WO2013122121A1 - Test instrument for immunochromatography, analyzer and analysis method - Google Patents

Test instrument for immunochromatography, analyzer and analysis method Download PDF

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Publication number
WO2013122121A1
WO2013122121A1 PCT/JP2013/053463 JP2013053463W WO2013122121A1 WO 2013122121 A1 WO2013122121 A1 WO 2013122121A1 JP 2013053463 W JP2013053463 W JP 2013053463W WO 2013122121 A1 WO2013122121 A1 WO 2013122121A1
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substance
detection unit
test instrument
signal value
immunochromatographic test
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PCT/JP2013/053463
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French (fr)
Japanese (ja)
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裕一郎 清水
中野　郁雄
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シャープ株式会社
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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
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/558—Immunoassay; Biospecific binding assay; Materials therefor using diffusion or migration of antigen or antibody
    • 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
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
    • G01N33/54366—Apparatus specially adapted for solid-phase testing
    • G01N33/54386—Analytical elements
    • G01N33/54387—Immunochromatographic test strips
    • G01N33/54388—Immunochromatographic test strips based on lateral flow

Definitions

  • the present invention relates to an immunochromatographic test instrument, an analysis apparatus, and an analysis method.
  • the present invention detects an immunochromatographic test instrument (chip) for detecting and analyzing a specific component (for example, antigen, antibody, enzyme, substrate, cytokine, etc.) contained in blood, an analyzer, and the above component And how to analyze.
  • a specific component for example, antigen, antibody, enzyme, substrate, cytokine, etc.
  • Immunochromatography is a measurement method for detecting a sample containing a substance to be detected simply and in a short time by an antigen-antibody reaction. Therefore, it is now widely used in many scenes, such as clinical examinations in hospitals and certification tests in laboratories. Utilizing these characteristics, immunochromatography is used for pregnancy test drugs and influenza test drugs, and is attracting attention as a new POCT (Point Of Care Testing) method.
  • POCT Point Of Care Testing
  • POCT refers to a test for diagnosis as close as possible to the patient. Therefore, according to POCT, prompt and accurate treatment is possible based on examination information provided instantaneously. Therefore, an emergency inspection at a hospital and an inspection during an operation become possible, and the needs in the medical field are increasing.
  • Fig. 4 shows the structure of an immunochromatographic test instrument in a general immunochromatography method.
  • the conjugation pad 108 is arranged so that the first capture substance 110 (antibody-sensitized latex 109) to which the substance 105 to be detected is bound and the label substance is bound can be moved.
  • the second capture substance 104 that captures the substance 105 to be detected includes a detection unit 103 fixed on the porous body 101, an absorption pad 107 that absorbs liquid, and a sample pad 102 that drops a specimen.
  • Patent Document 1 discloses providing a plurality of detection units. Using this, it is disclosed to perform semi-quantification from the number of colored detection parts after reaction.
  • Patent Document 2 a site for immobilizing an antibody that specifically binds to a labeled antibody that is irrelevant to a substance to be detected is provided, and a labeled antibody prepared so that a constant signal value is obtained every time measurement is performed. It is disclosed. Using this, it is disclosed that quantification is performed by comparing the signal value of the site where the antibody that specifically binds to the labeled antibody unrelated to the substance to be detected after the reaction is immobilized with the signal value of the detection unit. ing.
  • Patent Document 3 discloses providing a standard color region in which particles are directly fixed in advance. Using this, it is disclosed that quantification is performed by comparing a standard color region after reaction with a detection unit.
  • Patent Document 4 discloses that the length of the detection unit is increased. Using this, it is disclosed that it quantifies by reading the length of the colored region after reaction.
  • Patent Document 5 discloses a system provided with a temperature control unit. Since the reactivity changes depending on the temperature, it is disclosed that the quantitativeness is improved by keeping the temperature constant.
  • Japanese Patent Publication “Japanese Patent Laid-Open No. 05-005743” (published on January 14, 1993) Japanese Patent Publication “Japanese Patent Laid-Open No. 2001-83153 (published on March 30, 2001)” Japanese Patent Publication “International Publication No. 2006/080438 (Released on August 3, 2006)” Japanese Patent Publication “Japanese Patent Laid-Open No. 2010-025887 (published on February 4, 2010)” Japanese Patent Publication “JP 2011-128038 A” (published on June 30, 2011)
  • the density and thickness of the porous body itself are not strictly constant at the site to be used, so that the amount of the second capture substance is not constant between instruments. . Specifically, since the amount and density of the second trapping substance itself vary between instruments, the degree of coloration is not strictly constant.
  • Patent Documents 2 and 3 although not specified, there is a possibility that the density and thickness of the porous body itself can be considered.
  • the devices described in Patent Documents 2 and 3 are not easy to manufacture and are complicated.
  • an antibody that specifically binds to a labeled antibody unrelated to the specimen is used as described in Patent Document 2, it is a substance that does not inhibit the reactivity of the antibody for detecting the specimen, and the background.
  • the material must not be an element.
  • the problem of the material is the same, but in addition, a device for keeping the coloring degree constant is required.
  • the present invention has been made in view of the above-described problems, and an object of the present invention is to provide an immunochromatographic test instrument and analyzer capable of simply measuring a stable quantitative value using an instrument using an immunochromatography method. And providing an analysis method.
  • the present invention provides a first capture in which a substance to be detected is captured and a labeling substance is bound in an immunochromatographic test instrument having a porous body in which a specimen can move by capillary action. An area where the substance is movable, a detection part where the second capture substance that specifically captures the substance to be detected is fixed on the porous body, and the porous And a reference detection unit in which a third capture substance for capturing the first capture substance is disposed, and the third capture substance is saturated.
  • the first capturing substance is captured by adsorption.
  • the third capture substance captures the first capture substance with respect to the porous body so as to capture the maximum amount of the first capture substance that can be adsorbed by the third capture substance. Material is being washed away.
  • saturated adsorption means a state where the third capture substance does not adsorb to the first capture substance even if the first capture substance is further introduced.
  • the signal value of the detection unit varies depending on the thickness and density of the porous body, as well as the value varies depending on the concentration of the target substance (substance to be detected) contained in the specimen. In addition, the signal value varies depending on the storage environment and use environment. With the above configuration, since the signal value of the reference detection unit always detects a signal value corresponding to the amount of the first capture substance captured by saturation adsorption (always a constant amount), the signal of the reference detection unit It is possible to eliminate the influence of the above factors by referring to the values.
  • the signal value changes due to different local thickness and density.
  • the signal for example, fluorescence
  • the signal which a labeling substance emits changes with various conditions. For example, if the labeling substance becomes old, the signal emitted by the labeling substance becomes weaker with time. For these reasons, the signal emitted by a certain amount of labeling substance is not always constant.
  • the third capture substance can always bind a certain amount of the first capture substance, even if the labeling substance is old, it is constant at the time of detection.
  • a signal eg, fluorescence intensity
  • the amount can be calculated accurately. Since the first capture substance captures the substance to be detected, if the amount of the first capture substance captured by the detection unit can be accurately calculated, the detection should be detected by the detection unit. The amount of substance can also be calculated accurately.
  • the reference detection unit is provided on the downstream side of the detection unit.
  • downstream is downstream of the flow in which the specimen moves by capillary action.
  • the amount of the first capture substance flowing through the porous body and the amount of the labeling substance bound to the first capture substance are both the second capture substance. It is preferable that the amount is equal to or greater than the sum of the amount of the third trapping substance and the amount of the third trapping substance.
  • the amount of the first trapping substance can be easily adjusted because the amount of the first trapping substance is satisfied so that the reference detection unit always captures saturated adsorption. it can. Moreover, according to the said structure, a target substance can be measured more correctly.
