JPH0682445A - Analyzer for dry analysis element - Google Patents

Abstract

PURPOSE:To analyze desired constituents including specific items in a dry analysis method by providing a quantitative supply device for a sample liquid and that for a reagent. CONSTITUTION:Constant volume-supply devices 1, 2 for a sample liquid and a reagent have turn tables 3, 4 and dispensation arms 5, 6. Cups 7 for the sample liquid or reagent and quantitative constant volume-supply microchips 8 are alternately disposed. The arms 5, 6 are rotated and lowered and cause each microchip 8 to fit with a mounting rod 9, and slightly rotate the tables 3, 4 after being raised, so as to retain a certain amount of sample liquid or reagent in each microchip 8. Next, the arms 5, 6 are moved right above an analytical slide 10 and a drop of sample liquid or reagent is attached to the slide 10. For items that generally require analysis, all of elements for analysis in which the reagent is set are used, while for specific items the elements for analysis which do not contain part or the whole of the reagent are used; the other reagent is supplied as drops during measurement.

Description

Detailed Description of the Invention

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a dry analytical element analyzer and a dry analytical element analyzer capable of measuring an analytical element containing all reagents necessary for analyzing a target component and an analytical element not containing at least a part thereof. It relates to the measurement method that was used.

[0002]

2. Description of the Related Art Dry analysis uses a dry analysis element in which all reagents necessary for analyzing a target component are incorporated into a filter paper or a multi-layer material in which one or more water-permeable layers are laminated on a support. Since it is an analytical method that is performed in a simple manner and can obtain high analytical accuracy while being easy to operate, it has been actively developed recently.

[0003]

Although the analytical element containing all of these reagents is convenient, it is costly due to small-scale production when trying to meet the analytical requirements of special items specific to the facility or market segment. It was difficult to put it into practical use. An object of the present invention is to provide a practicable analyzer capable of analyzing various target components including special items required by each facility or market by a dry analysis method, and a measuring method using the analyzer.

[0004]

Means for Solving the Problems The present inventor has made diligent studies in order to achieve the above-mentioned object and devised an analytical element which does not contain a part or all of the reagents necessary for the analysis. In order to analyze various analytical items required by using, a large number of measurement reagents had to be prepared and managed, which was not practical. Therefore, we proceeded with further study, and for the items that are requested to be analyzed universally, we used the analytical elements that incorporate all reagents, and for the special items, we analyzed them by using the analytical elements that do not include some or all of the reagents. The inventors have devised a method for spot-supplying reagents not included in the measurement, and have found that this method can achieve the above object, and have completed the present invention.

That is, the present invention relates to a dry analytical element analyzer characterized by having at least a fixed amount supply device for a sample liquid and a fixed amount supply device for a reagent liquid, and a measuring method using the same.

The dry analytical element analyzer comprises an incubation section for advancing the reaction while keeping the analytical element at a constant temperature for a predetermined time, a photometric section for measuring an optical change caused by the reaction, and a display section for displaying the photometric result. And at least a fixed amount supply device for sample liquid,
A supply device, a transfer device, etc. for the analysis slide containing the analysis element are appropriately provided.

The incubation section is for heating the analytical element on which the sample or the like is spotted, for example, for 1 to 15 minutes in a range of about 30 to 60 ° C. under constant conditions, and the photometric section is for the optical change caused by the reaction. Is usually measured using a reflection optical densitometer. The display unit may be a meter unit of a reflection optical densitometer, but recently, a type in which a computer having a built-in calibration curve performs an arithmetic operation and directly displays the concentration digitally or prints out is increasing. . Various types of such analyzers for dry analytical elements are commercially available (for example, “Fuji Drychem 1000”, Fuji Photo Film Co., Ltd.).
Products, etc.), "Medical Electronics and Biotechnology" (Japan M.
E Academic Society, Vol. 22, No. 4, pp. 274-278, 1984).

In the analyzer of the present invention, at least two liquid quantitative supply devices, that is, a reagent liquid quantitative supply device and a sample liquid quantitative supply device are additionally provided to these known analyzers. However, there is a feature. For both quantitative supply devices, commercially available devices may be used. For example, the storage parts for the samples, reagents, etc. are arranged in a circle along the periphery of the turntable, and the liquid stored therein is attached to the tip of the rotating arm. A fixed amount of liquid may be sucked from the liquid storage parts arranged in the vertical direction or the horizontal direction by aspirating a fixed amount with a pipette or the like and rotating and spotting this on the analysis slide. Alternatively, the liquid may be spotted on the analysis slide by moving the analysis slide right above the analysis slide in a direction perpendicular to the running direction of the analysis slide and descending. It goes without saying that all spots are placed upstream of the incubator. Two or more fixed-quantity supply devices for reagent liquid can be provided.

