JPH0682444A - Power-free portable incubator - Google Patents

Abstract

PURPOSE:To provide an incubator which can incubate dry elements for analysis regardless of the place of use. CONSTITUTION:A portable incubator for dry elements for analysis comprises a heating surface for heating dry elements for analysis, a storage portion for a heating body provided adjacent the heating surface and making use of exothermic oxidations, and an air flow adjusting portion for adjusting the amount of inlet air for adjustment of the rate of the exothermic oxidations so that the temperature of the heating surface is kept at 30 to 60 deg.C. The incubator requires no power supply. The use of the incubator makes it possible to dry specimen-attached dry elements for analysis under roughly constant conditions regardless of the place of use, thereby providing the possibility of at-home inspection or the like at a high viscosity.

Description

Detailed Description of the Invention

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention supplies a liquid sample to a dry analytical element which does not contain a reagent which directly reacts with a test substance and causes a measurable change, and then transports the analytical element to an analytical institution or the like. The present invention relates to an incubator used for the drying performed before.

[0002]

2. Description of the Related Art Dry analytical elements have been developed mainly for clinical analysis, and are usually used in medical institutions, laboratories, etc. equipped with a reflection optical densitometer. On the other hand, because of the simplicity of the analysis, individual doctors who do not have analytical equipment or home patients, as well as blood tests using it from applicants for screening,
There is a request for urine tests.

As a means for responding to this, the inventors of the present invention used a dry analytical element on which a liquid sample was previously spotted, after stopping and fixing the reaction of the spotted liquid sample, or of the constituent components in the sample. Suppresses deterioration, denaturation, chemical changes, etc., and stores it in a state in which moisture and air are substantially cut off until it is analyzed by an analyzer in a stabilized state. A storage method was developed (Japanese Patent Laid-Open No. 3-28)
9543). In this method, after a liquid sample collected by a patient in daily life at home etc. is spotted on a dry analytical element, it is incubated at a constant temperature for a certain period of time to stop / fix the reaction, or the chemical composition in the sample It is possible to store under constant conditions for a long period of time by making it less likely to be affected by the outside air, especially by the effect of humidity, etc.

[0004]

The above-mentioned preservation method has a problem that the place of use is limited because the incubator requires a power source. An object of the present invention is to provide an incubator capable of incubating a dry analytical element regardless of where it is used.

[0005]

The present invention has been made to achieve the above object, and utilizes a heating surface for heating a dry analytical element and an exothermic oxidation reaction provided adjacent to the heating surface. A portable incubator for a dry analytical element comprising an accommodating portion for a heating element, and an air inflow adjusting portion for adjusting an inflowing amount of air so that a temperature of a heating surface is 30 to 60 ° C. for the rate of the exothermic oxidation reaction, and This object has been achieved by a portable incubator for dry analytical elements that does not require a power source.

The heating element utilizing the exothermic oxidation reaction utilizes the heat generated by the oxidation of iron powder or the like, utilizes the heat of combustion of petroleum, or utilizes the heat of combustion of solid carbon powder such as charcoal or carbon. There are things to do. These can be commercially available products. As the other heating element, one that utilizes heat of neutralization generated by mixing acid and alkali,
Those utilizing the heat of dissolution of calcium chloride, those utilizing the heat generated by mixing an oxidizing agent and a reducing agent, etc. utilizing a chemical reaction other than the above air oxidation reaction,
There are those that use heat storage materials such as hot water, those that utilize heat generated by portable batteries such as dry batteries and solar cells, and the like.

The entire incubator using a heating element utilizing an exothermic oxidation reaction is formed of metal or plastic. It is preferable that the accommodating portion of the heating element is opened at one surface so that the inflow amount of air can be easily adjusted and the other portion is a wall surface through which air cannot pass. If necessary, a small hole for ventilation can be provided in the side wall surface. At least one surface of the heating element accommodating portion may have a structure that can be opened and closed in order to insert and remove the heating element. This opening / closing structure may be opened / closed by a hinge, or may be detachable by fitting. Alternatively, the heating element may be taken in and out by making the air inflow amount adjusting section detachable.

The air inflow control unit limits the inflow of air into the heating element accommodating section to suppress the exothermic reaction rate and prevent the temperature of the heating surface of the dry analytical element from rising too high. This adjustment is done by cloth on the opening surface of the heating element housing,
This can be done by providing a breathable sheet such as paper, a metal plate having holes, a plastic plate, or the like, or by narrowing the opening area itself.

The heating surface of the dry analytical element is provided adjacent to the housing of the heating element. The exothermic reaction rate is adjusted so that the temperature of the heating surface is about 30 to 60 ° C, preferably about 40 to 60 ° C. Since heating is performed to dry the dry analytical element on which the aqueous test sample has been spotted, when the heating element may generate steam like iron powder containing water, the heating surface may not generate steam. The surface that can be blocked.

