阅读说明：本技术 一种用于总胆固醇定量检测的干片试剂 (Dry tablet reagent for quantitative detection of total cholesterol ) 是由 郑日记 黄利寒 甘斌 武伟民 李菁杨 于 2019-10-31 设计创作，主要内容包括：本发明提供一种用于总胆固醇定量检测的干片试剂,包括外壳,所述外壳内依次贴合有扩散层、试剂层、树脂层、基材,所述外壳上表面中心位置开有样品滴加孔,外壳底部中心位置开有透光孔,所述扩散层、试剂层、树脂通过涂布工艺从上到下依次分层贴合在基材上,所述扩散层主要由醋酸纤维素、硫酸钡、表面活性剂、氯化镁、甘油、聚乙二醇组成,所述试剂层主要由胆固醇氧化酶、胆固醇酯酶、过氧化物酶、磷酸缓冲液、聚丙烯酰胺、聚乙烯吡咯烷酮、明胶和酶保护剂EDTA组成。通过本发明,以解决现有技术存在的湿化学检测试剂不稳定、测试结果误差较大的问题。(The invention provides a dry sheet reagent for quantitatively detecting total cholesterol, which comprises a shell, wherein a diffusion layer, a reagent layer, a resin layer and a base material are sequentially attached in the shell, a sample dripping hole is formed in the central position of the upper surface of the shell, a light transmitting hole is formed in the central position of the bottom of the shell, the diffusion layer, the reagent layer and the resin are sequentially attached to the base material in a layering mode from top to bottom through a coating process, the diffusion layer mainly comprises cellulose acetate, barium sulfate, a surfactant, magnesium chloride, glycerol and polyethylene glycol, and the reagent layer mainly comprises cholesterol oxidase, cholesterol esterase, peroxidase, a phosphate buffer solution, polyacrylamide, polyvinylpyrrolidone, gelatin and an enzyme protective agent EDTA. The invention solves the problems of unstable wet chemical detection reagent and larger error of test result in the prior art.)

1. A dry sheet reagent for the quantitative determination of total cholesterol, which is characterized by comprising a shell (6), the diffusion layer (2), the reagent layer (3), the resin layer (4) and the base material (5) are sequentially adhered in the shell, the center of the upper surface of the shell (6) is provided with a sample dripping hole (7), the center of the bottom of the shell is provided with a light transmission hole (8), the diffusion layer (2), the reagent layer (3) and the resin (4) are laminated on the base material (5) from top to bottom in sequence through a coating process, the diffusion layer (2) mainly comprises cellulose acetate, barium sulfate, a surfactant, magnesium chloride, glycerol and polyethylene glycol, the reagent layer (3) mainly comprises cholesterol oxidase, cholesterol esterase, peroxidase, phosphate buffer, polyacrylamide, polyvinylpyrrolidone, gelatin and an enzyme protective agent EDTA.

2. The dry sheet reagent for the quantitative determination of total cholesterol according to claim 1, characterized in that the resin layer (4) is an aqueous epoxy resin.

3. The dry sheet reagent for the quantitative determination of total cholesterol according to claim 1, characterized in that the substrate (5) is pet material for providing a skeleton for supporting the reagent, and the transparency of the substrate is more than 80%.

4. The dry plate reagent for quantitatively detecting total cholesterol according to claim 1, wherein the housing (6) is made of plastic and is formed by laminating upper and lower layers of plastic and ultrasonically bonding the upper and lower layers of plastic, and is used for protecting the reagent from deformation caused by various external forces during transportation, storage and test, thereby avoiding damage to the reagent.

5. The dry plate reagent for quantitatively detecting total cholesterol according to claim 1, wherein the structure of the diffusion layer (2) is a three-dimensional network skeleton made of cellulose acetate, and the three-dimensional network skeleton is wrapped with the corresponding reagent.

6. The dry sheet reagent for quantitatively determining total cholesterol according to claim 1, wherein the reagent layer (3) is formed by coating a corresponding reagent with coagulated and dried gelatin as a carrier.

Technical Field

The invention relates to a total cholesterol detection reagent, in particular to a dry tablet reagent for quantitatively detecting total cholesterol.

