Creatinine dry type detection analysis assembly and preparation method thereof

Abstract

The invention discloses a creatinine dry type detection analysis assembly, which comprises an upper support layer, a diffusion layer, a reagent layer, a support body and a lower support layer, the two ends of the upper support layer and the lower support layer are connected through buckles, the diffusion layer, the reagent layer and the support body are sequentially arranged between the upper support layer and the lower support layer, the middle position of the upper support layer is provided with a sample adding hole, and the sample adding hole is communicated with the sample adding hole. A test hole is formed in the middle of the lower supporting layer, the reagent layer comprises modified acrylic resin emulsion, modified gelatin, a buffer solution, creatininase, creatinase, sarcosine oxidase, peroxidase, 4-APP, an ALT indicator, a surfactant, soluble cellulose ether and an active material, and the invention further discloses a preparation method of the assembly. The creatinine dry type detection and analysis assembly does not need an organic solvent in the preparation process and is low in cost and environmentally friendly, and a dry type analysis reagent is good in stability, easy to store and keep, high in detection accuracy and wide in linear range.

Description

Technical Field

[0001] The present invention relates to the technical field of in vitro detection, and in particular to a creatinine dry type detection and analysis component and a preparation method thereof. Background Art

[0002] Dry chemical analysis 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 personnel to operate. At present, the commonly used dry analysis reagents continue the coating technology of photosensitive films, and the shielding layer, diffusion layer, reagent layer, color development layer, etc. are coated on the transparent support in sequence according to each functional layer, and the concentration of the object to be tested is detected by measuring the change of reflected light, which is easy to operate. However, since the dry analysis reagent needs to meet the requirements of shielding light sources, dispersing samples, and preventing oxidation and discoloration of the color development layer, its production process is complicated, and the production process requires coating of multiple layers of reagents, which is not only not environmentally friendly, but also causes the overall thickness of the reagent to be thick, and the reagent is easy to fall off and powder; and since the subsequent reflection photometry (Kubelka-Munk theory) needs to be used for testing, if the test area of ​​the test paper is small, it will cause the light receiving signal to be weak and the sensitivity is not enough. Therefore, the test area of ​​the existing dry analysis reagents is usually large, which further increases the cost. Summary of the invention

[0003] In order to solve the above problems, the present invention provides a creatinine dry detection analysis component and a preparation method thereof, so as to overcome the defects of the existing dry analysis reagent preparation process being complicated and costly.

[0004] The present invention adopts the following technical solutions:

[0005] A creatinine dry detection and analysis component comprises an upper support layer, a diffusion layer, a reagent layer, a support body and a lower support layer, wherein the upper support layer and the lower support layer are connected at both ends by snap fasteners, the diffusion layer, the reagent layer and the support body are sequentially arranged between the upper support layer and the lower support layer, a sample addition hole is arranged in the middle position of the upper support layer, a test hole is arranged in the middle position of the lower support layer, and the reagent layer comprises modified acrylic resin emulsion, modified gelatin, buffer, creatinase, creatinine enzyme, sarcosine oxidase, peroxidase, 4-APP, ALT indicator, surfactant, soluble cellulose ether and active material.

[0006] Furthermore, the modified acrylic resin emulsion is an emulsion-type thermosetting acrylic resin emulsion.

[0007] Furthermore, the buffer comprises any one of Tris buffer, glycine buffer, TES buffer, MES buffer solution, MOPS buffer or phosphate buffer, and its pH is 6-8.

[0008] Furthermore, the surfactant includes any one of sodium dodecylbenzene sulfonate, sodium hexadecyl sulfonate, tween-20, dodecylethoxysulfobetaine, tetradecylamidopropyl hydroxypropylsulfobetaine or tritonX-405, and the surfactant is added in an amount of 0.05 to 5 g per square meter of the reagent layer.

[0009] Furthermore, the soluble cellulose ether includes any one or more of methyl cellulose, hydroxyethyl methyl cellulose, carboxymethyl cellulose, ethyl cellulose, hydroxyethyl cellulose, hydroxypropyl methyl cellulose or phenyl cellulose, and the soluble cellulose ether is added in an amount of 1 to 20 g per square meter of the reagent layer.