On the other hand, when the heating element uses something other than the exothermic oxidation reaction, the incubator is a heating surface for heating the dry analytical element, a heating element accommodating portion provided adjacent to the heating surface, and heat generation of the heating element. Is provided so that the temperature of the heating surface is adjusted to 30 to 60 ° C, preferably 40 to 60 ° C.
This adjusting mechanism may be appropriately selected according to the type of heating element and may be provided inside the heating element.

In either type of incubator, a temperature display label or a thermometer can be attached to display the temperature. Also, a timer may be attached to notify the end of the incubation time. Alternatively, the slide or the mainspring may be used so that the slide can be automatically carried out after a predetermined time has passed.

In the dry analytical element, the reagents necessary for the desired analytical reaction are previously incorporated in the dry analytical element,
The product of the reaction is accumulated as the amount of chemical change, and this is based on the principle of determining and quantifying with a visual measuring instrument, etc., but what is applied to the incubator of the present invention, A type that supplies a part or all of the reagent devised in 1) at the time of measurement is preferable.

This is an analytical element containing no measurement reagent in which a liquid sample is supplied, and at least a hydrophilic polymer layer and a microporous layer whose surface is hydrophilized are laminated on a water-impermeable support. In addition, a water-impermeable and gas-permeable layer or a light shielding layer can be provided between the hydrophilic polymer layer and the microporous layer.

The microporous membrane whose surface is hydrophilized may have a blood cell separating ability. The microporous layer having a blood cell separating ability is one whose material and composition are selected so as to specifically separate blood cells and plasma from whole blood without causing hemolysis to the extent that it substantially affects the analytical value. Must be As the microporous layer whose surface is hydrophilized and has blood cell separation ability, acetylcellulose, polyethylene, polypropylene, polystyrene, fluorine-containing polymer, polysulfone, etc. whose surface is hydrophilized can be used.

Examples of the fluorine-containing polymer include poly (vinylidene fluoride) and poly (tetrafluoroethylene), and poly (tetrafluoroethylene) is preferable. The pore size of the microporous layer made of a fluorine-containing polymer is about 0.2.
μm to about 60 μm, preferably about 1 μm to about 20 μm, more preferably 1 to 10 μm, and porosity about 40.
% To about 95%, preferably about 30 μm to about 150 μm, and most preferably about 50 μm to about 120 μm in consideration of handleability such as wrinkling during the manufacturing process.

A microporous layer of a fluorine-containing polymer is described in JP-A-57 / 57
-66359 (US4783315) physical activation treatment (preferably glow discharge treatment or corona discharge treatment) is applied to at least one side of the microporous layer to make the surface of the microporous layer hydrophilic, and to adjoin the adjacent fine particles. The adhesive strength of the adhesive used for partial adhesion with the porous spreading layer can be enhanced.

As the microporous layer of these fluorine-containing polymers, fibrils (fine fibers) of polytetrafluoroethylene described in JP-A-63-501594 (WO 87/02267) can be used.
A microporous matrix layer (microporous layer) consisting of G
ore-Tex (W.L. Gore and Associates), Zite
x (manufactured by Norton), Poreflon (manufactured by Sumitomo Electric Industries, Ltd.) and the like.

Pore size (fine pore size) from about 0.1 μm to about 50
μm, the layer thickness is about 20 μm to about 400 μm. The structure may be unstretched, uniaxially stretched, biaxially stretched, one-layer non-laminated type, two-layer laminated type, for example, a membrane laminated to another membrane structure such as fiber. is there.

In addition, US3368872 (Examples 3 and 4), US3260413 (Examples 3 and 4), JP-A-53-9219
5 (US4201548) and the like, a microporous membrane of polytetrafluoroethylene, and a polyvinylidene fluoride microporous membrane described in US3649505.

Among these microporous membranes of fluorine-containing polymer, those particularly suitable for the microporous layer constituting the blood cell filtration layer have a pore size that is so small as to substantially prevent the passage of red blood cells and a membrane thickness. It is thin and has a high porosity. Specifically, a film having a pore size of 1 to 10 μm, a film thickness of 10 to 200 μm, and a porosity of at least 70% or more is preferable.

The fibril structure or the uniaxially stretched or biaxially stretched non-laminate type microporous membrane is stretched to form a microporous membrane having a large porosity and a short filtration length. Microporous membranes with short filtration length are characterized by high accuracy of quantitative analysis because they are less likely to be clogged by formed components (mainly red blood cells) in blood and the time required for separating blood cells and plasma is short. .

In forming the microporous membrane of these fluorine-containing polymers, one kind or two or more kinds of fluorine-containing polymers may be mixed, or one kind or two or more kinds of polymers or fibers containing no fluorine may be mixed. It may be mixed with and formed into a film.