The analyzer of the present invention analyzes a plurality of types of analytical elements, that is, at least one analytical element in which all reagents are incorporated and at least one analytical element that does not include some or all of the reagents. Therefore, it is preferable to provide a mechanism for identifying whether at least the analysis element contains all reagents and only the sample needs to be spotted, or whether the reagent needs to be spotted further. . When it is necessary to change the amount of the sample to be spotted or the type of reagent to be spotted is changed according to the analysis item, it is preferable to be able to identify the analysis item. Various means can be used as the identification mechanism, but a method of printing a barcode on a slide frame is simple (for example, Japanese Patent Laid-Open No. 63-172944).

In the dry analytical element applied to the analyzer of the present invention, the reagents necessary for the intended analytical reaction are previously incorporated into the dry analytical element, and the reaction product is accumulated as a chemical change amount. However, there are those based on the principle of determining and quantifying this with a visual measuring instrument and those lacking some or all of the above reagents. It should be noted that chemical substances such as hemoglobin amount and bilirubin in which the test substance itself can be detected do not require chemical reaction. The detection method may be any method such as fluorescence, luminescence, magnetism, etc., in addition to the optical density measurement. Further, after fixing once, another reagent can be added to make the detection more accurate and reliable. As such a dry analysis element, for example,
There are test strips and multilayer chemical analysis films. 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.

The dry analytical element has at least one liquid-permeable porous layer. This layer may contain all or some of the reagents that react with the component to be detected. A part of the reagent may be contained in each layer of different depths of one porous layer. Another non-porous liquid permeable layer may contain a part or all of the above reagents. It may have more than one porous or non-porous liquid permeable layer.

The dry analytical element comprises at least one interactive composition. The interaction composition is one or more activities that interact with the analyte or a reaction product or degradation product of the analyte, or with each other, when a liquid sample containing the analyte is contacted with the dry analytical element. Including ingredients. In the dry analytical element, such an interaction produces, decomposes or inactivates a dye, a fluorescent substance or the like which can be detected directly or indirectly (via another reaction) by spectrophotometry. A pigment may be produced by the interaction of a pigment-producing substance with a required specific component in blood,
It may also be produced by releasing the diffusible dye from the non-diffusible dye. "Interaction" includes chemical activity, catalytic activity such as in the formation of an enzyme-substrate complex, immunogenic activity such as in an antigen-antibody reaction, as well as direct or indirect concentration of a particular analyte. It includes any chemical or physical interaction capable of liberating, forming or producing a detectable dye or the like which is indicative of its presence or concentration.

The dry analytical element may be of any type as long as it can be measured colorimetrically, regardless of detection component, type of reaction, constitution, measurement wavelength, etc. Not suitable for the invention. 2 dry analytical elements
The number of units (pieces) or more is preferable in order to improve analysis accuracy. When the reference liquid is spotted on another dry analytical element at the same time as the spotting of the liquid sample, the analytical accuracy can be further enhanced.

The dry analytical element comprises a liquid permeable liquid containing at least a portion of a reagent on a transparent, water impermeable support such as those described in US Pat. Nos. 3,992,158 and 4,292,272. It is preferably an integrated multi-layer analytical slide in which a layer and a porous layer are sequentially provided.

Dry analytical elements are also described in US Pat. No. 3,983,005.
No. 4,042,335, No. 4,066,403, No. 4,144, 306, No. 4,132,528,
4,258,001, 4,357,363, 4,381,42
It may have a structure as described in the No. 1 specification and the like. Further, it may have a plurality of porous layers as described in JP-A-61-4959, JP-A-62-138756, JP-A-62-138757, and JP-A-62-138758. .

When whole blood is used as the liquid sample, the dry analytical element preferably has a layer for filtering out substantially all or at least a part of blood cells. A liquid spreading layer may have this effect at the same time, see US Pat. No. 4,292,272.
Textile materials such as the woven fabrics described in the specification and the knitted fabrics described in JP-A-60-222769 are useful.

The liquid spreading layer is composed of any suitable fibrous or non-fibrous material, or mixture thereof, having a suitable porosity and average pore size capable of absorbing whole blood.
The liquid spreading layer preferably provides a uniform sample liquid per unit area on the surface facing the adjacent water-permeable layer with which it is in liquid contact. Useful liquid spreading layers include the fabrics described in U.S. Pat. No. 4,292,272 and JP-A-60-222.
It can be made of a fiber material such as a knitted fabric described in Japanese Patent Publication No. 769. It can also be constructed using a non-fiber isotropic porous material (for example, brush polymer) described in US Pat. No. 3,992,158.

