阅读说明：本技术 一种用于体外生化诊断的干化学法多层膜试剂 (Dry chemical method multi-layer membrane reagent for in vitro biochemical diagnosis ) 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体外生化诊断的干化学法多层膜试剂,包括上外框、样品滴加孔、过滤/扩散/反射多功能层、试剂层、透光层、透光孔、下外框,其中,所述上外框与下外框相互粘合,所述上外框的中心位置开有样品滴加孔,下外框中心位置开有透光孔,所述过滤/扩散/反射多功能层、试剂层、透光层从上到下依次设置于样品滴加孔和透光孔处的上外框和下外框之间,通过涂布工艺粘合在一起。通过本发明,以解决现有技术存在的液体试剂保存时间短、试剂稳定性差,以及以滤纸作为待测样品的扩散材料,扩散的均一性和反应的均一性较差,测试的准确性和精密度均较差的问题。(The invention provides a dry chemical method multi-layer membrane reagent for in vitro biochemical diagnosis, which comprises an upper outer frame, a sample dripping hole, a filtering/diffusing/reflecting multi-functional layer, a reagent layer, a light transmission hole and a lower outer frame, wherein the upper outer frame and the lower outer frame are bonded with each other, the sample dripping hole is formed in the center of the upper outer frame, the light transmission hole is formed in the center of the lower outer frame, and the filtering/diffusing/reflecting multi-functional layer, the reagent layer and the light transmission layer are sequentially arranged between the upper outer frame and the lower outer frame in the sample dripping hole and the light transmission hole from top to bottom and are bonded together through a coating process. The invention solves the problems of short preservation time of the liquid reagent, poor stability of the reagent, poor diffusion uniformity and reaction uniformity and poor test accuracy and precision by using the filter paper as the diffusion material of the sample to be tested in the prior art.)

1. The utility model provides a dry chemistry method multilayer membrane reagent for biochemical diagnosis in vitro, its characterized in that, includes frame (1), sample dropwise add hole (2), filters/diffusion/reflects many functional layers (3), reagent layer (4), euphotic layer (5), light trap (6), frame (7) down, wherein, go up frame (1) and frame (7) mutual bonding down, the central point of going up frame (1) puts and opens there is sample dropwise add hole (2), and frame (7) central point puts and opens there is light trap (6) down, filter/diffuse/reflect many functional layers (3), reagent layer (4), euphotic layer (5) from the top down set gradually between frame (1) and frame (7) down of sample dropwise add hole (2) and light trap (6) department, bond together through the coating technology.

2. The dry chemical multi-layer membrane reagent for in vitro biochemical diagnosis according to claim 1, wherein the lower outer frame (7) is attached with a glue layer for bonding the upper outer frame (1).

3. The dry chemical method multilayer membrane reagent for in vitro biochemical diagnosis according to claim 1, wherein the sample dropping hole (2) and the light transmission hole (6) are both circular holes with a diameter of 3-8 mm.

4. The dry chemical process multilayer membrane reagent for in vitro biochemical diagnosis according to claim 1, wherein the filtration/diffusion/reflection multifunctional layer (3) is an isotropic porous membrane integrating whole blood filtration, serum diffusion and light reflection functions, the porous membrane is composed of a hydrophilic polymer material and a white light reflecting material, the wet membrane thickness is 100 to 400 μm, the dry membrane thickness is 30 to 300 μm, and the pore size of the porous membrane ranges from 1 to 30 μm.

5. The dry chemical multi-layer membrane reagent for in vitro biochemical diagnosis according to claim 4, wherein the hydrophilic polymer material is cellulose acetate, and the white reflective material is one or a mixture of more reflective materials selected from titanium dioxide, barium sulfate, zinc oxide, lead oxide, diatomite, and microcrystalline cellulose.

6. The dry chemical multilayer membrane reagent for in vitro biochemical diagnosis according to claim 1, wherein the reagent layer (4) is composed of water-soluble materials and chemical reagents participating in the reaction, the wet membrane thickness is 50-400 μm, and the dry membrane thickness is 30-300 μm.

7. The dry chemical multi-layer membrane reagent for in vitro biochemical diagnosis according to claim 6, wherein the water-soluble substance of the reagent layer comprises one or more of gelatin, hydrophilic cellulose derivatives, dextran, agarose, polyvinyl alcohol, polyacrylamide, and the reagent participating in chemical reaction comprises one or more of enzymes, reaction substrates, buffers, cross-linking agents, surfactants, protective agents, and stabilizers.

8. The dry chemical method multilayer membrane reagent for in vitro biochemical diagnosis according to claim 1, wherein the light transmitting layer (5) is made of transparent plastic or glass having a light transmittance of 80% or more in the visible light range, and the thickness of the light transmitting layer is 30 to 300 μm.

Technical Field

The invention relates to the technical field of in-vitro clinical diagnosis reagents, in particular to a dry chemical method multi-layer membrane reagent for in-vitro biochemical diagnosis.

