactant, a soluble cellulose ether, an active material and a defoaming agent.

[0044] The concentration of D4 hydrogel is 0.2g / ml. The upper support layer 1 and the lower support layer 6 are made of black PET card sleeves, and the upper support layer 1 and the lower support layer 6 are respectively provided with a clamping position 7 and a buckle 8 at both ends for connection and matching.

[0045] The acid is one or more of metaphosphoric acid, tartaric acid, 5-sulfosalicylic acid, citric acid, and oxalic acid; the active material is one or more of diatomaceous earth, volcanic ash, silica gel, fly ash, microcrystalline cellulose, bentonite, titanium dioxide, and cellulose powder; the surfactant is one or more of sodium dodecylbenzene sulfonate, sodium dodecyl sulfate, hexadecyltrimethylammonium bromide, dodecylethoxysulfobetaine, and tetradecanoylamidopropylhydroxypropylsulfobetaine; the defoamer is one or more of alcohol defoamers, amide defoamers, phosphate defoamers, silicone defoamers, and polyether-modified polysiloxane defoamers.

[0046] The amount of acid-resistant latex paint added is 1-30g per unit square meter, the amount of acid added is 1-30g per unit square meter, the amount of 7-(2,3)-dihydroxypropylphylline added is 1-20g per unit square meter, the amount of sodium 1,5-naphthalene disulfonate added is 1-15g per unit square meter, the amount of dichloroaniline diazonium salt added is 0.05-1g per unit square meter, the amount of active material added is 1-40g per unit square meter, the amount of surfactant added is 0.05-5g per unit square meter, and the amount of defoaming agent added is 0.1-5ml per unit square meter.

[0047] The diffusion layer uses a mesh with an opening rate of 500 or 600.

[0048] The support substrate is a plastic substrate having a light transmittance of more than 80% under a wavelength of 300nm-800nm, wherein the plastic substrate includes any one of polyethylene terephthalate, polycarbonate, polypropylene, polyethylene, polybutylene terephthalate, and polyvinyl chloride, and has a thickness of 30-200μm. The plastic substrate can be a transparent plastic substrate or a frosted plastic substrate.

[0049] The preparation steps of the modified adhesive specifically include:

[0050] A1. Weigh acrylamide and N-vinyl pyrrolidone in a mass ratio of 7:3 respectively, place them in 100 g of solvent, stir and dissolve them until they are completely dissolved to form solution a;

[0051] A2. Weigh 1% N,N'-methylenebisacrylamide and 0.5% N,N,N',N'-tetramethylethylenediamine respectively, add them to solution a and stir them evenly to form solution b after emulsification.

[0052] A3, add solution b into the polymerization reactor, introduce nitrogen for 10 minutes, raise the temperature to 40°C after the nitrogen is introduced, and keep the temperature constant for 30 minutes under the protection of nitrogen, then add 0.2% potassium persulfate and sodium bisulfite, and continue to react for 10 minutes under constant temperature, and then react for 4 hours under sealed conditions to obtain a modified adhesive (poly(acrylamide-N-vinyl pyrrolidone)). The amount of the modified adhesive added is 1-30g per square meter.

[0053] The dry analytical reagent test method for total bilirubin is as follows:

[0054] 10ul of the serum sample to be tested is vertically added to the sample well. The sample diffuses to the first reagent layer and the second reagent layer from the diffusion layer and is incubated at about 37°C for 400S. In this process, unconjugated bilirubin is separated from the bound protein under the action of dihydroxypropylphylline. Unconjugated bilirubin and conjugated bilirubin participate in the diazonium salt reaction together to generate a purple-red azo compound at a wavelength of 540nm. The change in reflectivity is measured by reflectance spectrophotometry, and the concentration of total bilirubin is calculated by the change in reflectivity. The equation for the reaction of the corresponding substances in the total bilirubin dry analysis reagent is as follows:

[0055]

[0056] The serum can be a Randox quality control serum sample or a serum sample filtered from whole blood.

