CN110794147A - Albumin quantitative determination dry tablet for in vitro diagnosis - Google Patents
Albumin quantitative determination dry tablet for in vitro diagnosis Download PDFInfo
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- CN110794147A CN110794147A CN201911056563.1A CN201911056563A CN110794147A CN 110794147 A CN110794147 A CN 110794147A CN 201911056563 A CN201911056563 A CN 201911056563A CN 110794147 A CN110794147 A CN 110794147A
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
- G01N33/6803—General methods of protein analysis not limited to specific proteins or families of proteins
- G01N33/6827—Total protein determination, e.g. albumin in urine
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/52—Use of compounds or compositions for colorimetric, spectrophotometric or fluorometric investigation, e.g. use of reagent paper and including single- and multilayer analytical elements
- G01N33/525—Multi-layer analytical elements
- G01N33/526—Multi-layer analytical elements the element being adapted for a specific analyte
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
- G01N2333/435—Assays involving biological materials from specific organisms or of a specific nature from animals; from humans
- G01N2333/76—Assays involving albumins other than in routine use for blocking surfaces or for anchoring haptens during immunisation
- G01N2333/765—Serum albumin, e.g. HSA
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Abstract
The invention provides an albumin quantitative detection dry sheet for in vitro diagnosis, which comprises an upper shell with a sample dripping hole, a diffusion layer, a reagent layer, a resin layer, a support layer and a lower shell with a light hole, wherein the upper shell and the lower shell are mutually attached, the sample dripping hole is formed in the center of the upper shell, the light hole is formed in the center of the lower shell, the diffusion layer, the reagent layer, the resin layer and the support layer are sequentially arranged between the upper shell and the lower shell from top to bottom and are sequentially laminated and attached together from top to bottom through a coating process, and the reagent layer comprises a pigment, a buffer solution and a hydrophilic polymer. The invention solves the problems that the prior fluorescence photometry technology has higher requirements on instruments, a spectrophotometer which can be used for detection is expensive in a liquid phase, and the result can not be stored for a long time.
Description
Technical Field
The invention relates to an in vitro diagnosis detection reagent, in particular to an albumin quantitative detection dry tablet for in vitro diagnosis.
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.
Compared with wet chemistry, the dry chemistry analysis method has the advantages of corresponding detection instruments, simple operation, short detection time, accurate obtained result and the like. The application range of a dry chemical test paper method in clinical examination is obviously expanded, more and more biochemical examination items can be carried out, VITORS350 proposed by Olson corporation in America is suitable for a full-automatic analysis system for emergency treatment and conventional biochemistry, samples such as serum, plasma, whole blood, urine, cerebrospinal fluid and the like can be detected, the detection items comprise combined items of liver function, kidney function, myocardial enzyme, blood fat, protein, ions and the like or any single item, the rapid, accurate and accurate result can be really realized, the clinical accurate, rapid and flexible diagnosis requirements can be fully met, but a matched instrument is overlarge, and the detection can not be carried out in real time.
With the progress of biotechnology, the immune dry sheet containing enzyme markers and fluorescence markers, especially colloidal gold or selenium-labeled antibodies (or antigens) developed by utilizing technologies such as immunoosmosis, immunochromatography and the like can be used for analytical determination of troponin, special proteins, hormones, certain therapeutic drugs, virus antibodies or antigens and the like, the multilayer dry sheet adopting a fluorescence photometry and a matched instrument thereof are also available and are used more and more, and a vaginitis quintuplet detection reagent of Henan atlas biology company is used for detecting specific biochemical markers by a dry chemical enzyme method and is used for diagnosis of gynecological diseases. After more than 20 years of development, dry chemistry analysis techniques have been widely used to examine various aspects of medicine, including routine biochemistry, endocrine hormone, toxin drug concentration analysis, and special protein immunoassays. However, the requirement of the fluorescence photometry technology on instruments is high, a spectrophotometer capable of being used for detection is expensive in a liquid phase, and the result cannot be stored for a long time; the colloidal gold technology is used as a primary screening diagnostic reagent for the auxiliary treatment of disease diagnosis because the quality of a product is greatly different, the quality cannot be controlled, the quality cannot be ensured, and the phenomenon of the back zone exists, and a specimen needs to be diluted and tested, can only be qualitative and cannot be quantitative.