In the liquid spreading layer, in order to control the spreading area, the spreading speed, etc., there are disclosed in JP-A-60-222770 and Japanese Patent Application No. 61-12.
It may contain a hydrophilic polymer or a surfactant as described in the specifications of No. 2875, No. 61-122876, No. 61-143754.

Dry analytical elements are described, for example, in US Pat. No. 4,042,33.
It may have a light shielding layer or a filtration layer as described in No. 5 specification. The dry analytical element may have a layer which, apart from the liquid spreading layer, substantially filters out whole blood cells. JP 58-70163 A, JP 61-4959 A, Japanese Patent Application 6
0-256408, 60-279859, 60-279
Porous layers such as those described in 860, 61-279861, etc. are suitable.

The dry analytical element may have a light reflecting layer. For example, a light reflecting layer can be provided between the reagent layer and the detection layer or between the reagent layer and the liquid spreading layer. The light-reflecting layer is a sample that is spot-supplied and supplied to the developing layer when photometrically measuring a detectable change (color change, color development, etc.) that has occurred in the detection layer, reagent layer, etc. from the side of the support having optical transparency. Liquid color,
In particular, when the sample is whole blood, it blocks the red color of hemoglobin, the yellow color of bilirubin, and the like, and also functions as a light reflection layer or a background layer. The light reflecting layer is preferably a water-permeable layer in which light reflecting fine particles such as titanium oxide and barium sulfate are dispersed using a hydrophilic polymer as a binder. As the binder, gelatin, gelatin derivative, polyacrylamide and the like are preferable. Hardeners may be added to curable polymers such as gelatin. If necessary, the dry analysis element may contain light-reflective particles such as titanium oxide in the developing layer, the reagent layer, the detecting layer and the like.

The dry analytical element may have a light-transmissive or transparent support. The support may be liquid permeable, such as paper, or liquid impermeable, such as plastic.

When a light transmissive support is used, the practically preferable constitution of the dry analytical element is as follows. (1) A substrate having a reagent layer on a support and a liquid spreading layer on the reagent layer. (2) A support having a detection layer, a reagent layer, and a liquid spreading layer in this order. (3) A support having a reagent layer, a light reflecting layer, and a liquid spreading layer in this order on the support. (4) A support having a detection layer, a reagent layer, a light reflection layer, and a liquid development layer in this order on a support. (5) A support having a detection layer, a light reflection layer, a reagent layer, and a liquid spreading layer in this order on the support.

In the above (1) or (3), a water absorbing layer may be provided between the support and the reagent layer. In the above (1) to (3), a blood cell filtration or other filtration layer may be provided between the reagent layer and the detection layer or the liquid development layer. (3) or above
In (5), between the light reflection layer and the detection layer, the reagent layer or the liquid development layer, between the reagent layer and the detection layer, or between the reagent layer and the liquid development layer, a blood cell filtration layer or other filtration layer. May be provided.

The one or more layers of the dry analytical element comprise one or more of various other optional ingredients such as surfactants, color former solvents, buffers, binders, hardeners. be able to. These components can be added in the same amount as that usually added. Typical element components are, for example, U.S. Pat.Nos. 3,992,158 and 4,042,335.
No.4, No.4,144,306, No.4,132,528, No.4,050,898, No.4,258,011 and No.4,
275,152 and 4,292,272.

The reagent composition contained in the dry analytical element is a composition capable of producing a substance that can be optically detected, for example, a dye, in the presence of a test component. Compositions that produce dyes by oxidation of leuco dyes (aryl imidazole leuco dyes as described in U.S. Pat. No. 4,089,747, JP-A-59-193352, etc.), diazonium salts, cups with other compounds when oxidized A composition containing a compound that produces a dye by ring (for example, 4-aminoantipyrines and phenols or naphthols), comprising a compound capable of producing a dye in the presence of a reduced coenzyme and an electron transfer agent Etc. can be used.

Examples of leuco dyes include US Pat. No. 4,089,
Examples thereof include triarylimidazole leuco dyes described in JP-A No. 747 and JP-A-59-193352, known triarylmethane leuco dyes, and the like.

The dye may be formed by a dye-forming composition containing a condensation product of an oxidizable compound and a color former. Examples of oxidizable compounds include benzidine and its homologues, p-phenylenediaminos, p-aminophenols, aminoantipyrines, such as 4-aminoantipyrine.