[0057] Example 2

[0058] like Figure 2 As shown, the present invention discloses a method for preparing a total bilirubin dry detection and analysis component, comprising the following steps:

[0059] S1. Weigh 20 g of D4 hydrogel and place it in 100 ml of alcohol solution, where the ratio of alcohol to water in the alcohol solution is 9:1, and wait until it is completely dissolved to obtain a first reagent solution;

[0060] S2. According to Table 1, weigh metaphosphoric acid, 5-sulfosalicylic acid, 7-(2,3)-dihydroxypropylphylline, and sodium 1,5-naphthalene disulfonate respectively and add them to the aqueous solution in sequence until they are completely dissolved to obtain solution c;

[0061] S3. Add active material, acid-resistant latex paint, modified adhesive, surfactant and soluble cellulose ether to solution c respectively, and stir at a stirring speed of 600 r / min for 2 h to obtain solution d. When adding acid-resistant latex paint, stirring is required to prevent the latex paint from reacting with other substances in the slurry to precipitate particles. When adding soluble cellulose ether and active material, it is also necessary to add small amounts multiple times to prevent the slurry from forming internal wrapping, resulting in uneven stirring of the slurry, thereby affecting the coating effect of the slurry. In this process, the agitator used is an electric agitator with model MYP2011-150.

[0062] S4, continue to add 2,6-dichloroaniline diazonium salt and defoamer to solution d, continue stirring, the stirring speed is 400r / min, the stirring time is 1h, and the second reagent solution is obtained;

[0063] S5, take a 125um thick frosted support substrate and place it on the coating machine, press the two ends tightly, return the micrometer to zero, apply the first reagent solution on the support substrate, and set the coating wet film thickness to 80um, the coating speed to 50cm / min, the coating temperature to 26°C, and after the coating is completed, transfer it to a 50°C oven for drying, the drying time is 5min, and after the drying is completed, the first reagent layer is formed;

[0064] S6, the dried frosted support substrate is continued to be placed on the coating machine, and the second reagent solution is coated on the dried first reagent layer, and the wet film thickness of the coating is set to 280 μm, the coating speed is 50 cm / min, and the coating temperature is 27.5°C. After the coating is completed, it is transferred to an oven at 70°C for drying, and the drying time is 10 minutes. After drying, a second reagent layer with a dry film thickness of 50-200 μm is formed;

[0065] The first reagent solution and the second reagent solution were both coated using a heated blade coater model PW-TB100.

[0066] S7, cut the support matrix with the first reagent layer and the second reagent layer and the diffusion layer into 10*9.5mm 2 The cutting equipment uses a long strip cutter with a cutting specification of 294*10mm. The total bilirubin dry detection analysis component is assembled according to the stacking sequence of the upper support layer, diffusion layer, first reagent layer, second reagent layer, support matrix and lower support layer.

[0067] The upper support layer and the lower support layer adopt black PET card sleeves, and both ends of the upper support layer and the lower support layer are respectively provided with card positions and buckles for connection and matching.

[0068] The composition ratio of the second reagent solution in this embodiment is shown in Table 1:

[0069] Table 1: Composition ratio of the second reagent solution

[0070] Reagents Coating amount / m2 Metaphosphoric acid 5g 5-Sulfosalicylic acid 7.5g 7-(2,3)-Dioxyphylline 5g Sodium 1,5-naphthalene disulfonate 3g 10% white glue 10ml Sodium Lauryl Sulfate 100mg Hexadecyltrimethylammonium bromide 100ng Acid latex paint 10g 20% Binder 15g Alcohol defoamer 1ml Silicic acid gel 30g Fly ash 5g 2,6-Dichloroaniline diazonium salt 300mg

[0071] In this example, Landau quality control was used to prepare 25 serum samples of different gradient concentrations in proportion, and the correlation of multiple test samples was analyzed by the total bilirubin dry type detection and analysis component of this example and the biochemical analyzer test method. The results are as follows Figure 3 As shown, the test results of the total bilirubin dry detection and analysis component of this embodiment and the test results of the biochemical analyzer method show a good linear correlation.

[0072] Example 3

[0073] like Figure 2 As shown, the present invention discloses a method for preparing a total bilirubin dry detection and analysis component, comprising the following steps:

[0074] S1. Weigh 20 g of D4 hydrogel and place it in 100 ml of alcohol solution, where the ratio of alcohol to water in the alcohol solution is 9:1, and wait until it is completely dissolved to obtain a first reagent solution;

[0075] S2. According to Table 1, weigh metaphosphoric acid, 5-sulfosalicylic acid, 7-(2,3)-dihydroxypropylphylline, and sodium 1,5-naphthalene disulfonate respectively and add them to the aqueous solution in sequence until they are completely dissolved to obtain solution c;

[0076] S3. Add active material, acid-resistant latex paint, modified adhesive, surfactant and soluble cellulose ether to solution c respectively, and stir at a stirring speed of 700 r / min for 2 h to obtain solution d. When adding acid-resistant latex paint, stirring is required to prevent the latex paint from reacting with other substances in the slurry to precipitate particles. When adding soluble cellulose ether and active material, it is also necessary to add small amounts multiple times to prevent the slurry from forming internal wrapping, resulting in uneven stirring of the slurry, thereby affecting the coating effect of the slurry. In this process, the agitator used is an electric agitator with model MYP2011-150.