  • the porous body is branched, and the detection section is provided in one of the branched porous bodies.
  • the detection section is provided. It is preferable that the reference detection unit is provided in a porous body different from the porous body provided with.
  • the amount of the first capture substance flowing into the porous body and the amount of the labeling substance bound to the first capture substance are respectively determined by the second detection unit. It is preferable that the amount is not less than twice the amount of the capture substance or the amount of the third capture substance in the reference detection unit. In particular, the amount of the first capture substance flowing into the porous body and the amount of the labeling substance bound to the first capture substance are the amount of the second capture substance in the detection unit and the reference, respectively. It is preferable that the amount is more than twice the relatively large amount of the third capturing substance in the detection unit.
  • the amount of the first capture substance can be easily adjusted because the amount of the first capture substance is satisfied so that the reference detection unit always captures the saturated adsorption. . Moreover, according to the said structure, a target substance can be measured more correctly.
  • the reference detection unit and the detection unit preferably include the same detection means.
  • the detection is performed by the same detection means, even if the detector itself is affected by a change in the usage environment such as temperature, the fluctuation range of the signal value in the detection unit and the reference detection unit is the same. Therefore, the influence can be eliminated.
  • the reference detection unit and the detection unit preferably have a structure capable of observing color shading or light emission from the outside.
  • the target substance can be optically detected based on color shading or light emission.
  • the target substance to be detected optically may be one that develops color or emits light, or may be one that is modified with a substance that produces or emits light.
  • the shading of the color due to the coloring substance may be optically detected.
  • light emission by the light emitting substance may be optically detected.
  • the light emission includes chemical reaction light emission and fluorescence (light emission by photoexcitation).
  • the reference detection unit and the detection unit preferably include a working electrode and a reference electrode, respectively.
  • the target substance can be detected electrochemically in the detection unit.
  • the target substance to be detected electrochemically may be one that is electrochemically active per se, or one that is modified with an electrochemically active substance.
  • a current value obtained from an electrochemically active substance may be measured with a detection electrode.
  • the working electrode and the reference electrode have the same size.
  • the first capture substance is preferably an antibody against the substance to be detected.
  • a substance to be detected by an analyzer of an immunochromatographic test instrument used for biochemical analysis is an in vivo protein.
  • Antibodies that are difficult to denature are optimal substances for use as capture substances.
  • the second capture substance is preferably an antibody against the substance to be detected.
  • a substance to be detected by an analyzer of an immunochromatographic test instrument used for biochemical analysis is an in vivo protein.
  • Antibodies that are difficult to denature are optimal substances for use as capture substances.
  • the third capture substance is preferably an antibody against the first capture substance.
  • a substance to be detected by an analyzer of an immunochromatographic test instrument used for biochemical analysis is an in vivo protein.
  • Antibodies that are difficult to denature are optimal substances for use as capture substances.
  • the labeling substance is preferably a fine particle.
  • the amount of the first capture substance can be detected as the amount of the fine particles.
  • the amount of fine particles can be detected by, for example, color shading.
  • the labeling substance is preferably a fluorescent material.
  • the labeling substance is preferably an electrochemically active substance.
  • the electrochemically active substance is used as the labeling material, it can be easily detected by direct electrochemical detection.
  • a determination unit in which a fourth capture substance for capturing the first capture substance is disposed downstream of the reference detection unit.
  • the immunochromatographic test instrument In the immunochromatographic test instrument according to the present invention, information on the correlation between the detection signal value derived from the signal value of the detection unit and the signal value of the reference detection unit and the concentration of the substance to be detected is written.
  • a bar code or IC tag is preferably provided.
  • information can be written in advance on the chip.
  • the information to be written can be changed according to the lot at the time of manufacture and the lot of the reagent.
  • the analyzer according to the present invention is characterized by including a detector that detects a signal emitted from the labeling substance in the immunochromatographic test instrument according to the present invention.
  • the analyzer it is preferable to calculate the concentration of the substance to be detected from a relative signal value obtained by comparing the signal value of the detection unit and the signal value of the reference detection unit.
  • the calculation unit calculates the concentration of the substance to be detected from the relative signal value obtained by comparing the signal value of the detection unit and the signal value of the reference detection unit.
  • the signal value depending on the concentration of the target substance can be obtained by offsetting the influence of the elapsed time, storage state, and usage environment after chip manufacture, for example, by taking the ratio with the signal value of the reference detection unit. Can be obtained.
  • any correction means such as a difference or a correction coefficient is applied.
  • the analyzer according to the present invention preferably issues a warning when the signal value of the reference detection unit is equal to or less than a reference value.
  • the warning generation unit issues a warning when the signal value of the reference detection unit is equal to or less than a reference value.
  • the detector to detect the signal value of the detection unit and the signal value of the reference detection unit is the same.
  • a common detector can be used for the signal value of the detection unit and the signal value of the reference detection unit, thereby simplifying the configuration of the analyzer. be able to.
  • the analyzer according to the present invention is preferably provided with a reader for barcode or IC tag.
  • the said structure it will become possible to read information from the barcode provided in the inspection instrument for immunochromatography or an IC tag.
  • the read information can be used for calibration.
  • the analysis method according to the present invention uses the immunochromatographic test instrument of the present invention, compares the signal value of the reference detection unit and the signal value of the detection unit, and determines the detection signal value. It is characterized by calculating.
  • concentration of the said substance which should be detected from the relative signal value which compared the signal value of a reference detection part with the signal value of a reference detection part and the signal value of a detection part about the time passage after a chip manufacture, a storage state, and a use environment. It is possible to calculate.
  • the detection signal value is calculated from a ratio between the signal value of the detection unit and the signal value of the reference detection unit.
  • the correlation between the detection signal value and the concentration of the substance to be detected is previously input to the immunochromatographic test instrument, and the detection should be performed from the detection signal value and the correlation. It is preferable to calculate the concentration of the substance.
  • the detection signal value is calculated using the reference detection unit and the detection unit, a value close to the actual value can be output.
  • the correlation based on information relating to at least one of a production lot and a reagent lot of the immunochromatographic test instrument.
  • the information is read from a barcode or an IC tag provided on the immunochromatographic test instrument.
  • the present invention quantitative detection is possible with an immunochromatographic test instrument without depending on the non-uniformity of the porous body itself used.
  • the original accurate value can be detected without depending on the storage state, the period until use or the use environment.
  • the original accurate value can be detected by an element or device having a simple configuration.
  • sample refers to a specimen (test object) introduced into an immunochromatographic test instrument, whether or not it includes a target substance (substance to be detected) to be detected. May be.
  • the term “capture substance” refers to a substance that forms a covalent bond or a non-covalent bond with the target substance by specifically interacting with the target substance.
  • the capture substance is specifically a substance having a relationship between a host and a guest with the target substance.
  • Examples of the capture substance include antigens, antibodies, enzymes, substrates, ligands, receptors, DNA, sugars, peptides And synthetic polymers (for example, molecular imprint polymer).
  • upstream and downstream are concepts based on the flow of fluid in the porous body, and the direction in which the liquid flows is referred to as “downstream”, and the direction opposite to the flow of the liquid is referred to as upstream. .
  • upstream the sample pad side in the porous body is “upstream”.
  • FIG. 1 is a schematic diagram of an immunochromatographic test instrument according to Embodiment 1 of the present invention.
  • the lower end of the absorbent pad 7 is placed on the backing sheet 15 whose surface is coated with an adhesive, the upper end of the porous body 1, the upper end of the conjugation pad 8 and the porous body 1.