[0010] Furthermore, the active material includes any one or more of diatomaceous earth, volcanic ash, silica gel, fly ash, microcrystalline cellulose, bentonite, titanium dioxide, and cellulose powder, and the active material is added in an amount of 1 to 40 g per unit square meter of the reagent layer.

[0011] Furthermore, the reagent layer also includes a defoamer, which includes any one or more of alcohols, fatty acids and fatty acid esters, amides, phosphates, silicones, polyethers, and polyether-modified polysiloxane defoamers, and the amount of the defoamer added per unit square meter of the reagent layer is 0.1 to 5 g.

[0012] Furthermore, the diffusion layer is a gauze with an opening rate of 500 or 600; the support is a PET material with a thickness of 100 to 300 um, including any one of polyethylene terephthalate, polybutylene terephthalate, and polyvinyl chloride.

[0013] Furthermore, the reagent layer is a transparent plastic substrate or a frosted plastic substrate with a transmittance of more than 80% under a wavelength of 300nm to 800nm, including any one of polyethylene terephthalate, polycarbonate, polypropylene, polyethylene, and polycarbonate sheets, and has a thickness of 30 to 200μm.

[0014] A method for preparing a creatinine dry type detection and analysis component, the method is used to prepare the creatinine dry type detection and analysis component as described above, the method comprising the following steps:

[0015] S1, preparing a TES buffer solution with a pH of 6 to 8, adding modified acrylic resin emulsion, modified gelatin, surfactant, soluble cellulose ether and active material to the TES buffer solution in sequence, stirring evenly, controlling the stirring speed at 600 r / min to 1000 r / min, and stirring time for 2 h;

[0016] S2, continue to add creatinase, creatinase, sarcosine oxidase, peroxidase, 4-APP and ALT indicator to step S1, and after the addition is completed, the stirring speed is adjusted to 400r / min-500r / min, and the stirring time is 2h;

[0017] S3, moving the reagent layer slurry stirred evenly in step S2 onto the support for coating, the coating temperature is 27.5°C, the coating wet film thickness is 100-500 μm, and drying is carried out at a temperature range of 50°C-70°C, and the dry film thickness after drying is 50-200 μm;

[0018] S4. Cut the reagent sheet and diffusion layer gauze dried in step S3 into small pieces respectively, use black PET plastic card sleeves as upper support layer and lower support layer, fix and assemble them into a dry detection and analysis component.

[0019] After adopting the above technical solution, the present invention has the following advantages compared with the background technology:

[0020] 1. The present invention adopts a combination of synthetic acrylic resin emulsion, modified gelatin and polymer materials to solve the problem of low-temperature storage. The acrylic resin emulsion can improve the background and shield part of the light source without affecting the color development and reflectivity test, so that the overall linear range of the test is widened.

[0021] 2. The present invention forms a reagent layer by connecting soluble cellulose ether and active materials under the joint action of surfactant and defoamer. The reagent layer is coated with only one layer, which solves the problem of multi-layer coating, easy shedding and powder loss in the prior art, simplifies the preparation process and reduces the cost;

[0022] 3. The present invention uses a gauze mesh as a diffusion layer, which is environmentally friendly and harmless, and can achieve rapid penetration and accurate detection of samples containing creatinine.

[0023] 4. The creatinine dry detection and analysis component of the present invention does not require organic solvents during the preparation process and is environmentally friendly. The dry analysis reagent has good stability, is easy to store and preserve, has high detection accuracy, and a wide linear range. BRIEF DESCRIPTION OF THE DRAWINGS

[0024] Figure 1 It is a structural schematic diagram of the creatinine dry detection and analysis component of the present invention;

[0025] Figure 2 This is a linear correlation diagram of the test results of Example 2;

[0026] Figure 3 This is a linear correlation diagram of the test results of Example 3;

[0027] Figure 4 This is a linear correlation diagram of the test results of Example 4.

[0028] Description of reference numerals:

[0029] 1. Upper support layer; 2. Diffusion layer; 3. Reagent layer; 4. Support body; 5. Lower support layer; 6. First card position; 7. Second card position; a. Sample loading hole; b. Test hole. DETAILED DESCRIPTION

[0030] 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 intended to limit the present invention.