[0077] S4, continue to add 2,6-dichloroaniline diazonium salt and defoamer to solution d, continue stirring, the stirring speed is 500r / min, the stirring time is 1h, and the second reagent solution is obtained;

[0078] S5, take a 125um thick frosted support substrate and place it on the coating machine, press the two ends tightly, return the micrometer to zero, apply the first reagent solution on the support substrate, and set the coating wet film thickness to 75um, the coating speed to 40cm / min, the coating temperature to 26°C, and after the coating is completed, transfer it to a 60°C oven for drying, the drying time is 5min, and after the drying is completed, the first reagent layer is formed;

[0079] S6, the dried frosted support substrate is continued to be placed on the coating machine, and the second reagent solution is coated on the dried first reagent layer, and the wet film thickness of the coating is set to 285 μm, the coating speed is 40 cm / min, and the coating temperature is 27.5°C. After the coating is completed, it is transferred to an oven at 60°C for drying, and the drying time is 10 minutes. After drying, a second reagent layer with a dry film thickness of 50-200 μm is formed;

[0080] The first reagent solution and the second reagent solution were both coated using a heated blade coater model PW-TB100.

[0081] S7, cut the support matrix with the first reagent layer and the second reagent layer and the diffusion layer into 10*9.5mm 2The cutting equipment uses a long strip cutter with a cutting specification of 294*10mm. The total bilirubin dry detection analysis component is assembled according to the stacking sequence of the upper support layer, diffusion layer, first reagent layer, second reagent layer, support matrix and lower support layer.

[0082] The upper support layer and the lower support layer adopt black PET card sleeves, and both ends of the upper support layer and the lower support layer are respectively provided with card positions and buckles for connection and matching.

[0083] The composition ratio of the second reagent solution of this embodiment is shown in Table 2:

[0084] Table 2: Composition ratio of the second reagent solution

[0085] Reagents Coating amount / m2 Metaphosphoric acid 5g 5-Sulfosalicylic acid 8g 7-(2,3)-Dioxyphylline 5g Sodium 1,5-naphthalene disulfonate 3.5g 10% white glue 10ml Sodium Lauryl Sulfate 50mg Hexadecyltrimethylammonium bromide 100mg Acid latex paint 10g 20% Binder 10g Alcohol defoamer 1g Silicic acid gel 25g Fly ash 7g 2,6-Dichloroaniline diazonium salt 275mg

[0086] In this example, Landau quality control was used to prepare 25 serum samples of different gradient concentrations in proportion, and the correlation of multiple test samples was analyzed by the total bilirubin dry type detection and analysis component of this example and the biochemical analyzer test method. The results are as follows Figure 4 As shown, the test results of the total bilirubin dry detection and analysis component of this embodiment and the test results of the biochemical analyzer method show a good linear correlation.

[0087] Example 4

[0088] like Figure 2 As shown, the present invention discloses a method for preparing a total bilirubin dry detection and analysis component, comprising the following steps:

[0089] S1. Weigh 20 g of D4 hydrogel and place it in 100 ml of alcohol solution, where the ratio of alcohol to water in the alcohol solution is 9:1, and wait until it is completely dissolved to obtain a first reagent solution;

[0090] S2. According to Table 1, weigh metaphosphoric acid, 5-sulfosalicylic acid, 7-(2,3)-dihydroxypropylphylline, and sodium 1,5-naphthalene disulfonate respectively and add them to the aqueous solution in sequence until they are completely dissolved to obtain solution c;

[0091] S3. Add active material, acid-resistant latex paint, modified adhesive, surfactant and soluble cellulose ether to solution c respectively, and stir at a stirring speed of 500 r / min for 2 h to obtain solution d. When adding acid-resistant latex paint, stirring is required to prevent the latex paint from reacting with other substances in the slurry to precipitate particles. When adding soluble cellulose ether and active material, it is also necessary to add small amounts multiple times to prevent the slurry from forming internal wrapping, resulting in uneven stirring of the slurry, thereby affecting the coating effect of the slurry. In this process, the agitator used is an electric agitator with model MYP2011-150.