Disclosure of Invention
The invention provides an albumin quantitative detection dry sheet for in vitro diagnosis, which aims to solve the problems that the requirement of a fluorescence photometry technology on an instrument in the prior art is higher, a spectrophotometer capable of being used for detection is expensive in a liquid phase, and the result cannot be stored for a long time; and the colloidal gold technology has the problems that the product quality is relatively large in difference, the quality cannot be controlled, the quality cannot be ensured, the phenomenon of back zone exists, a sample needs to be diluted and tested, and only qualitative and quantitative measurement can be carried out.
In order to solve the technical problem, the invention provides an albumin quantitative detection dry sheet for in vitro diagnosis, which comprises an upper shell with a sample dripping hole, a diffusion layer, a reagent layer, a resin layer, a support layer and a lower shell with a light hole, wherein the upper shell and the lower shell are mutually attached, the sample dripping hole is formed in the center of the upper shell, the light hole is formed in the center of the lower shell, the diffusion layer, the reagent layer, the resin layer and the support layer are sequentially arranged between the upper shell and the lower shell from top to bottom and are sequentially laminated and attached together from top to bottom through a coating process, and the reagent layer comprises a pigment, a buffer solution and a hydrophilic polymer.
The pigment is an acid-base indicator capable of combining with albumin to generate color change, and the acid-base indicator comprises one of bromocresol green, bromocresol purple, bromothymol blue, phenol red, cresol red, cresolphthalein, indigo carmine, methyl red and methyl orange; the hydrophilic polymer comprises one or more of gel, gelatin derivative, colla Corii Asini, polyvinyl alcohol, polyvinylpyrrolidone, and polyacrylamide; the buffer solution is acetic acid buffer solution or citric acid buffer solution.
The wet film thickness of the reagent layer is 100-300 mu m, and the dry film thickness is 40-200 mu m.
The diffusion layer mainly comprises a hydrophilic high polymer material and a white reflective material, wherein the hydrophilic high polymer material is cellulose acetate or carboxymethyl cellulose, and the white reflective material is one or a mixture of more than one of titanium dioxide, barium sulfate, polystyrene and microcrystalline cellulose; the wet film thickness of the diffusion layer is 300-500 μm, and the dry film thickness is 70-300 μm.
The material of the supporting layer can be composed of various transparent plastics; the thickness of the support layer is 30-200 mu m; the light transmittance of the support layer in the visible light range is more than 85%, and both incident light source and reflected light can pass through the support layer.
The transparent plastic is made of Polyethylene (PE), polypropylene (PP), Polyester Methyl Methacrylate (PMMA), Polystyrene (PS), Polycarbonate (PC) or polyethylene terephthalate (PET).
The resin layer is arranged between the supporting layer and the reagent layer coating film, the supporting layer and the reagent layer are adhered together by the resin layer with adhesion, and the resin layer is a novel resin system taking water instead of an organic solvent as a dispersion medium.
The material of the resin layer is one of epoxy resin, alkyd resin, polyester resin, polyurethane resin and modified polybutadiene resin; the wet film thickness of the resin layer is 10-100 μm, and the dry film thickness is 2-50 μm.
The upper shell and the lower shell are mainly made of plastic materials, and the upper shell layer and the lower shell layer are attached together in an ultrasonic attaching mode.
The invention has the following beneficial effects: the albumin quantitative detection dry tablet for in vitro diagnosis provided by the invention is a dry chemical method multi-layer membrane reagent dry tablet for in vitro diagnosis, has the advantages of rapid diagnosis, simple operation, cost saving and the like, can improve the accuracy, precision and sensitivity of detection, and has traceability, stable reagent and long storage time.
Drawings
FIG. 1 is a schematic structural diagram of a quantitative albumin detection dry plate for in vitro diagnosis according to an embodiment of the present invention.