[0031] Embodiment 1

[0032] like Figure 1 As shown, a creatinine dry detection and analysis component includes an upper support layer 1, a diffusion layer 2, a reagent layer 3, a support body 4 and a lower support layer 5, wherein the upper support layer 1 and the lower support layer 5 are respectively connected at both ends by a first clamping position 6 and a second clamping position buckle, the diffusion layer 2, the reagent layer 3 and the support body 4 are sequentially arranged between the upper support layer 1 and the lower support layer 5, a sample addition hole a is provided in the middle position of the upper support layer 1, a test hole b is provided in the middle position of the lower support layer 5, and the reagent layer 3 includes an acrylic resin emulsion modified, modified gelatin, a buffer, creatinase, creatinine enzyme, sarcosine oxidase, peroxidase, 4-APP, an ALT indicator, a surfactant, a soluble cellulose ether, an active material and a defoaming agent.

[0033] The modified acrylic resin emulsion is an aqueous acrylic resin emulsion, which includes two types: thermosetting acrylic ester emulsion and thermoplastic acrylic ester emulsion. Thermosetting acrylic ester emulsion is divided into emulsion type, water dilutable and aqueous solution. The modified acrylic resin emulsion of this embodiment adopts an emulsion type thermosetting acrylic resin emulsion, and the added amount per unit square meter on the reagent layer 3 is 1 to 30g, preferably 8g per unit square meter. It is characterized by good film-forming property, strong adhesion, good shielding property, fast drying speed, green and environmentally friendly, and has advantages in reagent preparation.

[0034] The preparation method of the above modified gelatin is as follows:

[0035] (1) dissolving 3,4-dihydroxybenzoic acid in ethyl acetate, adding acetyl chloride to carry out heating reflux reaction, and removing ethyl acetate to obtain a first modifier;

[0036] (2) uniformly dispersing the first modifier in trichloropropane, then adding dimethyl sulfoxide to carry out reflux reaction, then distilling and separating the dimethyl sulfoxide, and obtaining the second modifier after drying and grinding;

[0037] (3) Dissolve gelatin in distilled water, adjust the pH to 8.5 with NaOH solution, add the second modifier, stir well, precipitate and wash with sulfuric acid solution, melt with distilled water, adjust the pH to 7.0 with NaOH solution, and freeze-dry to obtain the product.

[0038] The modified gelatin has a good gelling effect and can be stored at room temperature. The amount added to the reagent layer 3 is 1 to 15 g per square meter, preferably 6 g per square meter.

[0039] The buffer used is Tris buffer, and its pH is 7.

[0040] The surfactant is dodecylethoxysulfobetaine, and the amount of the surfactant added per unit square meter of the reagent layer 3 is 2.5 g.

[0041] The soluble cellulose ether is hydroxyethyl methyl cellulose, and the amount of the soluble cellulose ether added to the reagent layer 3 per unit square meter is 10 g.

[0042] The active material is silica gel and fly ash, and the amount of the active material added to the reagent layer 3 per unit square meter is 20 g.

[0043] The defoaming agent is alcohol, and the amount of the defoaming agent added per unit square meter of the reagent layer 3 is 2.5 g.

[0044] In this embodiment, the diffusion layer 2 is a gauze with an opening rate of 500; the support body 4 is a PET material with a thickness of 200um, that is, polyethylene terephthalate; the reagent layer 3 is polyethylene terephthalate with a transmittance of 85% under a wavelength of 500nm, and its thickness is 120um.

[0045] This embodiment takes the above-mentioned creatinine dry type detection and analysis component as an example to further illustrate its preparation method, which comprises the following steps:

[0046] S1, prepare TES buffer solution, weigh TES powder, BSA, sucrose, activator and other reagents, add them to the aqueous solution, wait for complete dissolution, adjust the pH to 7 with a pH meter, add modified acrylic resin emulsion, modified gelatin, surfactant, soluble cellulose ether and active material to the TES buffer solution in sequence, stir evenly with a spiral stirrer, control the stirring speed at 800r / min, and stir for 2h;

[0047] When adding modified acrylic resin emulsion, it is necessary to stir while adding to prevent the emulsion from reacting with other substances in the slurry and precipitating particles. When adding soluble cellulose ether and active materials, it is also necessary to add small amounts and 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 a MYP2011-150 electric agitator.