  • the upper end of the sample pad 2 are pasted so as to overlap with the lower end of the conjugation pad 8 by several millimeters. After pasting, it can be cut into a strip shape by cutting with a width of several tens of ⁇ m to several cm. A width of several hundred ⁇ m to several mm is preferable from the viewpoint of the amount of specimen.
  • each cut instrument can be placed in the housing to make it easy for the user to use.
  • the backing sheet 15 has a function as a substrate for combining the materials.
  • the absorption pad 7 has a function of absorbing the specimen flowing through the porous body 1 by capillary force.
  • the conjugation pad 8 is an area where a first capture substance 9 (for example, antibody-sensitized latex particles) to which a label substance is bound is stored, and is stored so that it can move when a liquid passes through.
  • the sample pad 2 indicates an area for injecting a specimen.
  • the detection part 3 is provided on the porous body.
  • a second capture substance 4 that captures a substance to be detected and analyzed is immobilized on the detection unit 3.
  • a labeled first capture substance 9 stored in the conjugation pad 8 and a reference detection unit 5 for evaluating the activity of the label substance are provided in the porous body 1.
  • a third capture substance 6 that captures the first capture substance is immobilized on the reference detection unit 5.
  • the detection unit 3 and the reference detection unit 5 may be provided anywhere within the porous body 1, but the detection unit 3 may be provided on the sample pad 2 side and the reference detection unit 5 may be provided on the absorption pad 7 side. It is preferable from the viewpoint of reactivity.
  • the material of the porous body 1 is not limited as long as the medium can move the liquid by capillary action.
  • woven fabrics, nonwoven fabrics, and the like made of porous synthetic polymers such as cellulose, cellulose derivatives, nitrocellulose, ethylene vinyl acetate, polyurethane, polymethyl methacrylate, nylon resin, polyvinylidene difluoride (PVDF), etc.
  • porous synthetic polymers such as cellulose, cellulose derivatives, nitrocellulose, ethylene vinyl acetate, polyurethane, polymethyl methacrylate, nylon resin, polyvinylidene difluoride (PVDF), etc.
  • PVDF polyvinylidene difluoride
  • the sample pad 2 is a part where the specimen is dropped. Further, when an insoluble substance is present in the specimen, it also functions as a removal filter.
  • the material that can be used for the sample pad 2 is not particularly limited as long as it is hydrophilic and has a filter function. For example, regenerated cellulose, cellulose acetate, nitrocellulose, polyacrylonitrile, ethylene vinyl acetate, polyurethane, poly Examples include methyl methacrylate, nylon resin, glass fiber, pulp, cotton, rayon, acrylic, and polyester.
  • the detection unit 3 is a part that detects a substance in the fluid flowing through the porous body 1.
  • the detection unit 3 includes a substance (hereinafter, referred to as a target of detection and analysis).
  • a second capture substance 4 that captures the target substance is immobilized.
  • the second capture substance 4 is a substance (for example, an antibody, an antigen, a peptide, DNA, a sugar, a synthetic polymer (for example, a molecular imprint polymer), an enzyme, a substrate, a ligand, or a receptor that has a host-guest relationship with the target substance.
  • an antibody or a synthetic polymer is preferable because its activity is stable.
  • a known method such as a physical adsorption method, a chemical bonding method, or a covalent bonding method can be appropriately employed.
  • a spotter for immobilization In particular, by performing a line plot, it is possible to quantitatively drop the antibody solution in a line shape. After immobilization, the porous body 1 is dried by air drying or hot air drying.
  • the configuration of the detection unit 3 is not particularly limited, and can be appropriately determined depending on the detection method of the target substance.
  • the structure may be such that the color can be observed from the outside.
  • detection is possible by providing an observation window.
  • the detection unit 3 only needs to include detection means having a detection electrode formed on the top or bottom of the porous body 1.
  • the detection electrode only needs to be composed of at least two electrodes, that is, a reference electrode and a working electrode, but is preferably composed of three electrodes provided with a counter electrode in addition to the reference electrode and the working electrode. If there are three electrodes, the reference potential can be taken, so that more accurate measurement can be performed.
  • the reference electrode, the working electrode, and the counter electrode can be formed by a microfabrication technique using a conventional photolithography technique.
  • the conductive material of the electrode for example, gold, platinum, silver, chromium, titanium, iridium, copper, or carbon can be used.
  • a silver / silver chloride electrode is preferably used from the viewpoint of stability of the standard potential.
  • the reference electrode, the working electrode, and the counter electrode have the same size. According to the above configuration, the signal values can be compared without considering the size of the diffusion layer spreading on the electrode interface.
  • the reference detection unit 5 is a site for evaluating the activity of the labeled first capture substance 9 and the label substance stored in the chip. As shown in FIG. 1, a third capture substance 6 for capturing the first capture substance is immobilized on the reference detection section 5 provided between the detection section 3 and the absorption pad 7. .
  • the third capture substance 6 is a substance having a host-guest relationship with the first capture substance (eg, antigen, antibody, enzyme, substrate, ligand, receptor, DNA, sugar , A peptide or a synthetic polymer (for example, a molecular imprint polymer), and the like.
  • the first capture substance eg, antigen, antibody, enzyme, substrate, ligand, receptor, DNA, sugar , A peptide or a synthetic polymer (for example, a molecular imprint polymer), and the like.
  • an antibody or a synthetic polymer is preferable because its activity is stable.
  • a known method such as a physical adsorption method, a chemical bonding method, or a covalent bonding method can be appropriately employed.
  • the porous body 1 is dried by air drying or hot air drying. After immobilizing the second capture substance and the third capture substance on the porous body 1, nonspecific adsorption can be suppressed by immersing the entire porous body 1 in the nonspecific adsorption inhibitor.
  • the nonspecific adsorption inhibitor for example, commercially available nonspecific adsorption inhibitor such as bovine serum albumin (BSA), casein, gelatin, an electrically neutral polymer or a polymer having a phosphorylcholine group can be used. After the blocking treatment, sufficient drying is performed by natural drying, air drying, hot air drying or reduced pressure drying.
  • the configuration of the reference detection unit 5 is the same as that of the detection unit 3 and is not particularly limited, and can be determined as appropriate depending on the detection method of the target substance.
  • the structure may be such that the color can be observed from the outside.
  • detection is possible by providing an observation window.
  • the reference detection unit 5 only needs to include detection means having detection electrodes formed on the upper or lower portion of the porous body 1.
  • the detection electrode only needs to be composed of at least two electrodes, that is, a reference electrode and a working electrode, but is preferably composed of three electrodes provided with a counter electrode in addition to the reference electrode and the working electrode.
  • the configuration of the reference detection unit 5 uses the same detection means and / or detection structure as the detection unit 3. If the detection means and / or the detection structure are the same, the influence of detection variation can be ignored.
  • the detection signal value also varies depending on the temperature. However, if the detection means and / or the detection structure are the same, the fluctuation range can be regarded as the same.
  • the labeling substance of the labeled first capturing substance 10 for detecting the substance in the fluid is used as a labeling agent such as latex particles, colloidal metals, dyes, fluorescent dyes (or fluorescent proteins), enzymes or radioactive substances. Any substance can be used.
  • the labeling substance may be fine particles. According to the above configuration, the amount of fine particles can be detected based on color shading.
  • the particle size of the “fine particles” in the present specification is not particularly limited as long as it is 50 ⁇ m or less, but is preferably 0.01 to 2 ⁇ m.
  • the labeling substance may be an electrochemically active substance. According to the said structure, it can detect electrochemically with a detection electrode. Examples of the electrochemically active substance include ferricyanide, ferrocyanide, ferrocene and ferrocene derivatives.