Wherein, 1-upper shell, 2-diffusion layer, 3-reagent layer, 4-resin layer, 5-support layer, 6-lower 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 an albumin quantitative determination dry plate for in vitro diagnosis, which comprises an upper shell 1 with a sample dripping hole, a diffusion layer 2, a reagent layer 3, a resin layer 4, a support layer 5 and a lower shell 6 with a light hole, wherein the upper shell 1 and the lower shell 6 are mutually attached, the central position of the upper shell is provided with the sample dripping hole 7, the central position of the lower shell is provided with the light hole 8, and the diffusion layer 2, the reagent layer 3, the resin layer 4 and the support layer 5 are sequentially arranged between the upper shell 1 and the lower shell 6 from top to bottom and are sequentially attached together in a layered manner from top to bottom through a coating process.
The invention consists of a module containing dry chemical reagents attached to a rigid plastic strip. The color of the module changes due to the reaction between various chemical examination contents in the blood and dry chemical reagents, and the color depth or the color change speed of the module is in proportional relation with the concentration of corresponding chemical components in the blood and urine.
Further, the albumin multilayer film dry sheet can be used for quantitatively measuring the concentration of albumin in serum and plasma. The albumin dry sheet is a multilayer analysis component coated on a polyester substrate, a sample is dripped to the surface of a diffusion layer through a sample dripping hole of an upper shell, is uniformly distributed to a reagent layer below through the diffusion layer, liquid permeates into the reagent layer, and a dye permeates into the diffusion layer and is combined with albumin in the sample, and the combination with the albumin can shift the wavelength of the maximum reflected light of the free dye. The resulting colored complex is measured by reflectance photometry. The amount of albumin bound dye is directly proportional to the concentration of albumin in the sample. The specific reaction steps are as follows:
albumin + bromocresol Green → BCG-Albumin Complex
The diffusion layer mainly comprises hydrophilic high polymer materials and white reflecting materials, the hydrophilic high polymer materials comprise cellulose acetate, carboxymethyl cellulose and the like, the white reflecting materials comprise titanium dioxide, barium sulfate, polystyrene, microcrystalline cellulose or mixtures of the above reflecting materials, and the reflecting materials mainly provide a background for reflection measurement; the diffusion layer has the main functions of uniformly diffusing a sample, filtering macromolecular substances and interfering substances in the sample and protecting a reagent layer; the wet film thickness of the diffusion layer is 300-500 μm, and the dry film thickness is 70-300 μm, and the effect is better within 100-250 μm. The reagent layer is a region where a substance to be detected is subjected to chemical reaction, the reagent layer comprises necessary biological agents such as necessary pigment, buffer solution and hydrophilic polymer, the pigment is an acid-base indicator capable of combining with albumin to generate color change, such acid-base indicator can be indexes such as indigoid dye (such as bromocresol green, bromocresol purple, bromothymol blue, phenol red, cresol red, cresolphthalein, indigo carmine and the like) or azo dye (methyl red, methyl orange and the like), and the indexes are most ideal with the bromocresol green and the bromocresol purple. Hydrophilic polymers include gelatin (e.g., acid-washed gelatin, deionized gelatin), gelatin derivatives (e.g., titanates, methyl hydroxymethyl acrylate, etc.), gelatin, polyvinyl alcohol, polyvinylpyrrolidone, polyacrylamide, and the like. The buffer solution is pH buffer solution, the pH buffer solution is added to keep the indicator in a corresponding color development pH value range, and acetic acid buffer solution or citric acid buffer solution can be selected. The wet film thickness of the reagent layer is 100-300 μm, and the dry film thickness is 40-200 μm, and the effect is better within 50-150 μm. The supporting layer can be made of various transparent plastics such as Polyethylene (PE), polypropylene (PP), Polyester Methyl Methacrylate (PMMA), Polystyrene (PS), Polycarbonate (PC), polyethylene terephthalate (PET) and the like, and preferably PET; the thickness of the supporting layer is 30-200 μm, preferably 50-150 μm; the light transmittance of the visible light range is more than 85 percent, the optimal light transmittance is more than 95 percent, and the incident light source and the reflected light can both pass through the supporting layer. The supporting layer and the reagent layer coating film can be better adhered together by means of the resin layer with the adhesion function, so that the requirements on the material and the performance of the base material are reduced, and the cost is saved. The resin layer is a novel resin system which takes water as a dispersion medium instead of an organic solvent, and comprises epoxy resin, alkyd resin, polyester resin, polyurethane resin, modified polybutadiene resin and the like, preferably modified polybutadiene resin; the wet film thickness of the resin layer is 10 to 100 μm, and the dry film thickness is 2 to 50 μm, and the effect is better within 10 to 20 μm. The shell mainly comprises the plastics material, the shell is in the same place through the laminating of supersound laminating mode from top to bottom, be fixed in between the shell from top to bottom with the reagent, it is mainly that the protection reagent is in the transportation, store, the in-process of test is because the deformation that various power leads to, thereby avoid reagent to damage, especially in last quick-witted test, no matter be automatic or semi-automatic send into the detection machine with the dry piece reagent, the dry piece inevitable receives corresponding power, the reagent shell can avoid the reagent dry piece damaged.