[0048] S2, continue to add creatinase, creatinase, sarcosine oxidase, peroxidase, 4-APP and ALT indicator to step S1, and after adding, the stirring speed is adjusted to 450r / min to prevent the enzyme from being destroyed and the stirring time is 2h;

[0049] S3, the reagent layer 3 slurry stirred evenly in step S2 is moved to the support 4 for coating, the coating machine used is a heating scraper coater of model PW-TB100, the coating temperature is 27.5°C, the coating wet film thickness is 300um, and it is dried at a temperature range of 60°C, and the dry film thickness after drying is 155-165um;

[0050] During coating, place the frosted 125um thick support 4 on the coating machine, press both ends tightly, return the micrometer to zero, set the wet film thickness of the coating machine to 315um, pour the reagent layer 3 slurry to the bottom of the scraper mouth, press the coating button, the scraper starts coating, and the coating speed is controlled at 30cm / min.

[0051] S4, cut the reagent sheet and the diffusion layer 2 gauze dried in step S3 into 10*9.5mm 2 The small pieces are fixed and assembled into a dry detection and analysis component using a black PET plastic card sleeve as an upper support layer 1 and a lower support layer 5.

[0052] The creatinine dry detection and analysis component prepared above can be used to determine creatinine in serum, and the serum can be a Landau quality control serum sample or a serum sample filtered from whole blood. When in use, the serum to be tested is dripped onto the diffusion layer 2 through the sample addition hole a, and the serum to be tested quickly diffuses to the reagent layer 3 through the diffusion layer 2, and a biochemical reaction occurs in the reagent layer 3. The specific detection method is as follows:

[0053] Vertically add 10ul of the serum sample to be tested to the sample well a, and the sample diffuses from the diffusion layer 2 to the reagent layer 3, and incubates at about 37°C for 5 minutes. In this process, the endogenous creatine in the sample is oxidized, and the creatinine is hydrolyzed into creatine under the action of creatinase, and the creatine is converted into sarcosine and urea by creatinase. Sarcosine is oxidized into glycine, formaldehyde and hydrogen peroxide under the action of sarcosine oxidase. Hydrogen peroxide is oxidized into a blue quinone compound with the color indicator under the catalysis of peroxidase. At a wavelength of 640nm, the change in reflectivity is measured by reflectance spectrophotometry, and the concentration of creatinine is calculated by the change in reflectivity. The equation for the reaction of the corresponding substances in the creatinine dry analysis reagent is as follows:

[0054]

[0055] The present invention optimizes and innovates the formula, coating process and reagent cost, uses synthetic acrylic resin emulsion, combines synthetic acrylic resin emulsion with modified gelatin and polymer materials, connects soluble cellulose ether and active adsorption materials, and prepares a diffusion block and a dry analytical reagent with uniform penetration, which realizes room temperature storage, simplifies the coating process, solves the problem of easy powdering and falling off of the reagent, reduces the production cost, and also solves the problem of organic reagent pollution and harm to the human body in the diffusion layer 2. The dry analytical reagent for creatinine detection has high test result accuracy and wide measurement range, meets the needs of instant diagnosis and on-site diagnosis, and has great clinical application value.

[0056] Embodiment 2

[0057] Prepare the buffer solution and adjust the pH to 7.0. Add the components of the reagent group to the buffer solution in turn. Stir with a spiral stirrer and control the speed at 700r / min. When the solution is stirred evenly, place the frosted 125um thick PC film support 4 on the coating machine, press both ends, return the micrometer to zero, set the wet film thickness of the coating machine to 315um, pour the reagent layer slurry to the bottom of the scraper mouth, press the coating button, and the scraper will start coating. The coating speed is controlled at 30cm / min. Place the coated reagent in a 50℃ oven for constant temperature drying. The thickness after drying reaches 155~165um. Cut the dried reagent layer with a long strip cutter, and the cutting specifications are 294*10mm. Then cut the gauze into the same specifications and sizes, and then cut them into 10*9.5mm respectively. 2 Finally, a black PET card sleeve is used as the upper support layer 1 and the lower support layer 5 to fix the reagent layer and assemble it into a dry analytical reagent, which is then used for determination.

[0058] Table 1 Reagent layer formula

[0059]

[0060]

[0061] 25 different quality control serum samples were prepared in proportion with Landau serum, and the correlation of multiple quality control samples was analyzed using the method of the present invention and the biochemical analyzer test method. The results are as follows Figure 2 As shown, the test results of the method of the present invention and the test results of the biochemical analyzer method show a good linear correlation.