  • a substance having a host-guest relationship as described above for example, antigen, antibody, enzyme, substrate, ligand, receptor, DNA, sugar, peptide, or synthetic polymer (for example, molecular imprint polymer)
  • an antibody or a synthetic polymer is preferable because its activity is stable.
  • the labeled first capture substance is temporarily stored by the conjugation pad 8.
  • the first capture substance temporarily stored by the specimen that has moved from the sample pad 2 due to capillary action is dissolved, and becomes a site for capturing the substance to be detected contained in the specimen.
  • the storage amount of the labeled first capture substance 10 is an amount that allows the first capture substance to react sufficiently in the detection unit 3 and an amount that can be reacted in a substantially saturated manner in the reference detection unit 5.
  • the amount of the first capture substance can be expressed by the following equation. .
  • the first capture substance can be prepared as follows. Since the amount of immobilization of the second capture material 4 and the third capture material 6 varies depending on the thickness or density of the porous body 1 itself, it is basically preferable to determine the conditions by this method. Specific preparation examples are shown below.
  • the sum or more of the amount of the first capture substance obtained from (1) and (2) is stored in the conjugation pad 8.
  • the recognition site of the third capture substance 6 recognizes a site different from the recognition site of the first capture substance 10
  • the first capture substance 10 associated with the substance to be detected is the third capture substance 10.
  • the trapping substance 6 There is a possibility of being trapped by the trapping substance 6.
  • the effects of the present application are not impaired.
  • the detection unit 3 detects the amount of the first capture substance 10 captured on the detection unit 3 (corresponding to the amount of the captured object), and the reference detection unit 5 detects the first captured substance 10 captured on the reference detection unit 5. The amount of one capture substance 10 is detected.
  • the detection signal value is expressed as follows.
  • the detection signal value is expressed as follows.
  • Detection signal ⁇ (formula 1) x enzyme substrate reaction rate (formula 2)
  • enzyme substrate reaction rate (formula 2)
  • the signal value of the detection unit 3 varies depending on the concentration of the target substance contained in the sample, but the signal value of the reference detection unit 5 is always detected by a signal value corresponding to saturated adsorption. Therefore, only the influence of the above factors can be considered.
  • the signal value of the reference detection unit 5 can be expressed as follows.
  • Equation 3 Signal value of the reference detection part ⁇ Immobilization density of the capture substance (third capture substance) (depending on the density of the porous body 1) ⁇ Thickness of the porous body 1 ⁇ antigen-antibody reaction rate (Equation 3) Compared with Equation 1 above, factors that are affected by the use environment can be canceled out, so the difference between the actual value can be reduced by comparing the signal value of the detection unit 3 and the signal value of the reference detection unit 5. Is possible.
  • the correction method can be performed by taking a ratio, but any other correction means such as applying a difference or a correction coefficient may be used.
  • the signal value of the detection unit 3 may be reduced by a maximum of 20%, but the signal value of the reference detection unit 5 also decreases, When compared, the influence of the porous body 1 itself can be ignored.
  • the signal value may change depending on the lot of the instrument. Therefore, it becomes possible for the user to respond by inputting correction information in advance to an apparatus or software for detecting an immunochromatographic test instrument in accordance with the lot change.
  • the reference detection unit 5 can also be used to determine whether or not the detection by the immunochromatographic test instrument has failed.
  • the reference detection unit 5 detects a signal value when the labeled first capture substance is saturated and adsorbed, but when the signal value is detected below a predetermined value, the reliability of the measurement itself is low. It is possible to judge.
  • the predetermined value can be used as a reference value.
  • a determination unit 20 in which a fourth trapping substance 21 (not shown) for trapping the first trapping substance is immobilized on the porous body 1 may be provided.
  • the determination unit 20 is provided on the downstream side of the reference detection unit 5.
  • the determination unit 20 it can be determined whether or not the reaction itself has been completed.
  • the amount of the first capture substance can be easily determined. Specifically, since the determination unit 20 needs to have a reaction that is detected as a detection signal by the labeling substance, the detection signal of the determination unit 20 can be obtained without taking the preparation method described in 1.5.2.
  • the storage amount of the first capture substance may be adjusted by an amount such that excess first capture substance that has not been captured by the detection unit 3 and the reference detection unit 5 flows to the determination unit 20. .
  • the fourth capture substance includes a substance having a host-guest relationship with the first capture substance (eg, antigen, antibody, enzyme, substrate, ligand, receptor, DNA, sugar, peptide, or synthetic polymer (eg, molecular in Print polymer) and the like.
  • the first capture substance eg, antigen, antibody, enzyme, substrate, ligand, receptor, DNA, sugar, peptide, or synthetic polymer (eg, molecular in Print polymer) and the like.
  • an antibody or a synthetic polymer is preferable because its activity is stable.
  • the fourth capture substance is preferably the same capture substance as the third capture substance.
  • a determination part can be formed with the structure similar to a reference detection part, and the structure of the test
  • Embodiment 2 A second embodiment of the present invention will be described with reference to FIG. In addition, about the same structure as Embodiment 1, the detailed description is abbreviate
  • the reference detection unit 5 and the detection unit 3 are arranged in series in the same porous body 1, but in the second embodiment, as shown in FIG. 3, the reference detection unit 5 ′ and the detection unit 3 are detected.
  • the part 3 ′ is arranged in parallel with the branched porous body 1.
  • the number of branches of the porous body 1 is not particularly limited and can be set as appropriate.
  • the number may be two, three, four, or more.
  • the number can be an even number (for example, 4, 6, 8,).
  • each detection unit 3 ′ and reference detection unit 5 ′ can be configured according to the substance to be detected and analyzed.
  • the plurality of detection units 3 ′ may be detection units 3 ′ having the same configuration, but may be detection units 3 ′ having different configurations.
  • the plurality of reference detection units 5 ′ may be reference detection units 5 ′ having the same configuration, but may be reference detection units 5 ′ having different configurations.
  • the size and shape of the cross section of each branched porous body in the direction perpendicular to the direction in which the fluid moves are not particularly limited, but are preferably the same size and shape. According to the above configuration, it is possible to detect and analyze a substance with high accuracy.
  • the storage amount of the labeled first capture substance is an amount that allows the first capture substance to react sufficiently in the detection unit 3 ′, and is almost saturated in the reference detection unit 5 ′. The amount can be reacted.
  • the immobilized amounts of the second capture substance 4 ′ (not shown) in the detection unit 3 ′ and the third capture substance 6 ′ (not shown) in the reference detection unit 5 ′ are obtained as data.
  • the amount of the first capture substance can be represented by the following formula:
  • the first capture substance when a capture substance that captures a large number of epitopes for one target substance, such as a polyclonal antibody, is used as the first capture substance, it is desirable to store the first capture substance about three times the above formula.
  • the first capture substance can be prepared as follows. It is. Specific preparation examples are shown below. (1) Confirmation of reference detection unit 5 ′ Using the first capture substance (standard substance) having a known concentration, the detection range of the first capture substance in the reference detection unit 5 ′ is examined. By deriving the product of the capacity from the obtained saturated concentration, the amount necessary for the first capture substance to be saturated and adsorbed in the reference detection unit 5 ′ can be calculated. (2) Confirmation of detection part 3 'The maximum amount of the first capture substance required in the detection part 3' is examined using a target substance (standard substance) whose concentration is known.
  • the concentration of the first capture substance necessary for saturated adsorption to the target substance captured on the detection unit 3 ′ is examined. .
  • the amount necessary for the first trapping substance to be saturated and adsorbed can be calculated.
  • the analyzer according to the present invention may have any configuration as long as it can realize the analysis method using the immunochromatographic test instrument according to the present invention, but the labeling substance in the immunochromatographic test instrument emits.