The preparation method of the dry tablet reagent comprises the following steps:
1. the resin solution (purchased directly) was uniformly coated on a substrate, dried at 45 ℃ for 10min to form a resin layer having a uniform thickness.
2. And coating the reagent layer solution on the dried resin layer, drying for 30min at the temperature of 45 ℃, and forming a reagent layer after drying.
The formula of the reagent layer solution is as follows:
3. and continuously coating the diffusion layer solution on the dried reagent layer, drying for 5 hours at the temperature of 25 ℃, and finishing the manufacture of the dry powder dry sheet after drying. The formulation of the diffusion layer solution was as follows:
4. cutting the dry powder into a certain size, placing the dry powder into a dry sheet shell matched with a machine, and sealing and storing to finish the manufacture of the dry sheet.
The dry chemical reagent prepared by the method has the advantages of excellent performance, stable reagent, long storage time and convenient carrying, and completely realizes the target of immediate detection.
The dry piece detection operation flow is as follows:
1. taking out the dry slices according to the required amount, and returning the temperature for 1 h;
2. the dried tablets were loaded into the test machine and 10ul of whole blood/plasma/urine/cerebrospinal fluid was added drop wise directly to the sample drop on the upper housing of the dried tablet and tested for 5 minutes to yield a result.
And (5) judging a result: referring to the reference value range, the out-of-range is an abnormal value.
In conclusion, the dry chemical method multi-layer membrane reagent dry tablet for in vitro diagnosis provided by the invention has the advantages of rapid diagnosis, simplicity in operation, cost saving and the like, can improve the accuracy, precision and sensitivity of detection, and has traceability, stable reagent and long storage time.
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.
Claims (9)
1. The utility model provides an albumin quantitative determination dry plate for external diagnosis, including last shell (1) of taking sample dropwise add hole, diffusion layer (2), reagent layer (3), resin layer (4), supporting layer (5), lower shell (6) of taking the light trap, a serial communication port, go up shell (1) and lower shell (6) and laminate each other, it has sample dropwise add hole (7) to go up shell central point to put to open, lower shell central point puts to open has light trap (8), diffusion layer (2), reagent layer (3), resin layer (4), supporting layer (5) from the top down set gradually between last shell (1) and lower shell (6), laminate together in proper order from the top down through the coating process, reagent layer (3) include pigment, buffer solution and hydrophilic polymer.
2. The dry plate for quantitative albumin detection according to claim 1, wherein the pigment is an acid-base indicator capable of binding to albumin to produce a color change, and the acid-base indicator comprises one of bromocresol green, bromocresol purple, bromothymol blue, phenol red, cresol red, cresolphthalein, indigo carmine, methyl red, and methyl orange; the hydrophilic polymer comprises one or more of gel, gelatin derivative, colla Corii Asini, polyvinyl alcohol, polyvinylpyrrolidone, and polyacrylamide; the buffer solution is acetic acid buffer solution or citric acid buffer solution.