In the case of a dry analytical element for measuring enzyme activity, for example, a self-developing substrate (for example, para-nitrophenyl phosphate, para-nitrophenyl malto) which can release a colored substance such as p-nitrophenol. Pentaose, γ-glutamylparanitroanilide) can be contained in the reagent layer or the developing layer.

The reagent composition may contain an enzyme, and those described on pages 18 to 20 of the specification of JP-A-62-138756 can be used. Alternatively, an immunological reaction may be used (JP-A-64-88156, JP-A-1-112159, etc.). The reagent composition may be wholly or partly contained in a substantially uniform layer having a hydrophilic polymer as a binder. Examples of the hydrophilic polymer include gelatin and derivatives thereof (eg phthalated gelatin), cellulose derivatives (eg hydroxypropyl cellulose, carboxymethyl cellulose), agarose, acrylamide polymer, methacrylamide polymer, acrylamide or methacrylamide and various vinyls. A copolymer with a polymerizable monomer, polyvinylpyrrolidone, a copolymer of polyvinylpyrrolidone and various vinylic monomers, and the like can be used.

A part or all of the reagent composition may be contained in the porous layer. In order to allow at least one of the porous layers to contain a reagent composition that develops color in the presence of a component to be tested, a porous spreading layer pre-impregnated or coated with a suitable solution or dispersion of the reagent composition is used. The method of adhering to the water permeable layer, such as the reagent layer, by the method described in JP-A-55-164356 can be used.

The porous layer is adhered onto another water-permeable layer (eg, subbing layer, adhesive layer, water-absorbing layer) by the method described in JP-A-55-164356 and then the reagent composition. You may apply the solution or dispersion of above to a porous layer. A known method can be used for impregnating or coating the porous layer. For coating, for example, dip coating, doctor coating, hopper coating, and curtain coating are appropriately selected and used.

The reagent composition may be contained in one porous layer, for example, in the porous layer closest to the support among the two or more porous layers, or may be contained in two or more porous layers separately. You may let me.

The analytical element which does not contain a part or all of the reagents does not contain a part or all of the above-mentioned various reagents. Reagents that are commonly incorporated regardless of analysis items are preferably incorporated in advance. Such common reagents include pH buffers, color formers, stabilizers such as enzymes, and the like.

The target component (analyte) of the liquid sample to be detected may be a low molecular weight substance, an ion or a high molecular weight substance, and may be a hydrophilic substance or a hydrophobic substance. The component to be detected may be an enzyme, an antigen or an antibody. The antigen or antibody may be labeled with an enzyme or a fluorescent substance, or may be bound to another polymer.
The antigen may be a hapten.

[Examples of Analytes] Low molecular weight: urea, uric acid, ammonia, creatinine, lactic acid, pyruvic acid, glucose, galactose, neutral fat, cholesterol, hemoglobin, bilirubin ion: calcium, potassium, sodium, chloride ion polymer : Proteins (albumin, globulin, etc.), nucleic acids, lipoprotein enzymes: Amylase, alkaline phosphatase, lipase, lactate dehydrogenase, creatine phosphokinase, alanine aminotransferase, asparagine aminotransferase

Among the above-mentioned analytes, those which are preferably analytical elements in which all reagents are incorporated in advance are:
It is an item with high measurement frequency and high urgency. For example, ammonia, urea nitrogen, glucose, amylase,
Alanine aminotransferase and asparagine aminotransferase.

It is preferable to use an analytical element that does not include a part or all of the reagent because the measurement frequency is not so high and radical reaction conditions such as high pH and low pH are required, so that the reagent is built-in. This is an item that is difficult to make into a model slide. For example, total protein, alkaline phosphatase, ionized calcium and the like.

A typical example of analyzing a liquid sample using the analyzer of the present invention will be described. First, the analysis slides on which the components to be measured are displayed by identification marks such as bar codes are arranged in a predetermined analysis item order and loaded on a stacker or the like. . The analysis slides are taken out one by one in the stacking order and conveyed, and first, the identification mark is read by the identification mark reading device. Next, a predetermined amount of the liquid sample is spotted on the analysis slide, a predetermined reagent is selected according to the identification mark and a predetermined amount is spotted, or incubation is performed without the reagent spotted. The identification mark may be read before the spotting of the reagent or after the spotting of the liquid sample. on the other hand,
The liquid sample may be spotted after the reagent is spotted. After the incubation, the optical density of the color reaction portion of the analytical element is reflected and measured by a conventional method, and the density is calculated from the calibration curve obtained in advance.