  • a detector detection means for detecting a signal and a calculation unit are provided.
  • the analyzer according to the present invention may be provided with the immunochromatographic test instrument according to the present invention.
  • the analyzer according to the present invention includes a detector that detects the signal value of the detection unit 3 and the signal value of the reference detection unit 5 of the immunochromatographic test instrument according to the present invention.
  • the configuration of the detector is not particularly limited, and can be appropriately determined according to the detection method of the target substance, similarly to the detection unit 3 and the reference detection unit 5.
  • the detection means in the detector corresponding to the detector 3 and the detector corresponding to the reference detector 5 are the same. If it is the said structure, since the same detection means is used, a common detector can be used and the structure of an analyzer can be simplified.
  • the analysis apparatus may include a detector that detects a signal from the determination unit. While no signal is detected from the determination unit, it is determined that the reaction has not ended, and the detection process is continued. If a predetermined signal is detected from the determination unit, it is determined that the reaction is completed, the detection process is terminated, and the calculation unit corrects the signal value and calculates the calibration value.
  • the calculation unit compares the signal value detected from the detection unit with the signal value detected from the reference detection unit according to the method described in ⁇ 1.7 Correction of Signal Value>, and detects the detection signal value ( Relative signal value) is calculated. Thereafter, a calibration value (concentration of a substance to be detected) can be calculated from a detection signal value obtained as a detection result based on a calibration curve, data information, or the like previously input to the apparatus side.
  • the analyzer according to the present invention may have the following configuration in addition to the above configuration.
  • the analyzer according to the present invention may include a display unit that displays a calibration value.
  • the display unit may be a display, or may print out a calculation result.
  • the analyzer according to the present invention preferably includes a warning generator.
  • a warning generator As described in ⁇ 1.8 Detection error>, when the signal value of the reference detection unit 5 falls below the standard value during detection by the analyzer, a warning is given to the user and the calibration value is displayed on the display unit. By avoiding this, it is possible to prevent an erroneous calibration value from being transmitted to the user.
  • the warning may be a light warning or a sound warning, and a conventionally known method can be used. However, when detection by light is performed in the detection unit and the reference detection unit, a warning by sound is preferable so as not to affect them.
  • the analyzer according to the present invention preferably includes a barcode or IC tag reader. Embedded in the IC tag, barcode, etc. of the chip are information for calibration such as differences in lots at the time of manufacture and differences in lots of reagents. On the analyzer side, it is possible to correct the calibration curve or the data by reflecting the information input to the barcode or IC tag on the calculation unit. Any known technique can be used as a method of reading a barcode or IC tag by the analyzer.
  • the present invention relates to an instrument, an apparatus and a method for quantitatively detecting a substance to be measured in a specimen using immunological chromatography (immunochromatography).

Abstract

The purpose of the present invention is to quantitatively detect a specimen which contains an analyte using a test instrument for immunochromatography. The test instrument for immunochromatography is provided with a referential detection unit for detecting exclusively the activity of a labeled substance that has been captured. Since the referential detection unit certainly detects a signal value corresponding to a maximum capture amount, effects caused by variations in thickness and density of a porous material can be removed by comparing the aforesaid signal value with a signal value detected by a detection unit.

Description

イムノクロマト用検査器具、分析装置、および分析方法Immunochromatographic inspection instrument, analysis apparatus, and analysis method
　本発明は、イムノクロマト用検査器具、分析装置および分析方法に関する。本発明は、特に、血液中に含まれている特定の成分（例えば抗原、抗体、酵素、基質、サイトカインなど）を検出および分析するイムノクロマト用検査器具（チップ）、分析装置、および上記成分を検出および分析する方法に関する。 The present invention relates to an immunochromatographic test instrument, an analysis apparatus, and an analysis method. In particular, the present invention detects an immunochromatographic test instrument (chip) for detecting and analyzing a specific component (for example, antigen, antibody, enzyme, substrate, cytokine, etc.) contained in blood, an analyzer, and the above component And how to analyze.
　イムノクロマト法とは、検出すべき物質を含む検体を抗原抗体反応により簡易的にかつ短時間で検出するための測定法である。そのため、現在多くの場面、例えば病院における臨床検査、研究室における検定試験等に広く使われている。これらの特徴を利用して、イムノクロマト法は妊娠検査薬やインフルエンザ検査薬に用いられており、新たなＰＯＣＴ（Ｐｏｉｎｔ　Ｏｆ　Ｃａｒｅ　Ｔｅｓｔｉｎｇ）の手法として注目を集めている。ここで、ＰＯＣＴとは、患者に出来る限り近い場所で診断するための検査をいう。そのため、ＰＯＣＴによれば、瞬時に提供される検査情報を基に、迅速かつ的確な治療が可能となる。従って、病院での緊急検査や手術中の検査が可能となるので、医療現場におけるニーズが高まっている。 Immunochromatography is a measurement method for detecting a sample containing a substance to be detected simply and in a short time by an antigen-antibody reaction. Therefore, it is now widely used in many scenes, such as clinical examinations in hospitals and certification tests in laboratories. Utilizing these characteristics, immunochromatography is used for pregnancy test drugs and influenza test drugs, and is attracting attention as a new POCT (Point Of Care Testing) method. Here, POCT refers to a test for diagnosis as close as possible to the patient. Therefore, according to POCT, prompt and accurate treatment is possible based on examination information provided instantaneously. Therefore, an emergency inspection at a hospital and an inspection during an operation become possible, and the needs in the medical field are increasing.
　一般的なイムノクロマト法におけるイムノクロマト用検査器具の構造を図４に示す。図４に示すように、検出すべき物質１０５を捕捉するとともに標識物質が結合している第一の捕捉物質１１０（抗体感作ラテックス１０９）が移動可能なように配置されたコンジュゲーションパッド１０８と、検出すべき物質１０５を捕捉する第二の捕捉物質１０４が、多孔体１０１上に固定化された検出部１０３と、液体を吸収する吸収パッド１０７および検体を滴下するサンプルパッド１０２からなる。検出すべき物質１０５が含まれる検体がサンプルパッド１０２に滴下されると、毛細管力によりコンジュゲーションパッド１０８に流れこみ、抗体感作ラテックス１０９に捕捉された後、更に毛管力により多孔体１０１に流れることで第二の捕捉物質１０４に捕捉される。その結果、抗原抗体複合体１１１が形成される。検出は主に目視で行われ、定性的な判定に使用されている。 Fig. 4 shows the structure of an immunochromatographic test instrument in a general immunochromatography method. As shown in FIG. 4, the conjugation pad 108 is arranged so that the first capture substance 110 (antibody-sensitized latex 109) to which the substance 105 to be detected is bound and the label substance is bound can be moved. The second capture substance 104 that captures the substance 105 to be detected includes a detection unit 103 fixed on the porous body 101, an absorption pad 107 that absorbs liquid, and a sample pad 102 that drops a specimen. When a specimen containing the substance 105 to be detected is dropped onto the sample pad 102, it flows into the conjugation pad 108 by capillary force, is captured by the antibody-sensitized latex 109, and further flows into the porous body 101 by capillary force. Thus, the second trapping substance 104 is trapped. As a result, an antigen-antibody complex 111 is formed. Detection is mainly performed visually and is used for qualitative determination.