In order to impart sufficient hydrophilicity to the microporous membrane of a fluorine-containing polymer so that the aqueous liquid sample can be diffused, permeated and transferred to the surface or inside of the membrane without being repelled, it is generally necessary to use a fine fluorine-containing polymer. About 0.01% to about 10%, preferably about 0.1% to about 5.0%, more preferably 0.1% to 1%, of the void volume of the porous membrane coats the surface of the voids of the microporous membrane. It is necessary. For example, the thickness is 50μ
In the case of the microporous membrane of m fluorine-containing polymer, the amount of surfactant to be impregnated is preferably generally in the range of 0.05 g / m ² of 2.5 g / m ^2. As a method of impregnating a microporous film of a fluorine-containing polymer with a surfactant, an organic solvent (eg, alcohol, ester, ketone) having a low boiling point of the surfactant (preferably having a boiling point of about 50 ° C to about 120 ° C) is used. )
After soaking the microporous membrane of the fluorine-containing polymer in the solution and allowing the solution to penetrate the internal voids of the microporous membrane substantially sufficiently, the microporous membrane is gently pulled up from the solution and blown (warm air The preferred method is to feed (dry) and dry. A surfactant may be contained in the microporous membrane of the fluorine-containing polymer together with the components such as the pretreatment reagent contained in the microporous layer constituting the blood cell filtration layer.

Surfactants used for hydrophilizing a fluorine-containing polymer microporous membrane include nonionic (nonionic), anionic (anionic), cationic (cationic), amphoteric Any surfactant can be used. Among these surfactants, the nonionic surfactant has a relatively low effect of hemolyzing erythrocytes, and is therefore advantageous in a multilayer analytical element for analyzing whole blood. As the nonionic surfactant, alkylphenoxy polyethoxy ethanol, alkyl polyether alcohol, polyethylene glycol monoester, polyethylene glycol diester, higher alcohol ethylene oxide adduct (condensate), polyhydric alcohol ester ethylene oxide adduct (condensate), There are higher fatty acid alkanolamides.

The following are specific examples of the nonionic surfactant. Examples of the alkylphenoxypolyethoxyethanol include: isooctylphenoxypolyethoxyethanol: (Triton X-100: containing oxyethylene units on average 9 to 10) (Triton X-45: containing oxyethylene units on average 5) nonylphenoxypolyethoxyethanol: ( IGEPAL CO-630: Oxyethylene unit average 9
Contained) (IGEPAL CO-710: Oxyethylene unit average 10
(Containing 11 to 11) (LENEX 698: containing 9 of oxyethylene units on average) As the alkyl polyether alcohol, a higher alcohol polyoxyethylene ether: (Triton X-67: CA Registry No.59030-15-
8)

The fluorine-containing polymer microporous membrane may be hydrophilized by providing one or more water-insoluble water-soluble polymers in its porous space. As examples of water-soluble polymers, polyvinyl alcohol, polyethylene oxide, polyethylene glycol, methyl cellulose, ethyl cellulose, hydroxyethyl cellulose, hydroxypropyl cellulose for hydrocarbons containing oxygen, polyacrylamide, polyvinyl pyrrolidone, polyvinyl amine for those containing nitrogen, Polyethyleneimine and those having a negative charge include polyacrylic acid, polymethacrylic acid, polystyrene sulfonic acid and the like. The insolubilization may be performed by heat treatment, acetalization treatment, esterification treatment, chemical reaction with potassium dichromate, crosslinking reaction with ionizing radiation, or the like. For details, see JP-B-56-2094 and JP-B-56-1618.
No. 7 publication.

The polysulfone microporous membrane is prepared by dissolving polysulfone in dioxane, tetrahydrofuran, dimethylformamide, dimethylacetamide, N-methyl-2-pyrrolidone, a mixed solvent thereof or the like to prepare a membrane-forming stock solution, which is supported. It can be produced by casting on the body or directly in a coagulating liquid, washing and drying. Details are disclosed in JP-A-62-27006.
A microporous membrane of polysulfone is also disclosed in JP-A-56-1264.
No. 0, JP-A-56-86941, JP-A-56-154051 and the like are also disclosed, and these can also be used. The microporous membrane of polysulfone can also be hydrophilized by containing a surfactant like the fluorine-containing polymer or by providing a water-insoluble water-soluble polymer.

A porous spreading layer can be provided between the microporous layer having a blood cell separating ability and the hydrophilic polymer layer.
Between the porous development layer and the microporous layer having hydrophilicity on the surface and having the ability to separate blood cells, it can be peeled after supplying the liquid sample even in the state where it is not substantially peeled off, so-called part described later. It may be in a bonded state.

The porous spreading layer has a function of spreading the components contained in the aqueous sample in a plane without being substantially unevenly distributed and supplying the hydrophilic polymer layer at a substantially constant amount per unit area. Any known non-fibrous and fibrous porous materials can be used as the spreading layer, which is a layer that has been used in the dry chemistry analysis element.

The spreading layer may contain a nonionic, anionic, cationic or amphoteric surfactant in order to accelerate the spreading of the sample. Further, a development control agent such as a hydrophilic polymer may be included for the purpose of controlling the development property. Further, various reagents for accelerating the target detection reaction, or for reducing or preventing interference or interference reactions, or a part of the reagents can be included. The thickness of the spreading layer is 20 to 200 μm, preferably 50 to 1
The thickness is 70 μm, more preferably 80 to 150 μm.