[0092] S4, continue to add 2,6-dichloroaniline diazonium salt and defoamer to solution d, continue stirring, the stirring speed is 400r / min, the stirring time is 1h, and the second reagent solution is obtained;

[0093] S5, take a 125um thick frosted support substrate and place it on the coating machine, press the two ends tightly, return the micrometer to zero, apply the first reagent solution on the support substrate, and set the coating wet film thickness to 85um, the coating speed to 50cm / min, the coating temperature to 26°C, and after the coating is completed, transfer it to a 70°C oven for drying, the drying time is 5min, and after the drying is completed, the first reagent layer is formed;

[0094] S6, the dried frosted support substrate is continued to be placed on the coating machine, and the second reagent solution is coated on the dried first reagent layer, and the wet film thickness of the coating is set to 275 μm, the coating speed is 50 cm / min, and the coating temperature is 27.5°C. After the coating is completed, it is transferred to an oven at 70°C for drying, and the drying time is 10 minutes. After drying, a second reagent layer with a dry film thickness of 50-200 μm is formed;

[0095] The first reagent solution and the second reagent solution were both coated using a heated blade coater model PW-TB100.

[0096] S7, cut the support matrix with the first reagent layer and the second reagent layer and the diffusion layer into 10*9.5mm 2 The cutting equipment uses a long strip cutter with a cutting specification of 294*10mm. The total bilirubin dry detection analysis component is assembled according to the stacking sequence of the upper support layer, diffusion layer, first reagent layer, second reagent layer, support matrix and lower support layer.

[0097] The upper support layer and the lower support layer adopt black PET card sleeves, and both ends of the upper support layer and the lower support layer are respectively provided with card positions and buckles for connection and matching.

[0098] The composition ratio of the second reagent solution of this embodiment is shown in Table 3:

[0099] Table 3: Composition ratio of the second reagent solution

[0100] Reagents Coating amount / m2 Metaphosphoric acid 4g 5-Sulfosalicylic acid 10g 7-(2,3)-Dioxyphylline 10ml Sodium 1,5-naphthalene disulfonate 7g 10% white glue 4g Sodium Lauryl Sulfate 200mg Hexadecyltrimethylammonium bromide 50mg Acid latex paint 10g 20% Binder 12g Alcohol defoamer 2ml Silicic acid gel 25g Fly ash 5g 2,6-Dichloroaniline diazonium salt 300mg

[0101] In this example, Landau quality control was used to prepare 25 serum samples of different gradient concentrations in proportion, and the correlation of multiple test samples was analyzed by the total bilirubin dry type detection and analysis component of this example and the biochemical analyzer test method. The results are as follows Figure 5 As shown, the test results of the total bilirubin dry detection and analysis component of this embodiment and the test results of the biochemical analyzer method show a good linear correlation.

[0102] The above is only a preferred specific embodiment of the present invention, but the protection scope of the present invention is not limited thereto. Any changes or substitutions that can be easily thought of by a person skilled in the art within the technical scope disclosed by the present invention should be included in the protection scope of the present invention. Therefore, the protection scope of the present invention should be based on the protection scope of the claims.

Claims

1. A total bilirubin dry detection and analysis component, characterized in that: The invention comprises an upper support layer, a diffusion layer, a first reagent layer, a second reagent layer, a support matrix and a lower support layer which are stacked in sequence from top to bottom. The upper support layer and the lower support layer are respectively provided with a sample addition hole and a test hole. The first reagent layer is D4 hydrogel, and the second reagent layer comprises a modified adhesive, acid-resistant latex paint, acid, 7-(2,3)-dihydroxypropylphylline, sodium 1,5-naphthalene disulfonate, white glue, 2,6-dichloroaniline diazonium salt, a surfactant, a soluble cellulose ether, an active material and a defoaming agent.

2. A total bilirubin dry type detection and analysis component as claimed in claim 1, characterized in that: The concentration of the D4 hydrogel is 0.2 g / ml.

3. A total bilirubin dry type detection and analysis component as claimed in claim 1, characterized in that: The acid is one or more of metaphosphoric acid, tartaric acid, 5-sulfosalicylic acid, citric acid, and oxalic acid; the active material is one or more of diatomaceous earth, volcanic ash, silica gel, fly ash, microcrystalline cellulose, bentonite, titanium dioxide, and cellulose powder; the surfactant is one or more of sodium dodecylbenzene sulfonate, sodium dodecyl sulfate, hexadecyltrimethylammonium bromide, dodecylethoxysulfobetaine, and tetradecanoylamidopropyl hydroxypropylsulfobetaine; and the defoamer is one or more of alcohol defoamers, amide defoamers, phosphate defoamers, silicone defoamers, and polyether-modified polysiloxane defoamers.