　近年、イムノクロマト法を定量的に判定する器具や装置に関しても下記の文献に記されている。特許文献１には複数の検出部を設けることが開示されている。これを用いて、反応後の呈色した検出部の数から半定量を行うことが開示されている。特許文献２には検出すべき物質とは無関係の標識抗体と特異的に結合する抗体を固定化する部位を設け、測定時毎回一定の信号値となるように調製した標識抗体を配置することが開示されている。これを用いて、反応後の検出すべき物質とは無関係の標識抗体と特異的に結合する抗体を固定化する部位の信号値と検出部の信号値を比較することで定量することが開示されている。特許文献３には粒子をあらかじめ直接固定化した標準呈色領域を設けることが開示されている。これを用いて、反応後の標準呈色領域と検出部を比較することで定量することが開示されている。特許文献４には、検出部の長さを大きく取ることが開示されている。これを用いて、反応後の呈色領域の長さを読み取ることで定量することが開示されている。特許文献５には温度制御部を設けるシステムを開示している。温度により反応性が変化するため、温度を一定に保つことで定量性を高めることが開示されている。 In recent years, instruments and devices for quantitatively determining immunochromatography have also been described in the following literature. Patent Document 1 discloses providing a plurality of detection units. Using this, it is disclosed to perform semi-quantification from the number of colored detection parts after reaction. In Patent Document 2, a site for immobilizing an antibody that specifically binds to a labeled antibody that is irrelevant to a substance to be detected is provided, and a labeled antibody prepared so that a constant signal value is obtained every time measurement is performed. It is disclosed. Using this, it is disclosed that quantification is performed by comparing the signal value of the site where the antibody that specifically binds to the labeled antibody unrelated to the substance to be detected after the reaction is immobilized with the signal value of the detection unit. ing. Patent Document 3 discloses providing a standard color region in which particles are directly fixed in advance. Using this, it is disclosed that quantification is performed by comparing a standard color region after reaction with a detection unit. Patent Document 4 discloses that the length of the detection unit is increased. Using this, it is disclosed that it quantifies by reading the length of the colored region after reaction. Patent Document 5 discloses a system provided with a temperature control unit. Since the reactivity changes depending on the temperature, it is disclosed that the quantitativeness is improved by keeping the temperature constant.
日本国公開特許公報「特開平０５－００５７４３号公報（１９９３年１月１４日公開）」Japanese Patent Publication “Japanese Patent Laid-Open No. 05-005743” (published on January 14, 1993) 日本国公開特許公報「特開２００１－８３１５３号公報（２００１年３月３０日公開）」Japanese Patent Publication “Japanese Patent Laid-Open No. 2001-83153 (published on March 30, 2001)” 日本国公開特許公報「国際公開第２００６/０８０４３８号（２００６年８月３日公開）」Japanese Patent Publication “International Publication No. 2006/080438 (Released on August 3, 2006)” 日本国公開特許公報「特開２０１０－０２５８８７号公報（２０１０年２月４日公開）」Japanese Patent Publication “Japanese Patent Laid-Open No. 2010-025887 (published on February 4, 2010)” 日本国公開特許公報「特開２０１１－１２８０３８号公報（２０１１年６月３０日公開）」Japanese Patent Publication “JP 2011-128038 A” (published on June 30, 2011)
　イムノクロマト用検査器具自身の問題点として、多孔体自身の密度、厚さが使用する部位で厳密に一定ではないため、第二の捕捉物質の量が器具間で一定にならないといった問題点が挙げられる。具体的には、第二の捕捉物質自身の量および密度が器具間で異なるため、呈色度合いが厳密に一定とならない。 As a problem of the immunochromatography inspection instrument itself, the density and thickness of the porous body itself are not strictly constant at the site to be used, so that the amount of the second capture substance is not constant between instruments. . Specifically, since the amount and density of the second trapping substance itself vary between instruments, the degree of coloration is not strictly constant.
　特許文献２および３に関しては、明記されていないが多孔体自身の密度、厚さを考慮できる可能性がある。しかしながら、特許文献２および３に記載された器具は作製が容易ではなく煩雑である。例えば、特許文献２に記載のように検体とは無関係の標識抗体と特異的に結合する抗体を用いるとすれば、本来検体を検出するための抗体の反応性を阻害しない物質でかつ、バックグラウンド要素にならない材料でなければならない。特許文献３は材料の問題は同様であるがその他にも呈色度合いを一定に保つ工夫が必要となる。 Regarding Patent Documents 2 and 3, although not specified, there is a possibility that the density and thickness of the porous body itself can be considered. However, the devices described in Patent Documents 2 and 3 are not easy to manufacture and are complicated. For example, if an antibody that specifically binds to a labeled antibody unrelated to the specimen is used as described in Patent Document 2, it is a substance that does not inhibit the reactivity of the antibody for detecting the specimen, and the background. The material must not be an element. In Patent Document 3, the problem of the material is the same, but in addition, a device for keeping the coloring degree constant is required.
　その他、特許文献１～４に記載された器具では特許文献５に記載されているように温度による影響により定量性が低くなる。また特許文献５のように温度制御部をもつシステムは簡易的には構成できない。 In addition, the instruments described in Patent Documents 1 to 4 have low quantitativeness due to the influence of temperature as described in Patent Document 5. Further, a system having a temperature control unit as in Patent Document 5 cannot be simply configured.
　本発明は、上記の問題点に鑑みてなされたものであり、その目的は、イムノクロマト法を使用した器具を用いて簡易的に安定な定量値を測定することができるイムノクロマト用検査器具、分析装置、および分析方法を提供することにある。 The present invention has been made in view of the above-described problems, and an object of the present invention is to provide an immunochromatographic test instrument and analyzer capable of simply measuring a stable quantitative value using an instrument using an immunochromatography method. And providing an analysis method.
　上記の課題を解決するために、本発明は、検体が毛管現象により移動可能な多孔体を有するイムノクロマト用検査器具において、検出すべき物質を捕捉するとともに標識物質が結合している第一の捕捉物質が移動可能なように配置された領域と、上記検出すべき物質を特異的に捕捉する第二の捕捉物質が、上記多孔体上に固定された状態で配置された検出部と、上記多孔体上に設けられているとともに、上記第一の捕捉物質を捕捉するための第三の捕捉物質が配置されている参照検出部と、が設けられており、上記第三の捕捉物質は、飽和吸着にて上記第一の捕捉物質を捕捉することを特徴としている。 In order to solve the above-described problems, the present invention provides a first capture in which a substance to be detected is captured and a labeling substance is bound in an immunochromatographic test instrument having a porous body in which a specimen can move by capillary action. An area where the substance is movable, a detection part where the second capture substance that specifically captures the substance to be detected is fixed on the porous body, and the porous And a reference detection unit in which a third capture substance for capturing the first capture substance is disposed, and the third capture substance is saturated. The first capturing substance is captured by adsorption.
　換言すれば、上記構成では、上記第三の捕捉物質が、当該第三の捕捉物質が吸着し得る最大量の第一の捕捉物質を捕捉するように、上記多孔体に対して第一の捕捉物質が流されている。ここで、「飽和吸着」とは、さらに第一の捕捉物質が導入されても、第三の捕捉物質が第一の捕捉物質と吸着しない状態を意味する。 In other words, in the above configuration, the third capture substance captures the first capture substance with respect to the porous body so as to capture the maximum amount of the first capture substance that can be adsorbed by the third capture substance. Material is being washed away. Here, “saturated adsorption” means a state where the third capture substance does not adsorb to the first capture substance even if the first capture substance is further introduced.
　検出部の信号値は、検体中に含まれる対象物質（検出すべき物質）の濃度により値が上下する以外に、多孔体の厚さや密度により信号値の差異を生じる。また、保存環境や使用環境によっても信号値が変化する。上記構成であれば、参照検出部の信号値は、必ず飽和吸着で捕捉された第一の捕捉物質の量（常に一定の量）に相当する信号値が検出されるため、参照検出部の信号値を参照することで上記要因の影響を排除することが可能になる。 The signal value of the detection unit varies depending on the thickness and density of the porous body, as well as the value varies depending on the concentration of the target substance (substance to be detected) contained in the specimen. In addition, the signal value varies depending on the storage environment and use environment. With the above configuration, since the signal value of the reference detection unit always detects a signal value corresponding to the amount of the first capture substance captured by saturation adsorption (always a constant amount), the signal of the reference detection unit It is possible to eliminate the influence of the above factors by referring to the values.