[0062] Embodiment 3

[0063] Prepare the buffer solution and adjust the pH to 7.2. Add the components of the reagent group to the buffer solution in turn. Stir with a spiral stirrer at a speed of 500r / min. When the solution is stirred evenly, place the colorless, transparent, 125um thick PET film support 4 on the coating machine, press both ends, return the micrometer to zero, set the coating machine wet film thickness to 340um, pour the reagent layer slurry to the bottom of the scraper, press the coating button, and the scraper starts coating. The coating speed is controlled at 50cm / min. Place the coated reagent in a 60℃ oven for constant temperature drying. The thickness after drying reaches 152-162μm. Cut the dried reagent layer with a long strip cutter, and the cutting specification is 294*10mm. The gauze was then cut into the same size, and then cut into small pieces of 10*9.5 mm2. Finally, the reagent layer was fixed and assembled into a dry analytical reagent using a black PET card sleeve as the upper support layer 1 and the lower support layer 5, and then used for determination.

[0064] Table 2 Reagent layer formula

[0065] Reagents <![CDATA[Coating amount / m 2 > MES (Tris(hydroxymethyl)ethylaminoethanesulfonic acid) PH7.2 Modified acrylic resin emulsion 15g Modified Gelatin 5g Methylcellulose 15g Myristylamidopropyl Hydroxypropyl Sulfobetaine 50mg Amide defoamer 2ml BSA (Bovine Serum Albumin) 50mg Cellulose powder 15g Volcanic ash 5g Creatinase 0.2u Creatinase 200u Sarcosine oxidase 7u POD enzyme (peroxidase) 0.7u 4-AAP (4-aminoantipyrine) 50mg Color indicator 200mg

[0066] 25 different quality control serum samples were prepared in proportion with Landau serum, and the correlation of multiple test samples was analyzed using the method of the present invention and the biochemical analyzer test method. Figure 3 As shown, the test results of the method of the present invention and the test results of the biochemical analyzer method show a good linear correlation.

[0067] Embodiment 4

[0068] Prepare the buffer solution and adjust the pH to 7.5. Add the components of the reagent group to the buffer solution in turn. Stir with a spiral stirrer and control the speed at 600r / min. When the solution is stirred evenly, place the frosted 125um thick PC film support 4 on the coating machine, press both ends, return the micrometer to zero, set the coating machine wet film thickness to 360um, pour the reagent layer slurry to the bottom of the scraper mouth, press the coating button, and the scraper starts coating. The coating speed is controlled at 80cm / min. Place the coated reagent in a 70℃ oven for constant temperature drying. The thickness after drying reaches 150-160μm. Cut the dried reagent layer with a long strip cutter, and the cutting specification is 294*10mm. The gauze was then cut into the same size, and then cut into small pieces of 10*9.5 mm2. Finally, the reagent layer was fixed and assembled into a dry analytical reagent using a black PET card sleeve as the upper support layer 1 and the lower support layer 5, and then used for determination.

[0069] Table 3 Reagent layer formula

[0070] Reagents <![CDATA[Coating amount / m 2 > MES (Tris(hydroxymethyl)ethylaminoethanesulfonic acid) PH7.5 Modified acrylic resin emulsion 9g Modified Gelatin 3g Carboxymethyl cellulose 12g Sodium dodecylbenzenesulfonate 100mg Silicone defoamer 1ml BSA (Bovine Serum Albumin) 100mg Microcrystalline Cellulose 12g Bentonite 5g Creatinase 0.2u Creatinase 200u Sarcosine oxidase 7u POD enzyme (peroxidase) 0.7u 4-AAP (4-aminoantipyrine) 50mg Color indicator 200mg

[0071] 25 different quality control serum samples were prepared in proportion with Landau serum, and the correlation of multiple test samples was analyzed using the method of the present invention and the biochemical analyzer test method. Figure 4 As shown, the test results of the method of the present invention and the test results of the biochemical analyzer method show a good linear correlation.