The hydrophilic polymer layer is soluble in known water used in dry chemistry analysis elements,
Various swelling and hydrophilic polymers can be used.
A natural or synthetic hydrophilic polymer having a swelling ratio upon absorption of water at 30 ° C. of about 150% to about 2000%, preferably about 250% to about 1500% can be used. Akira
59-171864, 60-108753 and the like (eg, acid-treated gelatin, deionized gelatin, etc.), gelatin derivatives (eg, phthalated gelatin, hydroxyacrylate graft gelatin, etc.), agarose, pullulan, pullulan derivatives, Examples thereof include polyacrylamide, polyvinyl alcohol, polyvinylpyrrolidone, etc., but are not limited thereto. Instead of the hydrophilic polymer layer, paper having a hydrophilic surface or a polymer porous membrane can be used.

The thickness of the hydrophilic polymer layer is about 1 when dried.
μm to about 100 μm, preferably about 3 μm to about 50 μm, particularly preferably about 5 μm to about 30 μm, and preferably substantially transparent. The hydrophilic polymer layer may contain various reagents or a part of the reagents for promoting the intended reaction or for preventing or reducing the interference or interfering reaction.

As the water-impermeable support, known water-impermeable supports conventionally used in dry chemistry analysis elements can be used. Specifically, polyethylene terephthalate, bisphenol A polycarbonate, polystyrene, cellulose ester (for example,
Sululose diacetate, cellulose triacetate,
A transparent film made of cellulose acetate propionate or the like) having a thickness of about 50 μm to 1 mm, preferably about 80 μm to about 300 μm can be used.

The support is usually a light-transmissive one, but when it is measured from the spreading layer side, it may be colored or light-impermeable. If necessary, a publicly known undercoat layer or an adhesive layer may be provided on the surface of the support to strengthen the adhesion with the hydrophilic polymer layer.

Here, one of the features of the dry analytical element of the type in which the reagent is supplied later is a partial adhesion between the microporous layer made of a fluorine-containing polymer and the surfactant and the porous spreading layer (porosity). Adhesive bonding) will be described.

Partial adhesion refers to JP-A-61-4959 (EP0166).
365A), JP-A-62-138756-138758 (EP0226465A)
And the like, wherein the two adjacent porous layers are adjacent to each other or the adjacent porous layer and the non-porous layer are adhered to each other, and "partially (or intermittently) between the interfaces of the two adjacent layers". The adhesive is substantially adhered to and integrated with the adhesive disposed in the above, and is configured so that the uniform passage of the liquid is not substantially obstructed between the two adjacent surfaces.

In the analytical element used in the present invention, the interface between the microporous layer made of a fluorine-containing polymer and the porous spreading layer may be partially bonded (porous bonded). It is a general partial adhesion method that an adhesive is partially disposed on the porous spreading layer, and then the microporous layer made of a fluorine-containing polymer is bonded while uniformly applying light pressure. On the contrary, it is also possible to partially dispose the adhesive on the microporous layer made of a fluorine-containing polymer, and then to bond the porous spreading layer while applying light pressure uniformly. Further, an adhesive is partially disposed on the microporous sheet material to be the porous development layer, and the microporous layer made of a fluorine-containing polymer is evenly and pressure-bonded to it, or vice versa. Partially placing the adhesive on the microporous layer made of a fluorine-containing polymer,
Then, after laminating the microporous sheet material to be the porous development layer uniformly and lightly, apply the microporous sheet material to be the development layer uniformly to the hydrophilic polymer layer. You can also

A method of partially disposing an adhesive in a microporous layer or a porous spreading layer made of a fluorine-containing polymer is disclosed in JP-A-2006-29187.
61-4959, JP-A-62-138756, JP-A-64-23160 (DE37
21236A) and the like.
Of these methods, the printing method is preferred. Among the printing methods, a method of transferring an adhesive to a porous layer or a detection functional layer using a printing plate (a gravure printing plate or an intaglio is preferable) roller to attach the adhesive and a method of bonding two adjacent layers are, for example, It can be carried out by a known device and method described in "Printing Engineering Handbook" edited by Japan Society of Printing Science (Gihodo Publishing Co., Ltd., 1983), pages 839 to 853 and the like. As the adhesive used, various kinds of adhesives described in JP-A-62-138756 and other known adhesives described in the above-mentioned "Printing Engineering Handbook", pages 839 to 853 and the like can be used.

Furthermore, in the embodiment in which the microporous layer made of the fluorine-containing polymer is peeled off after spotting the sample, the spreading layer and the microporous layer do not need to be adhered to each other, and the diffusion / permeation of the sample does not occur. It suffices that they are laminated so as to proceed quantitatively.

Next, the test substance will be described. In the present invention, the target test substance is not particularly limited. In addition to enzymes, lipids, inorganic ions, metabolites, proteins, etc., which are usually measured in the field of clinical testing, various globulin, immune antigens, biologically-derived components such as immune antibodies, drugs, hormones, etc.
As long as an analysis method such as a tumor marker is established, it can be an analysis target.

The dry analytical element can take various configurations described below depending on the item or sample to be measured. 3 and 4 show basic configuration examples.