4. A total bilirubin dry type detection and analysis component as claimed in claim 1, characterized in that: The amount of the acid-resistant latex paint added is 1-30g per unit square meter, the amount of the acid added is 1-30g per unit square meter, the amount of 7-(2,3)-dihydroxypropyltheophylline added is 1-20g per unit square meter, the amount of sodium 1,5-naphthalene disulfonate added is 1-15g per unit square meter, the amount of dichloroaniline diazonium salt added is 0.05-1g per unit square meter, the amount of the active material added is 1-40g per unit square meter, the amount of the surfactant added is 0.05-5g per unit square meter, and the amount of the defoamer added is 0.1-5ml per unit square meter.

5. A total bilirubin dry type detection and analysis component as claimed in claim 1, characterized in that: The diffusion layer is made of a mesh with an opening rate of 500 or 600.

6. A total bilirubin dry type detection and analysis component as claimed in claim 1, characterized in that: The support substrate is a plastic substrate with a light transmittance of more than 80% under a wavelength of 300nm-800nm. The plastic substrate includes any one of polyethylene terephthalate, polycarbonate, polypropylene, polyethylene, polybutylene terephthalate, and polyvinyl chloride, and has a thickness of 30-200μm.

7. A total bilirubin dry type detection and analysis component as claimed in claim 1, characterized in that: The preparation steps of the modified adhesive specifically include: A1. Weigh acrylamide and N-vinyl pyrrolidone in a mass ratio of 7:3 respectively, place them in 100 g of solvent, stir and dissolve them until they are completely dissolved to form solution a; A2. Weigh 1% N,N'-methylenebisacrylamide and 0.5% N,N,N',N'-tetramethylethylenediamine respectively, add them to solution a and stir them evenly to form solution b after emulsification. A3. Add solution b into the polymerization reactor and introduce nitrogen for 10 minutes. After the nitrogen is introduced, raise the temperature to 40°C and keep the temperature constant for 30 minutes under the protection of nitrogen. Then add 0.2% potassium persulfate and sodium bisulfite, and continue to react for 10 minutes under constant temperature conditions. Then, react for 4 hours under sealed conditions to obtain the modified adhesive.

8. A total bilirubin dry type detection and analysis component as claimed in claim 7, characterized in that: The modified binder is added in an amount of 1-30 g per square meter.

9. A method for preparing the total bilirubin dry detection and analysis component according to claims 1-8, characterized in that: The following steps are involved: S1, weigh 20g of D4 hydrogel and place it in 100ml of alcohol solution, wherein the ratio of alcohol to water in the alcohol solution is 9:1, and wait until it is completely dissolved to obtain a first reagent solution; S2, respectively weighing metaphosphoric acid, 5-sulfosalicylic acid, 7-(2,3)-dihydroxypropylphylline, and sodium 1,5-naphthalene disulfonate and adding them to the aqueous solution in sequence, and wait until they are completely dissolved to obtain solution c; S3, adding active material, acid-resistant latex paint, modified adhesive, surfactant and soluble cellulose ether to solution c respectively, and stirring to obtain solution d; S4, continue to add 2,6-dichloroaniline diazonium salt and defoamer to solution d, continue stirring to obtain a second reagent solution; S5, coating the first reagent solution on the support substrate, and setting the coating wet film thickness, coating speed, and coating temperature. After coating is completed, transfer to an oven at 50°C-70°C for drying for 5 minutes. After drying, a first reagent layer is formed; S6, coating the second reagent solution on the dried first reagent layer, and setting the wet film thickness, coating speed, and coating temperature of the coating. After the coating is completed, the second reagent layer is transferred to an oven at 50°C-70°C for drying for 10 minutes, and a second reagent layer with a dry film thickness of 50-200 μm is formed after drying. S7, cut the support matrix with the first reagent layer and the second reagent layer and the diffusion layer into 10*9.5mm 2 and assemble them according to the stacking sequence of the upper support layer, the diffusion layer, the first reagent layer, the second reagent layer, the support matrix and the lower support layer to form a total bilirubin dry detection analysis component.

10. The method for preparing a total bilirubin dry type detection and analysis component according to claim 9, characterized in that: Both ends of the upper supporting layer and the lower supporting layer are respectively provided with snap positions and buckles for connection and cooperation.

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