Background

So-called "dry-patch reagents" are relatively conventional "wet chemistry". The method is a mode that liquid in a detected sample is used as a reaction medium, and an object to be detected directly reacts with a reagent fixed on a carrier, and the method is different from the traditional wet chemistry in the medium participating in the chemical reaction. With the development of techniques for separation, purification, storage, etc. of enzymes in biochemistry, the progress of sensor, photometer, and electrode techniques, and the popularization of computers, dry chemistry has also rapidly developed.

Dry chemistry has unique advantages over wet chemistry, and as for reagents, dry chemistry reagents do not need preparation before measurement, do not need calibration, and are stable, and whole blood can be directly measured; as for the instrument, a water supply and drainage system and a pure water system are not required, so that the operation is simple and the maintenance is easy.

Disclosure of Invention

The invention provides a dry sheet reagent for quantitatively detecting total cholesterol, which aims to solve the problems of unstable wet chemical detection reagent and larger error of a test result in the prior art.

In order to solve the technical problems, the invention provides a dry plate reagent for quantitatively detecting total cholesterol, which comprises a shell, wherein a diffusion layer, a reagent layer, a resin layer and a base material are sequentially attached to the shell, a sample dripping hole is formed in the center of the upper surface of the shell, a light transmission hole is formed in the center of the bottom of the shell, the diffusion layer, the reagent layer and the resin are sequentially attached to the base material in a layering mode from top to bottom through a coating process, the diffusion layer mainly comprises cellulose acetate, barium sulfate, a surfactant, magnesium chloride, glycerol and polyethylene glycol, and the reagent layer mainly comprises cholesterol oxidase, cholesterol esterase, peroxidase, a phosphate buffer solution, polyacrylamide, polyvinylpyrrolidone, gelatin and an enzyme protective agent EDTA.

The resin layer is a water-based epoxy resin.

The substrate is a pet material for providing a skeleton for supporting the reagent, the substrate having a transparency greater than 80%.

The shell is made of plastic and formed by ultrasonic bonding of upper and lower layers of plastic in a pressing mode, and is used for protecting the reagent from deformation caused by various external forces in the processes of transportation, storage and testing, so that the reagent is prevented from being damaged.

The structure of the diffusion layer takes cellulose acetate as a three-dimensional network framework, and the three-dimensional network framework is wrapped with a corresponding reagent.

The reagent layer is formed by wrapping corresponding reagents by taking coagulated and dried gelatin as a carrier.

The invention has the following beneficial effects: the dry tablet reagent is established on the basis of wet chemistry and used for a dry chemistry method for testing total cholesterol, has excellent performance, long storage time and accurate test result, can obviously improve various performance indexes of uric acid in the detection and storage processes, and makes up for the deficiency of wet chemistry.

Drawings

FIG. 1 is a schematic sectional view showing the structure of a dry plate reagent for quantitatively determining total cholesterol in accordance with the present invention.

FIG. 2 is a schematic diagram of the overall structure of the dry tablet reagent of the present invention.

Wherein, 1-sample, 2-diffusion layer, 3-reagent layer, 4-resin layer, 5-base material, 6-shell, 7-sample dropping hole and 8-light hole.

Detailed Description

In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention will be described in further detail below with reference to the accompanying drawings and specific embodiments.

As shown in figure 1, the invention provides a dry sheet reagent for quantitatively detecting total cholesterol, which comprises a shell 6, wherein a diffusion layer 2, a reagent layer 3, a resin layer 4 and a substrate 5 are sequentially attached in the shell, a sample dripping hole 7 is formed in the center of the upper surface of the shell 6, a light transmission hole 8 is formed in the center of the bottom of the shell, and the diffusion layer 2, the reagent layer 3 and the resin 4 are sequentially attached to the substrate 5 in a layering manner from top to bottom through a coating process.

Diffusion layer 2, diffusion layer are with cellulose acetate as three-dimensional network skeleton, wrap up corresponding reagents such as barium sulfate in the similar sponge of structure, and the main function includes even diffusion sample, filters macromolecular material and interfering substance in the sample, and diffusion layer mainly comprises cellulose acetate, barium sulfate, surfactant active, magnesium chloride, glycerine, polyethylene glycol etc.. The content of each diffusion layer component in each dry sheet is shown in table 1:

table 1: content of each component of diffusion layer

Name of reagent	Content per tablet (unit g)
		Cellulose acetate	0.002
Barium sulfate	0.02
		Surface active agent	0.00023
Magnesium chloride	0.00122
		Glycerol	0.0005
Polyethylene glycol	0.0005