1: Analytical element having a structure comprising a water-impermeable support / hydrophilic polymer layer / developing layer / fluorine-containing polymer and a microporous layer containing a surfactant. Ca, GOT (Glutamate oxaloacetate transaminase), GPT
(Glutamate pyruvate transaminase), γ-
GTP (γ-glutamyl transpeptidase), glucose, LDH (lactate dehydrogenase), CPK (creatine phosphokinase), TP (total protein), Alb (albumin), TCHO (total cholesterol), UA (uric acid), medium It is effective for analyzing sex fat.

2: A water-impermeable support / hydrophilic polymer layer / developing layer / microporous layer containing a surfactant and containing a surfactant, wherein the hydrophilic polymer layer and / or the developing layer contain Analytical elements that include chromogens. As a chromogen,
Ann. Clin. Biochem., 6, 24-27 (1969), 4-aminoantipyrine (also known as 4-aminophenazone,
That is, 1-phenyl-2,3-dimethyl-4-amino-3-pyrazolin-5-one), and 1- (2,4,6-trichlorophenyl) -2,3 described in JP-A-59-54962.
-Dimethyl-4-amino-3-pyrazolin-5-one,
1- (3,5-dichlorophenyl) -2,3-dimethyl-
Tri-substitution such as 4-amino-3-pyrazolin-5-one-
4-Amino-3-pyrazolin-5-one, Japanese Patent Publication 55-25
4-aminoantipyrine analogs such as 1-phenyl-2,3-dimethyl-4-dimethylamino-3-pyrazolin-5-one described in 840 etc. can be used. Among these compounds, 4-aminoantipyrine, 1-
(2,4,6-trichlorophenyl) -2,3-dimethyl-4-amino-3-pyrazolin-5-one, 1- (3,
5-dichlorophenyl) -2,3-dimethyl-4-amino-3-pyrazolin-5-one and the like are preferable.

3: A water-impermeable support / hydrophilic polymer layer / developing layer / fluorine-containing polymer in the constitution of a microporous layer containing a surfactant, wherein the hydrophilic polymer layer and / or the developing layer contain , Analytical elements including chromogens and other reagents (excluding measurement reagents described later). Other reagents include POD (peroxidase), NAD (nicotinamide adenine dinucleotide), NADP (nicotinamide adenine dinucleotide phosphate), DIP
(Diaphorase) and the like.

In the configurations 2 and 3, the chromogen or other reagents can be supplied after the liquid sample is supplied and stabilized, but since most of the chromogens are water-insoluble, they are separated from the measurement reagent. It is necessary to supply the chromogen and other reagents, and it is advantageous to reproducibly produce the layer by including it in the layer from the beginning.

4: Analytical element containing mordant layer. When the coloring reagent forms an ionic dye, a mordant layer can be provided between the water-impermeable support and the reagent layer. The efficiency of optical detection can be improved by transferring and trapping the dye generated in proportion to the amount of the test substance in the sample to the mordant layer.

For example, when the coloring dye forms a cationic dye, a hydrophilic polymer layer containing a polymer containing an anion atom or an atomic group bonded to a polymer chain is used as a mordant layer, and a coloring reagent. When forming an anionic dye, a hydrophilic polymer layer containing a polymer containing a cation atom or an atomic group bonded to a polymer chain can be used as the mordant layer. For details of these mordant polymers, see Japanese Examined Patent Publication No. 2-30466 and JP-A-51-4.
0191, 54-29700, 53-131089 and the like.

For example, as an anion mordant polymer,
Japanese Patent Publication No. 2-30466, column 13 to column 14, an alkaline hydrolyzate of a methyl vinyl ether-maleic anhydride copolymer, an alkali metal salt or an alkaline earth metal salt of polystyrene-p-sulfonic acid, Styrene-p
-Alkali metal salts or alkaline earth metal salts of copolymers of sulfonic acid and hydrophilic vinyl monomers, and the like. Further, layers and the like capable of containing these polymers are also described in detail in columns 15 to 16 of the publication.

5: Analytical element in which the light shielding layer is provided between the hydrophilic polymer layer and the spreading layer in the constitutions 1 to 4 above.
Whole blood can be used as a sample without removing the microporous layer made of a fluorine-containing polymer and containing a surfactant.
The light-shielding layer is a water-permeable material in which fine particles or fine powders having light-shielding properties or both light-shielding properties and light-reflecting properties (hereinafter simply referred to as “fine particles”) are dispersed and held in a hydrophilic polymer binder having a small film-forming ability. Or a water-permeable layer. The light shielding layer shields the color of the supplied aqueous liquid sample, especially the red color of hemoglobin contained in the whole blood sample, when the detectable change (color change, color development, etc.) is reflected and measured from the light-transmissive support side. In addition, it also functions as a light reflection layer or a background layer.

Examples of fine particles having both light shielding property and light reflecting property are titanium dioxide fine particles (rutile type, anatase type or brookite type fine crystal particles having a particle size of about 0.1 μm to about 1.2 μm, etc.), barium sulfate fine particles. , Aluminum fine particles or fine flakes, and examples of light-shielding fine particles include carbon black, gas black, carbon microbeads, etc. Among these, titanium dioxide fine particles,
Barium sulfate fine particles are preferred.