Background

In vitro diagnostic reagents are important auxiliary means in disease prevention, diagnosis and treatment. The biochemical diagnostic reagent is used for detecting biochemical indexes in vivo through various biochemical reactions or immunoreactions, and is mainly used for detecting by matching with instruments such as manual, semi-automatic and full-automatic biochemical analyzers and the like and measuring indexes such as enzymes, saccharides, lipids, proteins, non-protein nitrogen, inorganic elements, liver functions and the like. The existing biochemical analysis reagents are mainly liquid reagents of a wet chemical method. The liquid reagent has the disadvantages of short preservation time, poor reagent stability, easy environmental pollution caused by a large amount of waste liquid, and the like. The dry chemical method diagnostic reagent has the advantages of simple instrument operation, accurate result, low rechecking rate, no waste liquid, simple and convenient post-treatment, no pollution to water sources or soil, and suitability for occasions such as space stations, battlefields, fields or naval vessels, clinics or clinics, operating tables, home monitoring, patient beds and the like.

The dry chemical method in-vitro biochemical diagnostic reagent is mainly a product of foreign companies. The first type is a multilayer dry film represented by ausendor, in which a diffusion layer, a reflection layer, a reagent layer, etc. are coated on a transparent support layer by a film coating technique, and the change in optical density is measured from the reverse side of the support layer. The second type is dry chemical technology of kyoto corporation, japan, which consists of a plastic support layer, one end of which is a reaction zone comprising a sample layer and a reagent layer, the reagent layer using a fabric as a reagent carrier. Sample addition and testing were both performed in the upper part. The third kind is dry chemical test paper strip of Rogowski in Switzerland, which consists of magnetic tape cipher area, serum separating area and reaction area, and the whole blood is made to contact with reagent layer through transverse filtering with glass fiber. The multilayer film dry film of the Aosnto company has the advantages of accurate result, low rechecking rate, low cost and the like.

The domestic application of dry chemical in vitro biochemical diagnostic test, such as patent 200720072185.2, patent 200720073442.4 and patent 200910054558.7 applied by Shanghai Kehua bioengineering company, and patent 200610161430.7 applied by WushenRui biological products Limited company. The method used in these patents is to soak or spray the filter paper with the prepared reagent of the reaction solution, dry and then adhere to the plastic substrate for testing. The method is similar to pH test paper, filter paper is used as a diffusion material of a sample to be tested, the diffusion uniformity and the reaction uniformity are poor, the test accuracy and precision are poor, and the method is not suitable for clinical detection.

Disclosure of Invention

The invention provides a dry chemical method multi-layer membrane reagent for in vitro biochemical diagnosis, which aims to solve the problems of short preservation time of a liquid reagent and poor reagent stability in the prior art, and the problems of poor diffusion uniformity and reaction uniformity and poor test accuracy and precision by using filter paper as a diffusion material of a sample to be tested.

In order to solve the technical problems, the invention provides a dry chemical method multi-layer membrane reagent for in vitro biochemical diagnosis, which comprises an upper outer frame, a sample dripping hole, a filtering/diffusing/reflecting multi-functional layer, a reagent layer, a light transmitting hole and a lower outer frame, wherein the upper outer frame and the lower outer frame are mutually bonded, the sample dripping hole is formed in the central position of the upper outer frame, the light transmitting hole is formed in the central position of the lower outer frame, and the filtering/diffusing/reflecting multi-functional layer, the reagent layer and the light transmitting layer are sequentially arranged between the upper outer frame and the lower outer frame at the sample dripping hole and the light transmitting hole from top to bottom and are bonded together through a coating process.

The lower outer frame is attached with a glue layer for bonding the upper outer frame.

The sample dripping hole and the light transmission hole are round holes, and the diameter of each round hole is 3-8 mm.

The multifunctional filtering/diffusing/reflecting layer is an anisotropic porous membrane integrating whole blood filtering, serum diffusing and light reflecting functions, the porous membrane is composed of a hydrophilic polymer material and a white reflecting material, the thickness of the wet membrane is 100-400 mu m, the thickness of the dry membrane is 30-300 mu m, and the pore diameter range of the porous membrane is 1-30 mu m.

The hydrophilic polymer material is cellulose acetate, and the white reflective material is one or a mixture of more than one reflective material selected from titanium dioxide, barium sulfate, zinc oxide, lead oxide, diatomite and microcrystalline cellulose.

The reagent layer is composed of a water-soluble material and a chemical reagent participating in reaction, the wet film thickness is 50-400 mu m, and the dry film thickness is 30-300 mu m.

The water-soluble substance of the reagent layer comprises one or more of gelatin, hydrophilic cellulose derivative, dextran, agarose, polyvinyl alcohol and polyacrylamide, and the reagent participating in the chemical reaction comprises one or more of enzyme, reaction substrate, buffer solution, cross-linking agent, surfactant, protective agent and stabilizing agent.

The light transmitting layer is made of transparent plastic or glass with the light transmittance of more than 80% in the visible light range, and the thickness of the light transmitting layer is 30-300 mu m.

The invention has the following beneficial effects: the dry chemical method multi-layer membrane reagent for in vitro biochemical diagnosis can improve the accuracy and precision of detection, has traceability, is suitable for clinical detection, and can prolong the storage time of the reagent.