　例えば、多孔体の材料と種類、および規格が同じであっても、局所的な厚さや密度が異なることで信号値が変化してしまう。また、標識物質が発する信号（例えば、蛍光）は、様々な条件によって変化する。例えば、標識物質が古くなれば、当該標識物質が発する信号は、時間が経つのに伴って弱くなる。これらの理由により、一定量の標識物質が発する信号は、常に一定であるわけではない。このとき、上記構成であれば、第三の捕捉物質が常に一定量の第一の捕捉物質を結合することができるので、例え標識物質が古くなっていたとしても、検出を行う時点における、一定量の第一の捕捉物質あたりの信号（例えば、蛍光の強度）を検出することができる。そして、当該信号（一定量の第一の捕捉物質あたりの信号）に基づいて、上記検出部にて検出される信号を補正することによって、上記検出部で捕捉されている第一の捕捉物質の量を正確に算出することができる。第一の捕捉物質は、検出すべき物質を捕捉するものなので、検出部に捕捉されている第一の捕捉物質の量を正確に算出することができれば、検出部に捕捉されている検出すべき物質の量も正確に算出することができる。 For example, even if the material, type, and standard of the porous body are the same, the signal value changes due to different local thickness and density. Moreover, the signal (for example, fluorescence) which a labeling substance emits changes with various conditions. For example, if the labeling substance becomes old, the signal emitted by the labeling substance becomes weaker with time. For these reasons, the signal emitted by a certain amount of labeling substance is not always constant. At this time, with the above configuration, since the third capture substance can always bind a certain amount of the first capture substance, even if the labeling substance is old, it is constant at the time of detection. A signal (eg, fluorescence intensity) per amount of the first capture material can be detected. And based on the said signal (signal per fixed amount of 1st capture | acquisition substances), by correcting the signal detected by the said detection part, of the 1st capture | acquisition substance captured by the said detection part The amount can be calculated accurately. Since the first capture substance captures the substance to be detected, if the amount of the first capture substance captured by the detection unit can be accurately calculated, the detection should be detected by the detection unit. The amount of substance can also be calculated accurately.
　よって、上記構成によれば、イムノクロマト法を使用した器具を用いて簡易的に安定な定量値を測定することができるイムノクロマト用検査器具を提供することができる。 Therefore, according to the above configuration, it is possible to provide an immunochromatographic test instrument that can easily measure a stable quantitative value using an instrument using an immunochromatography method.
　本発明に係るイムノクロマト用検査器具では、上記参照検出部は、上記検出部の下流側に設けられていることが好ましい。ここで、「下流」とは、検体が毛管現象により移動する流れの下流である。 In the immunochromatographic test instrument according to the present invention, it is preferable that the reference detection unit is provided on the downstream side of the detection unit. Here, “downstream” is downstream of the flow in which the specimen moves by capillary action.
　上記構成であれば、第一の捕捉物質は検出部を通過した後、参照検出部に送液されるため、検出部の反応（第二の捕捉物質による、検出すべき物質の捕捉）を優先させることが可能となる。また、対象物質も参照検出部を通過する前に検出部で反応させることができるため、参照検出部に対する非特異吸着などの影響を少なくすることができる。 If it is the said structure, since a 1st capture substance passes through a detection part and is sent to a reference detection part, it will give priority to reaction of a detection part (capturing of the substance which should be detected by a 2nd capture substance) It becomes possible to make it. In addition, since the target substance can also be reacted at the detection unit before passing through the reference detection unit, the influence of non-specific adsorption or the like on the reference detection unit can be reduced.
　本発明に係るイムノクロマト用検査器具では、上記多孔体に流される上記第一の捕捉物質の量および上記第一の捕捉物質に結合している標識物質の量は、いずれも上記第二の捕捉物質の量と上記第三の捕捉物質の量との和以上の量であることが好ましい。 In the immunochromatographic test instrument according to the present invention, the amount of the first capture substance flowing through the porous body and the amount of the labeling substance bound to the first capture substance are both the second capture substance. It is preferable that the amount is equal to or greater than the sum of the amount of the third trapping substance and the amount of the third trapping substance.
　上記構成であれば、第一の捕捉物質の量は、参照検出部において必ず飽和吸着になるように捕捉される条件が成り立つため、第一の捕捉物質の量の調整を簡易的に行うことができる。また、上記構成によれば、より正確に、対象物質を測定することができる。 With the above configuration, the amount of the first trapping substance can be easily adjusted because the amount of the first trapping substance is satisfied so that the reference detection unit always captures saturated adsorption. it can. Moreover, according to the said structure, a target substance can be measured more correctly.
　本発明に係るイムノクロマト用検査器具では、上記多孔体は、分岐しており、上記分岐した多孔体の１つに、上記検出部が設けられており、上記分岐した多孔体のうち、上記検出部が設けられた多孔体とは別の多孔体に、上記参照検出部が設けられていることが好ましい。 In the immunochromatographic test instrument according to the present invention, the porous body is branched, and the detection section is provided in one of the branched porous bodies. Among the branched porous bodies, the detection section is provided. It is preferable that the reference detection unit is provided in a porous body different from the porous body provided with.
　上記構成であれば、参照検出部と検出部とがそれぞれ独立して反応できるため、お互いに影響することなく検出することが可能となる。 If it is the said structure, since a reference detection part and a detection part can each react independently, it becomes possible to detect without affecting each other.
　本発明に係るイムノクロマト用検査器具では、上記多孔体に流される上記第一の捕捉物質の量および上記第一の捕捉物質に結合している標識物質の量は、それぞれ上記検出部における上記第二の捕捉物質の量または上記参照検出部における上記第三の捕捉物質の量の２倍以上であることが好ましい。特に、上記多孔体に流される上記第一の捕捉物質の量および上記第一の捕捉物質に結合している標識物質の量はそれぞれ、上記検出部における上記第二の捕捉物質の量及び上記参照検出部における上記第三の捕捉物質の量のうち相対的に多い量の２倍以上であることが好ましい。 In the immunochromatographic test instrument according to the present invention, the amount of the first capture substance flowing into the porous body and the amount of the labeling substance bound to the first capture substance are respectively determined by the second detection unit. It is preferable that the amount is not less than twice the amount of the capture substance or the amount of the third capture substance in the reference detection unit. In particular, the amount of the first capture substance flowing into the porous body and the amount of the labeling substance bound to the first capture substance are the amount of the second capture substance in the detection unit and the reference, respectively. It is preferable that the amount is more than twice the relatively large amount of the third capturing substance in the detection unit.
　上記構成であれば、第一の捕捉物質の量は参照検出部において必ず飽和吸着になるように捕捉される条件が成り立つため、第一の捕捉物質の量の調整を簡易的に行うことができる。また、上記構成によれば、より正確に対象物質を測定することができる。 With the above configuration, the amount of the first capture substance can be easily adjusted because the amount of the first capture substance is satisfied so that the reference detection unit always captures the saturated adsorption. . Moreover, according to the said structure, a target substance can be measured more correctly.
　本発明に係るイムノクロマト用検査器具では、上記参照検出部および上記検出部は、同一の検出手段を備えていることが好ましい。 In the immunochromatographic test instrument according to the present invention, the reference detection unit and the detection unit preferably include the same detection means.