As the hydrophilic polymer binder having a film-forming ability, there are weakly hydrophilic regenerated cellulose, cellulose acetate and the like in addition to the above-mentioned hydrophilic polymers. Among these, gelatin, gelatin derivatives, polyvinyl alcohol, polyacrylamide, Maleic acid copolymers and the like are preferred. A known hardening agent (crosslinking agent) can be mixed and used for gelatin and a gelatin derivative.

6: An analytical element in the above constitutions 1 to 4, in which a water-impermeable and gas-permeable layer (hereinafter referred to as a barrier layer) is provided between the hydrophilic polymer layer and the spreading layer. It is effective for analysis of BUN (urea nitrogen), CRE (creatinine), CO _{2 and the} like which generate ammonia gas by the reaction. Either whole blood or plasma can be used as the sample. As the barrier layer, the uniform polymer coating layer disclosed in JP-A-52-3488, the membrane filter disclosed in JP-A-58-77661 and the like can be used.

7: Analytical element having a structure obtained by removing the development layer from the structures 1 to 6 above. That is, an element that also utilizes the microporous layer as a spreading layer. In all of the above elements, the microporous layer may be substantially fixed to the adjacent layer, or may be in a peelable state, especially when whole blood is used as the sample,
Except for the structure having a light shielding layer, it is preferably in a peelable state.

The measuring reagent refers to a reagent which directly reacts with a test substance to be analyzed to cause a chemical change. That is,
When the enzyme is a test substance, its substrate is; when the test substance is an antigen (antibody), it is an antibody (antigen); and the test substance is a lipid, sugar, or metabolite, which can be detected by the enzyme When it is a compound that causes a significant change, it is the enzyme.
In addition, when these reactions are caused by a general chemical reaction by a chemical reagent other than an enzyme, it refers to a corresponding chemical substance. A specific example will be described below.

When the test substance is GOT which is an enzyme,
Its substrates are aspartic acid and α-ketoglutaric acid, high-molecular-weight starch or low-molecular-weight oligosaccharides for amylase, L-γ-glutamylparanitroanilide for GGT, and para-nitrophenyl phosphate for ALP. is there.

Further, glucose is glucose oxidase, uric acid is uricase, cholesterol is cholesterol esterase or cholesterol oxidase, neutral fat is lipase or esterase, and urea is urease.

When an analyte is a protein, albumin, Ca, inorganic phosphorus or the like, and a test substance directly reacts with an indicator or the like to cause a detectable change, the term "indicator" is used. One of the purposes of supplying this reagent later is to prevent deterioration of the detection reagent that occurs during storage of the analytical element, which is a drawback of conventional dry chemistry, so that the reaction incorporated in the above reaction system is not performed. When the reagent is unstable such as an enzyme, it is preferable that these are also contained in the measurement reagent.

That is, regarding the distribution of the reagent to be contained in the measurement reagent solution and the reagent to be contained in the analytical element,
It can be variously changed by using the analytical performance and storage stability as an index. Of course, even if there is only one analysis target, the above distribution differs depending on the assembly of the detection reaction system.

Among the measuring reagents, in order to proceed the reaction stably and with good reproducibility, pH and ionic strength are adjusted, diffusion and permeation into the material constituting the analytical element are improved, and an enzyme contained therein is included. Various reagents may be included for the purpose of improving instability.

Further, the measurement reagent may contain a reagent for inhibiting a reaction that competes with the detection reaction.
Examples of such a reagent include bilirubin oxidase and ascorbate oxidase. Further, for isozyme detection, a compound that inhibits an enzyme derived from a specific organism, such as an inhibitor of P-type amylase, can be included. Furthermore, in whole blood measurement, NaN ₃ or the like which is effective as an inhibitor of the hemoglobin catalase activity can be added.

The dry analytical element using the incubator of the present invention is not limited to the analytical element of the type in which a part or all of the above-mentioned reagent is supplied later, and it is a known element containing a reagent in advance. Good.

Examples of such a dry analytical element include a test paper and a multilayer chemical analysis film. As the test paper, various types such as those for whole blood, plasma / serum, and urine can be used, and types for diluting a sample can also be used. Various types of multilayer chemical analysis films are known, and any of them can be used.

As a method of using the incubator of the present invention, a dry analytical element on which a predetermined amount of a sample to be tested is spotted and developed is placed on a heating surface, and a heating element is caused to generate heat to be contained in the sample to be tested. It is sufficient to evaporate the water contained therein. Heating temperature is 30
~ 60 ℃, preferably 40 ~ 50 ℃, heating time 1-10
Minutes are enough.

Here, the term "dry" means that the reaction does not substantially proceed in the hydrophilic polymer or the deterioration of the test substance does not proceed. Therefore, depending on the analysis target, for example, when targeting an enzyme, the residual water content in the analytical element is the amount of water supplied as the sample.
It may be 50%, preferably 10% or less, more preferably 1% or less.