The reagent layer 3 mainly comprises necessary biochemical reagents such as cholesterol oxidase, cholesterol esterase, peroxidase, phosphate buffer, polyacrylamide, polyvinylpyrrolidone, gelatin, enzyme protective agent EDTA and the like, and the reagent layer takes coagulated and dried gelatin as a carrier to wrap the corresponding reagent, and has a structure similar to jelly. The contents of the components of the reagent layer in each dry tablet are shown in table 2:

table 2: the content of each component of the reagent layer

Name of reagent	Each tablet content
		Cholesterol oxidase	0.6U
Cholesterol esterase	2.5U
		Peroxidase enzymes	3.7U
Phosphate buffer	50UL
		Polyacrylamide	0.03667G
Polyvinylpyrrolidone	0.0453G
		Gelatin	0.083G
EDTA	0.01667G
		Triton-100	0.00064G
Imidazole dyes	0.00025G

Resin layer 4, this resin layer can effectually make the reagent layer adhere to on the substrate, can reduce the requirement to the material and the performance of substrate, can use the substrate of not doing any processing, avoids handling the interference of substrate to detect reagent, and the resin layer is waterborne epoxy, and the structure is similar ordinary liquid glue, and the producer purchases and directly uses.

A substrate 5, which mainly provides a skeleton supporting the reagents, the transparency of which must be greater than 80%, the substrate being pet material, structured like a hard plastic paper sheet.

The shell 6, this shell layer is mainly that the protection reagent is in the transportation, stores, and the deformation that the in-process of test leads to because various power to avoid the damage of reagent, especially in last quick-witted test, no matter be automatic or semi-automatic send into the detection machine with the dry film reagent, the dry film inevitably receives corresponding power, the damage of the dry film of reagent can be avoided to the reagent shell, the shell is formed by upper and lower layer plastics pressfitting ultrasonic bonding, the overall structure of dry film reagent is as shown in figure 2.

The preparation method of the dry tablet reagent comprises the following steps:

1. a resin material is prepared to be coated on a base material.

2. The resin is coated with a reagent, and the reagent layer is coated by different coating methods according to needs and processes. The thickness and the roughness requirement on reagent layer are strict, and for guaranteeing the poor and stability between the batches of reagent, the thickness on reagent layer need strict control error, and wet thickness error control is in 200 +/-10 um, and dry thickness error need be controlled in 40 +/-3 um.

3. A diffusion layer is coated on the reagent. The coating thickness of the diffusion layer is 300 +/-20 um.

In summary, the reagent is coated on the transparent plastic film in the form of a film to be made into a dry sheet, and the dry sheet reagent corresponding to the concentration of the target substance is obtained by detecting the reflected light. The dry chemical reagent prepared by the method has excellent performance and long storage time.

The specific operation of the dry sheet reagent detection is as follows:

as shown in figure 1, 10ul of sample is added on the surface of the diffusion layer, the sample can be transversely and longitudinally uniformly diffused downwards through the diffusion layer and enters the reagent layer, total cholesterol of the target substance is catalyzed by cholesterol oxidase and cholesterol esterase to generate hydrogen peroxide, and the hydrogen peroxide reacts with imidazole dye under the catalysis of the peroxidase to generate a blue compound. After sample is added, light with the wavelength of 540nm is used for irradiating the position of the center of the base material, the change of the reflected light of the dried chip after sample addition is recorded, the density of the reflected light is in inverse proportion to the concentration of the total cholesterol of the target substance, and the corresponding concentration, namely the detection concentration of the total cholesterol corresponding to the sample is calculated by a computer. The equation for the reaction of the target substance total cholesterol with the corresponding substance in the dry tablet reagent is as follows:

in conclusion, the invention provides the dry tablet reagent which is established on the basis of wet chemistry and is used for the dry chemistry method for testing urea nitrogen, the dry tablet reagent has excellent performance, long storage time and accurate test result, can obviously improve each performance index of uric acid in the detection and storage processes, and makes up the deficiency of wet chemistry.

The above description is only an example of the present invention, and is not intended to limit the present invention, and it is obvious to those skilled in the art that various modifications and variations can be made in the present invention. Any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention should be included in the scope of the claims of the present invention.