[0039]

[Example] As the analyzer, a commercially available dry analysis slide analyzer (“Fuji Drychem 5000”, a product of Fuji Photo Film Co., Ltd.) was used, and a fixed amount supply device 1 for sample liquid and a fixed amount of reagent liquid as shown in FIG. The supply device 2 was attached and used. Both of the constant quantity supply devices 1 and 2 are substantially the same type, and both are composed of turntables 3 and 4 and dispensing arms 5 and 6. On the peripheral portion of the turntable, cups 7 for containing a sample liquid or a reagent liquid and microchips 8 for quantitative supply are alternately arranged. By rotating and lowering the dispensing arms 5 and 6, the microchip 8 arranged on the turntables 3 and 4 is fitted to the mounting rod 9 projecting downward from the rotating end, and after rising, The turntables 3 and 4 are slightly rotated so that the cup 7 adjacent to the chip 8 is positioned directly below the cup 7. Then, the dispensing arms 5 and 6 descend,
A certain amount of the sample liquid or the reagent liquid contained in the cup 7 is contained in the microchip 8 and then raised again. Next, the dispensing arm pivots largely and moves to a position right above the analysis slide 10, where it descends and spots the sample liquid or reagent liquid, rises again, and returns to the original position to detach the microchip. . The above operation is repeated.

An analytical element was prepared in which a hydrophilic polymer layer and a porous spreading layer were laminated in this order on a transparent support, and an analytical element containing no reagent was prepared and narrowed into a frame to give an analytical slide. On the other hand, an alkaline phosphatase measurement reagent solution having the following composition was prepared. 2-Amino-2-methyl-1-propanol 89.1mg Disodium paranitrophenyl phosphate 370mg Magnesium chloride 4mg Water 10g (pH 9.9)

Controlled Serum CPL, M, H for Fuji Dry Chem
A calibration curve for alkaline phosphatase was obtained by using.

Acetylcholinesterase, total protein and glutamate pyruvate transaminase (GPT) were measured using the above analyzer. The above analysis slides were used for acetylcholinesterase analysis, and commercially available analysis slides containing all reagents (“Fuji Dry Chem Slide T”) were used for analysis of total protein and GPT.
P ”and“ Fuji Dry Chem Slide GPT ”, both of which are products of Fuji Photo Film Co., Ltd. Ten slides of each of the above three types were randomly stacked on the stacker in the analyzer and the measurement was started. Each slide taken out from the stacker was moved to the sample spotting station of the analyzer and stopped there. When the analysis slide is for acetylcholinesterase, first, the reagent liquid quantitative supply device 2 is activated to deposit 10 μl of the reagent liquid, and after 5 seconds, the sample liquid quantitative supply device 1 is activated to operate the vein surface of a healthy person. Heparin was added to and the serum obtained by centrifugation was spotted at 10 μl. Analysis slide for total protein or G
In the case of PT, the constant amount supply device 2 of the reagent solution was not operated, and only the above-mentioned serum as the sample solution was spotted. Each slide was incubated with the above analyzer at 37 ° C. for 6 minutes, the reflection optical density at 400 nm was measured, the density was determined by a calibration curve, and the reproducibility of the measured value was examined. The results are shown below. Average S D C V Alkaline phosphatase 22u / l 4.8u / l 2.2% Total protein 7.4g / dl 0.9 1.2% GPT 18u / l 3.9 2.2%

[0043]

According to the present invention, the routine inspection and the special inspection can be performed by one analyzer, and the analysis request peculiar to the facility can be met. The analytical element that does not include some or all of the reagents can be used in the same manner as conventional analytical elements by allowing the user to spot and dry the reagents. Therefore, it can be used as a user's own analysis method. In addition, the reagent can be spotted in two or more parts, whereby the endogenous interfering substances such as ascorbic acid, pyruvic acid, bilirubin, etc. are removed before the detection reaction is performed, or during storage. Acids, alkalis, heavy metals, etc. that cause deterioration of the enzyme reagent can be stored separately.

[Brief description of drawings]

FIG. 1 is a diagram illustrating a usage state of a main part of an analyzer that is an embodiment of the present invention.

Claims (2)

[Claims] 1. An analyzer for a dry analytical element, comprising at least a fixed amount supply device for a sample liquid and a fixed amount supply device for a reagent liquid. 2. An analytical method for measuring at least one type of analytical element in which all reagents are incorporated and at least one type of analytical element lacking a part or all of the reagents, and these analytical elements are provided. Measuring method, characterized in that a reagent that is lacking when measuring