The analytical element that has been dried by heating is sent to an inspection agency in a state where it is kept in a dry state in a temperature-proof container or the like, and is analyzed there. As a means for maintaining this dry state, for example, the method described in JP-A-3-289543 can be used. For example, there is a method of sealing in a metal box provided with a water removing means or a bag made of a film or sheet of an organic polymer or metal that does not allow water to permeate.

As the means for removing water, any known hygroscopic agent that does not substantially change the quality of the sample can be appropriately selected and enclosed. After bagging the element, it may be aired out and squeezed out.

In the case of an analytical element of the type in which a reagent is supplied later, analysis is carried out by the following method. The measurement reagent solution corresponding to the item to be analyzed is supplied to the analysis element taken out from the closed container to cause a reaction. This reaction is carried out by a method known in the field of dry chemistry (reflection density photometry,
Color change, fluorescence measurement, luminescence measurement, etc.) to quantify the components contained in the sample. If necessary, the microporous layer made of a fluorine-containing polymer and containing a surfactant may be removed.

As the measuring reagent solution corresponding to the item to be analyzed, a known reagent solution in wet chemistry can be used. These react with the analyte and
A change that can be detected mainly by an optical measurement method, for example, a color change, color development (coloring), fluorescence, light emission, a change in absorption wavelength in the ultraviolet region, a change in turbidity, or the like occurs.

As a method for measuring dry chemistry, a reflective optical system is usually used. Also in the method of the present invention,
The method of photometry through the water-impermeable support of the analytical element has the widest range of application, but when the sample is not whole blood or when the measurement is performed by removing the microporous layer after supplying the sample, the transmission photometric method is used. Can be measured by When the water-impermeable support is opaque, it can be measured from the opposite side of the support.

FIG. 5 shows a schematic diagram of the measuring method according to the present invention.

[0071]

[Function] In clinical analysis by dry chemistry, particularly dry chemistry of the type in which reagents are supplied later, if incubation (warming) conditions are within a certain range, for example, when temperature control is strictly performed at ± ° C. The present inventor has found that the same level of accuracy and precision can be realized.

However, since the electric type incubator has been used so far, there are great restrictions on home inspection and field inspection. The feature of dry chemistry of the type that supplies reagents later is that if a small amount of whole blood is spotted on a chemical slide even at home or outdoors (blood donation, mass examination, medical depopulated area), even if dehydrated and dried The chemical component is that it can be stored safely. Electrothermal heating is the most accurate method for dehydration and drying, but it cannot be used in the absence of a power source. The present invention has solved this problem.

[0073]

【Example】

Example 1 (1) Preparation of incubator The incubator shown in FIGS. 1 and 2 was prepared. The incubator is made of a plastic plate with a thickness of about 0.5 mm and has a box shape of 50 mm × 100 mm × 20 mm. Inside the box, 10 mm below the upper edge, an air inflow control section 1 is formed by stacking two stainless steel net plates having an opening area of about 1 to 50%. The opening area and number of the mesh plate are defined by the type and amount of the heating element, the amount of the analytical material to be processed, the amount of the sample liquid, and the like. The lower part of the air inflow control unit 1 is the heating element 2
It is the accommodation section 3 of The bottom surface 4 of the box has a hinge lid structure, and the heating element 2 can be taken in and out by opening and closing the hinge lid structure. In this incubator, the upper surface of the air inflow control section 1 is used as a heating surface 5, and a dry analytical element heating container 6 made of stainless steel is placed thereon. This heating container 6 has exactly 2 dry analysis elements 7.
They are sized so that they can be placed side by side, and have a box shape with a side wall height of 10 mm so that water vapor generated from the heating element 3 can be avoided.

(2) Preparation of Analytical Element 0.2% of nonionic surfactant p-nonylphenoxypolyglycidol (containing an average of 10 glycidol units) on a transparent polyethylene terephthalate film base having a thickness of 180 μm and having a base coat for gelatin. Containing gelatin,
It was applied and dried so that the dry film thickness was about 15 μm.
A polyester knitted fabric hydrophilically treated with a surfactant was laminated thereon according to the method described in JP-A-62-224299. Further, 7% of polyvinylpyrrolidone (average molecular weight: 1.2 million) was applied on the polyester knit development layer so that the application amount was 2 g / m ² .

(3) Preparation of glucose measuring reagent A glucose measuring reagent was prepared by dissolving the following reagent in 10 ml of 0.1 M MES (2- (N-morpholino) ethanesulfonic acid) buffer solution having a pH of 5.7. GOD 1300U POD 5000U Dihydrooxynaphthalene 110mg 4-Aminoantipyrine 140mg p-Nonylphenoxypolyglycidol 800mg

(4) Preparation of Specimen Control serum and glucose were added thereto to give a glucose concentration of 87 mg / dl, 179 mg / dl and 283
A total of 4 types of samples of mg / dl were prepared.