3. The dry plate for quantitative measurement of albumin for in vitro diagnosis according to claim 1, wherein the wet film thickness of the reagent layer is 100 to 300 μm, and the dry film thickness is 40 to 200 μm.
4. The albumin quantitative determination dry plate for in vitro diagnosis as claimed in claim 1, wherein the diffusion layer (2) is mainly composed of hydrophilic polymer material and white reflective material, the hydrophilic polymer material is cellulose acetate or carboxymethyl cellulose, the white reflective material is one of titanium dioxide, barium sulfate, polystyrene, microcrystalline cellulose or a mixture of the above reflective materials; the wet film thickness of the diffusion layer (2) is 300-500 mu m, and the dry film thickness is 70-300 mu m.
5. The dry plate for quantitative measurement of albumin as claimed in claim 1, wherein the material of the support layer (5) can be made of various transparent plastics; the thickness of the support layer (5) is 30-200 mu m; the light transmittance of the support layer (5) in the visible light range is more than 85%, and both incident light source and reflected light can pass through the support layer.
6. The dry plate for quantitative detection of albumin for in vitro diagnosis according to claim 5, wherein the transparent plastic is Polyethylene (PE), polypropylene (PP), Polyester Methyl Methacrylate (PMMA), Polystyrene (PS), Polycarbonate (PC) or polyethylene terephthalate (PET).
7. The dry plate for quantitative measurement of albumin for in vitro diagnosis according to claim 1, wherein the resin layer (4) is disposed between the support layer and the reagent layer coating film, the support layer and the reagent layer are adhered together by the resin layer having adhesive effect, and the resin layer (4) is a novel resin system using water instead of organic solvent as a dispersion medium.
8. The dry plate for quantitative measurement of albumin for in vitro diagnosis according to claim 7, wherein the resin layer (4) is made of one of epoxy resin, alkyd resin, polyester resin, polyurethane resin and modified polybutadiene resin; the wet film thickness of the resin layer is 10-100 μm, and the dry film thickness is 2-50 μm.
9. The dry plate for quantitative albumin detection for in vitro diagnosis according to claim 1, wherein the upper housing (1) and the lower housing (6) are mainly made of plastic material, and the upper housing layer and the lower housing layer are attached together by ultrasonic bonding.
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| CN201911056563.1A CN110794147A (en) | 2019-10-31 | 2019-10-31 | Albumin quantitative determination dry tablet for in vitro diagnosis |
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| Application Number | Priority Date | Filing Date | Title |
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| CN201911056563.1A CN110794147A (en) | 2019-10-31 | 2019-10-31 | Albumin quantitative determination dry tablet for in vitro diagnosis |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113125427A (en) * | 2021-04-07 | 2021-07-16 | 中国科学院苏州生物医学工程技术研究所 | Dry chemical reagent sheet for simultaneously detecting uric acid, blood sugar and triglyceride |
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| CN109916893A (en) * | 2019-04-12 | 2019-06-21 | 吉林省汇酉生物技术股份有限公司 | A kind of dry chemistry reagent piece of quantitative detection albumin content and preparation method thereof |
| CN110172496A (en) * | 2019-05-28 | 2019-08-27 | 北京石油化工学院 | A kind of Multilayer film dry plate for surveying hdl concentration |
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2019
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Patent Citations (7)
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| US4042335A (en) * | 1975-07-23 | 1977-08-16 | Eastman Kodak Company | Integral element for analysis of liquids |
| US4732849A (en) * | 1979-11-13 | 1988-03-22 | Fuji Photo Film Co., Ltd. | Multilayered chemical analysis material for analysis of aqueous liquids |
| CN103630535A (en) * | 2013-12-03 | 2014-03-12 | 上海科华生物工程股份有限公司 | Dry chemical test paper for quantitatively determining content of human blood albumin |
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| CN113125427A (en) * | 2021-04-07 | 2021-07-16 | 中国科学院苏州生物医学工程技术研究所 | Dry chemical reagent sheet for simultaneously detecting uric acid, blood sugar and triglyceride |
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Application publication date: 20200214 |