[0072] 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 creatinine dry detection and analysis component, characterized in that: The invention comprises an upper support layer, a diffusion layer, a reagent layer, a support body and a lower support layer, wherein the upper support layer and the lower support layer are connected at two ends by snap fasteners, the diffusion layer, the reagent layer and the support body are sequentially arranged between the upper support layer and the lower support layer, a sample addition hole is arranged in the middle of the upper support layer, a test hole is arranged in the middle of the lower support layer, and the reagent layer comprises modified acrylic resin emulsion, modified gelatin, buffer, creatinase, creatinase, sarcosine oxidase, peroxidase, 4-APP, ALT indicator, surfactant, soluble cellulose ether and active material.

2. A creatinine dry type detection and analysis component as claimed in claim 1, characterized in that: The modified acrylic resin emulsion is an emulsion-type thermosetting acrylic resin emulsion.

3. A creatinine dry type detection and analysis component as claimed in claim 2, characterized in that: The buffer comprises any one of Tris buffer, glycine buffer, TES buffer, MES buffer solution, MOPS buffer or phosphate buffer, and its pH is 6-8.

4. A creatinine dry type detection and analysis component as claimed in claim 3, characterized in that: The surfactant includes any one of sodium dodecylbenzene sulfonate, sodium hexadecyl sulfonate, tween-20, dodecylethoxysulfobetaine, tetradecylamidopropyl hydroxypropylsulfobetaine or tritonX-405, and the surfactant is added in an amount of 0.05 to 5 g per square meter of the reagent layer.

5. A creatinine dry type detection and analysis component as claimed in claim 4, characterized in that: The soluble cellulose ether includes any one or more of methyl cellulose, hydroxyethyl methyl cellulose, carboxymethyl cellulose, ethyl cellulose, hydroxyethyl cellulose, hydroxypropyl methyl cellulose or phenyl cellulose, and the soluble cellulose ether is added in an amount of 1 to 20 g per square meter of the reagent layer.

6. A creatinine dry type detection and analysis component as claimed in claim 5, characterized in that: The active material comprises any one or more of diatomaceous earth, volcanic ash, silica gel, fly ash, microcrystalline cellulose, bentonite, titanium dioxide, and cellulose powder, and the amount of the active material added per unit square meter of the reagent layer is 1 to 40 g.

7. A creatinine dry type detection and analysis component as claimed in claim 6, characterized in that: The reagent layer also includes a defoamer, which includes any one or more of alcohols, fatty acids and fatty acid esters, amides, phosphates, silicones, polyethers, and polyether-modified polysiloxane defoamers, and the amount of the defoamer added per unit square meter of the reagent layer is 0.1 to 5 g.

8. A creatinine dry type detection and analysis component as claimed in claim 7, characterized in that: The diffusion layer is a gauze with an opening rate of 500 or 600; the support is a PET material with a thickness of 100 to 300 um, including any one of polyethylene terephthalate, polybutylene terephthalate, and polyvinyl chloride.

9. A creatinine dry type detection and analysis component as claimed in claim 8, characterized in that: The reagent layer is a transparent plastic substrate or a frosted plastic substrate with a light transmittance of more than 80% under a wavelength of 300nm to 800nm, including any one of polyethylene terephthalate, polycarbonate, polypropylene, polyethylene and polycarbonate sheets, and has a thickness of 30 to 200μm.

10. A method for preparing a creatinine dry detection and analysis component, characterized in that: The method is used to prepare the creatinine dry detection and analysis component as described in any one of claims 1 to 9, and the method comprises the following steps: S1, preparing a TES buffer solution with a pH of 6 to 8, adding modified acrylic resin emulsion, modified gelatin, surfactant, soluble cellulose ether and active material to the TES buffer solution in sequence, stirring evenly, controlling the stirring speed at 600 r / min to 1000 r / min, and stirring time for 2 h; S2, continue to add creatinase, creatinase, sarcosine oxidase, peroxidase, 4-APP and ALT indicator to step S1, and after the addition is completed, the stirring speed is adjusted to 400r / min-500r / min, and the stirring time is 2h; S3, moving the reagent layer slurry stirred evenly in step S2 onto the support for coating, the coating temperature is 27.5°C, the coating wet film thickness is 100-500 μm, and drying is carried out at a temperature range of 50°C-70°C, and the dry film thickness after drying is 50-200 μm; S4. Cut the reagent sheet and diffusion layer gauze dried in step S3 into small pieces respectively, use black PET plastic card sleeves as upper support layer and lower support layer, fix and assemble them into a dry detection and analysis component.

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