Drawings

FIG. 1 is a schematic structural view of a dry chemical method multi-layer membrane reagent for in vitro biochemical diagnosis according to the present invention.

Wherein, 1-upper outer frame, 2-sample dropping hole, 3-filtering/diffusing/reflecting multi-functional layer, 4-reagent layer, 5-euphotic layer, 6-euphotic hole and 7-lower outer frame.

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 fig. 1, the present invention provides a dry chemical method multi-layer membrane reagent for in vitro biochemical diagnosis, which comprises an upper outer frame 1, an upper outer frame sample dropping hole 2, a filtering/diffusing/reflecting multi-functional layer 3, a reagent layer 4, a light transmission layer 5, a lower outer frame light transmission hole 6 and a lower outer frame 7. Wherein, the upper frame 1 and the lower frame 7 are adhered to each other. The center of the upper outer frame 1 is provided with a round hole 2 for dripping samples, and the center of the lower outer frame 7 is provided with a light hole 6. The diffusion and reflection double functional layer 3, the reagent layer 4 and the light transmission layer 5 are bonded together through a coating process.

The main test samples of the dry chemical method multi-layer membrane reagent are whole blood, serum and urine.

Further, the sample may be applied to the surface of the filtration/diffusion/reflection multifunctional layer through the sample dropping hole of the upper outer frame. The filtering/diffusing/reflecting multifunctional layer functions to filter red blood cells, white blood cells, etc. in whole blood, uniformly diffuse and permeate serum to the next layer, and functions to reflect light. In some dry chemical multi-layer membrane reagents, depending on the test item, a reactive agent, such as an activator of reaction, a reagent for removing interfering substances, or an enzyme, may be added to the filtration/diffusion/reflection multi-functional layer. The diffusion/reflection multifunctional layer is made of hydrophilic high molecular materials such as cellulose acetate and white reflecting materials such as titanium dioxide, barium sulfate, zinc oxide, lead oxide, diatomite, microcrystalline cellulose or a mixture of the above reflecting materials. The filtering/diffusing/reflecting multi-functional layer is a layer of uniform and isotropic porous membrane, and the pore diameter range of the membrane is 1-30 mu m. The wet film thickness is 100-400 μm, the dry film thickness is 30-300 μm, and the effect is better within 50-200 μm.

Furthermore, the reagent layer is a region where a substance to be detected is subjected to a chemical reaction, the reagent layer can be a single layer or multiple layers, and according to the difference of the substance to be detected, a semi-permeable membrane can be added between the multiple reagent layers to selectively permeate part of products so as to react with the lower reagent layer. Each reagent layer is composed of water-soluble substances, such as gelatin, hydrophilic cellulose derivatives, dextran, agarose, polyvinyl alcohol, polyacrylamide and reagents participating in chemical reactions, such as enzymes, reaction substrates, buffers, cross-linking agents, surfactants, protective agents, stabilizing agents and the like. The wet film thickness is 50-400 μm, the dry film thickness is 30-300 μm, and the effect is better within 50-200 μm.

Further, the light-transmitting layer may be made of various glass or transparent plastic, such as polyethylene PE, polypropylene PP, polyester methylmethacrylate PMMA, polystyrene PS, polycarbonate PC, polyethylene terephthalate PET, etc., and PET is particularly preferable. The thickness of the transparent layer is 30 to 300 μm, preferably 100 to 200 μm. The light transmittance in the visible light range is more than 80%, and the optimal light transmittance is more than 95%. Both the incident light source and the reflected light can pass through the light-transmitting layer.

Further, the lower frame 7 is attached with a glue layer for adhering the upper frame 1. The upper and lower frames are for fixing the multi-layer film structure and preventing the reagent from being oxidized, so as to prolong the storage time of the reagent.

Furthermore, the sample dripping hole 2 and the light hole 6 are both round holes with the diameter of 3-8 mm.

Example 1: dry chemical method multi-layer membrane reagent for testing blood sugar

The light-transmitting layer 5 is a PET film having a thickness of 200 μm and has a light transmittance of 95% or more at 540 nm. The formulation of the reagent layer 4 is shown in table 1, and the prepared coating solution was uniformly coated on a PET film to a coating thickness of 150 μm. After drying, the filtering/diffusing/reflecting multifunctional layer 3 was coated thereon to a thickness of 300 μm, and the formulation of the filtering/diffusing/reflecting multifunctional layer 3 is shown in table 2. Drying, and cutting into 1cm²Then, white PET plastic sheets were used as the upper and lower frames 1 and 7.

Table 1 reagent layer formulation:

reagent	Concentration of
		Gelatin	300g/L
Glucose oxidase	77kU/L
		Peroxidase enzymes	360kU/L
1, 7-dihydroxynaphthalene	6.7g/L
		4-aminoantipyrines	11g/L

Table 2 filter/diffuser/reflector multifunctional layer formulation:

material	Concentration of
		Cellulose acetate	50g/L
Titanium dioxide	660g/L
		Surface active agent	10g/L

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.