(5) Measurement of glucose concentration The heating element 2 (Hokaron, manufactured by Lotte Co., Ltd.) utilizing the exothermic reaction due to the oxidation of iron powder was placed in the incubator of (1) above and left for 5 minutes. However, the temperature of the heating surface 5 is 50 ° C.
Became. 10 μl of each sample was spotted on the dry analytical element of the above (4), which was housed in the heating container 6 and placed on the heating surface 5 of the incubator and left for 5 minutes. The temperature change during that period is 32 ~
It was 41 ° C. Then, set each analytical element in a reflection optical densitometer (“Fuji Drychem Analyzer 5500”, manufactured by Fuji Photo Film Co., Ltd.), and suck the measurement reagent solution prepared in (5) into the dispenser for normal measurement. Spotting, incubation, and photometry were performed according to the operation. The sample spotting, dehydration drying, and measurement were repeated using 10 analytical elements in the same manner as above, and the reproducibility of the measurement was examined. The results were as follows. Glucose concentration level in sample L MH Average of 10 measurements (mg / dl) 87 179 283 SD (mg / dl) 4.1 4.3 8.8 CV (%) 4.7 2.4 3.1

Example 2 (1) Preparation of incubator Inside a 50 mm × 100 mm × 20 mm stainless steel box, a heating element utilizing the exothermic reaction due to oxidation of iron powder (“Hokaron / shoes”, Lotte Co., Ltd.) (Manufactured by the company) was placed between two upper and lower duck cloths to form an incubator.

(2) Preparation of Analytical Element 0.2% of nonionic surfactant p-nonylphenoxypolyglycidol (containing an average of 10 glycidol units) on a 180 μm thick polyethylene terephthalate transparent film base with a subbing for gelatin. Containing gelatin,
It was applied and dried so that the dry film thickness was about 15 μm.
A polyester knitted fabric hydrophilically treated with a surfactant was laminated thereon according to the method described in JP-A-62-224299. Further, 7% of polyvinylpyrrolidone (average molecular weight: 1.2 million) was applied on the polyester knit development layer so that the application amount was 2 g / m ² .

(3) Hydrophilization treatment of fluorine-containing polymer Polytetrafluoroethylene microporous film, Zitex A1
35 (manufactured by Norton, USA) was impregnated with an ethanol solution containing 20% of Triton X-100 to remove air contained in the micropores to allow the activator solution to sufficiently penetrate into the micropores. After that, it was pulled out from the solution and dried by blowing dry air at 50 ° C.

(4) The analytical element prepared in (2) is set to 15 mm × 15 mm
It was cut into the size of. On the other hand, the hydrophilized fluorine-containing polymer microporous membrane prepared in (3) was cut out into a size of 15 mm × 15 mm, and it was placed on the analysis layer. This is described in JP-A-57-
The analytical element was completed by incorporating it into a plastic frame by the method described in 63452.

(5) Preparation of Amylase Assay Reagent Solution 10 ml of HEPES buffer (0.1 M) solution having pH 7.3 was added with α
-Glucosidase 6000 U and 540 mg of para-nitrophenyl oligosaccharide (having a glucose number of 5) were dissolved to prepare an amylase measurement reagent solution.

(6) Spot application of sample 30 μl of whole blood sample obtained by collecting heparin from a healthy person was spotted on the analytical element prepared in (4) above using a micropipette. Whole blood spread laterally on the microporous membrane and penetrated inside. After leaving for 1 minute, the microporous membrane was peeled off and removed, and the resultant was housed in the same heating container as in Example 1, placed on a Zuk cloth laid on the heating element of the incubator of (1), and dried by heating for 5 minutes. I did. The temperature change during that time
It was 31-43 ° C. The slide after the drying treatment was set in an analyzer in the same manner as in Example 1, and the amylase activity was measured. However, the measurement wavelength was 400 nm and the photometric time was 5 minutes-1 minute. The same operation was repeated 10 times and the simultaneous reproducibility was examined.
The results are shown below and showed good quantification. Average value 18U / L S D 3.1U / L C V 17%

[0084]

EFFECTS OF THE INVENTION By using the incubator of the present invention, it is possible to dry a dry analytical element on which a sample is spotted under almost constant conditions regardless of the place of use, and open up a way to perform home inspection and the like with high viscosity. It is a thing.

[Brief description of drawings]

FIG. 1 is a sectional view showing a usage state of an incubator which is an embodiment of the present invention.

FIG. 2 is a perspective view of the above usage state.

FIG. 3 is a schematic diagram showing an example of the basic constitution of a dry analytical element containing no measurement reagent, which is used in the measuring method of the present invention.

FIG. 4 is a schematic diagram showing another basic configuration example of a dry analytical element containing no measurement reagent, which is used in the measuring method of the present invention.

FIG. 5 is a schematic diagram showing a measuring method of the present invention.

Claims (2)

[Claims] 1. A heating surface for heating a dry analytical element, a heating element accommodating portion provided adjacent to the heating surface for utilizing an exothermic oxidation reaction, and a rate of the exothermic oxidation reaction to a temperature of the heating surface. Portable incubator for dry analytical elements, which consists of an air inflow control unit that regulates the air inflow so that the temperature is between 30 and 60 ℃ 2. A portable incubator for a dry analytical element that does not require a power source.