　上記構成であれば、同一の検出手段で検出するため、例えば温度などの使用環境の変化により検出器自身が影響を受けても、検出部および参照検出部における信号値の変動幅は同一であるので、その影響を排除することができる。 In the case of the above configuration, since the detection is performed by the same detection means, even if the detector itself is affected by a change in the usage environment such as temperature, the fluctuation range of the signal value in the detection unit and the reference detection unit is the same. Therefore, the influence can be eliminated.
　本発明に係るイムノクロマト用検査器具では、上記参照検出部および上記検出部は、外部から色の濃淡または発光を観察できる構造であることが好ましい。 In the immunochromatographic test instrument according to the present invention, the reference detection unit and the detection unit preferably have a structure capable of observing color shading or light emission from the outside.
　上記構成であれば、色の濃淡または発光に基づいて、対象物質を光学的に検出することが可能となる。光学的に検出される対象物質は、それ自身が発色または発光するものであってもよいし、発色または発光する物質で修飾されたものであってもよい。対象物質を色の濃淡に基づいて検出する方法としては、発色する物質による色の濃淡を光学的に検出すればよい。また、対象物質を発光に基づいて検出する方法としては、発光物質による発光を光学的に検出すればよい。なお、発光には化学反応発光及び蛍光（光励起による発光）などが包含される。 With the above configuration, the target substance can be optically detected based on color shading or light emission. The target substance to be detected optically may be one that develops color or emits light, or may be one that is modified with a substance that produces or emits light. As a method for detecting the target substance based on the color shading, the shading of the color due to the coloring substance may be optically detected. In addition, as a method for detecting the target substance based on light emission, light emission by the light emitting substance may be optically detected. The light emission includes chemical reaction light emission and fluorescence (light emission by photoexcitation).
　本発明に係るイムノクロマト用検査器具では、上記参照検出部および上記検出部は、それぞれ作用電極および参照電極を備えていることが好ましい。 In the immunochromatographic test instrument according to the present invention, the reference detection unit and the detection unit preferably include a working electrode and a reference electrode, respectively.
　上記構成を用いれば、検出部において、対象物質を電気化学的に検出することが可能となる。電気化学的に検出される対象物質は、それ自身が電気化学的に活性なものであってもよいし、電気化学的に活性な物質で修飾されたものであってもよい。対象物質を電気化学的に検出する方法としては、電気化学的に活性な物質から得られる電流値を検出電極にて測定すればよい。 If the above configuration is used, the target substance can be detected electrochemically in the detection unit. The target substance to be detected electrochemically may be one that is electrochemically active per se, or one that is modified with an electrochemically active substance. As a method for electrochemically detecting a target substance, a current value obtained from an electrochemically active substance may be measured with a detection electrode.
　本発明に係るイムノクロマト用検査器具では、上記作用電極および上記参照電極は、大きさが同一であることが好ましい。 In the immunochromatographic test instrument according to the present invention, it is preferable that the working electrode and the reference electrode have the same size.
　上記構成であれば、同一の大きさの電極を用いるので、電極界面に拡がる拡散層の大きさを考慮しなくとも、簡易に信号値同士を比較することが可能となる。 In the above configuration, since the electrodes having the same size are used, it is possible to easily compare the signal values without considering the size of the diffusion layer spreading on the electrode interface.
　本発明に係るイムノクロマト用検査器具では、上記第一の捕捉物質が、上記検出すべき物質に対する抗体であることが好ましい。 In the immunochromatographic test instrument according to the present invention, the first capture substance is preferably an antibody against the substance to be detected.
　生化学的な分析に用いられるイムノクロマト用検査器具の分析装置の検出対象となる物質は、生体内タンパク質であることが多い。変性しにくい抗体は、捕捉物質として使用するには最適な物質である。 In many cases, a substance to be detected by an analyzer of an immunochromatographic test instrument used for biochemical analysis is an in vivo protein. Antibodies that are difficult to denature are optimal substances for use as capture substances.
　本発明に係るイムノクロマト用検査器具では、上記第二の捕捉物質が、上記検出すべき物質に対する抗体であることが好ましい。 In the immunochromatographic test instrument according to the present invention, the second capture substance is preferably an antibody against the substance to be detected.
　生化学的な分析に用いられるイムノクロマト用検査器具の分析装置の検出対象となる物質は、生体内タンパク質であることが多い。変性しにくい抗体は、捕捉物質として使用するには最適な物質である。 In many cases, a substance to be detected by an analyzer of an immunochromatographic test instrument used for biochemical analysis is an in vivo protein. Antibodies that are difficult to denature are optimal substances for use as capture substances.
　本発明に係るイムノクロマト用検査器具では、上記第三の捕捉物質が、上記第一の捕捉物質に対する抗体であることが好ましい。 In the immunochromatographic test instrument according to the present invention, the third capture substance is preferably an antibody against the first capture substance.
　生化学的な分析に用いられるイムノクロマト用検査器具の分析装置の検出対象となる物質は、生体内タンパク質であることが多い。変性しにくい抗体は、捕捉物質として使用するには最適な物質である。 In many cases, a substance to be detected by an analyzer of an immunochromatographic test instrument used for biochemical analysis is an in vivo protein. Antibodies that are difficult to denature are optimal substances for use as capture substances.
　本発明に係るイムノクロマト用検査器具では、上記標識物質が、微粒子であることが好ましい。 In the immunochromatographic test instrument according to the present invention, the labeling substance is preferably a fine particle.
　上記構成であれば、微粒子を標識物質として使用するため、第一の捕捉物質の量を微粒子の量として検出することができる。微粒子の量は例えば、色の濃淡で検出することが可能である。 With the above configuration, since the fine particles are used as the labeling substance, the amount of the first capture substance can be detected as the amount of the fine particles. The amount of fine particles can be detected by, for example, color shading.
　本発明に係るイムノクロマト用検査器具では、上記標識物質が、蛍光材料であることが好ましい。 In the immunochromatographic test instrument according to the present invention, the labeling substance is preferably a fluorescent material.
　上記構成であれば、蛍光材料を標識物質として使用するため、直接蛍光検出することで簡便に検出することが可能となる。 With the above configuration, since a fluorescent material is used as a labeling substance, it can be easily detected by direct fluorescence detection.
　本発明に係るイムノクロマト用検査器具では、上記標識物質が、電気化学的活性物質であることが好ましい。 In the immunochromatographic test instrument according to the present invention, the labeling substance is preferably an electrochemically active substance.
　上記構成であれば電気化学的活性物質を標識材料として使用するため、直接電気化学的に検出することで簡便に検出することが可能となる。 In the above configuration, since the electrochemically active substance is used as the labeling material, it can be easily detected by direct electrochemical detection.
　本発明に係るイムノクロマト用検査器具では、上記参照検出部の下流に、上記第一の捕捉物質を捕捉するための第四の捕捉物質が配置されている判定部が設けられていることが好ましい。 In the immunochromatographic test instrument according to the present invention, it is preferable that a determination unit in which a fourth capture substance for capturing the first capture substance is disposed downstream of the reference detection unit.
　上記構成であれば検出するための反応が完了したか否か判定することが可能となる。 If it is the above-mentioned composition, it will become possible to judge whether the reaction for detection was completed.
　本発明に係るイムノクロマト用検査器具では、上記検出部の信号値および上記参照検出部の信号値から導出される検出信号値と、上記検出すべき物質の濃度との相関関係に関する情報が書き込まれたバーコードまたはＩＣタグが設けられていることが好ましい。 In the immunochromatographic test instrument according to the present invention, information on the correlation between the detection signal value derived from the signal value of the detection unit and the signal value of the reference detection unit and the concentration of the substance to be detected is written. A bar code or IC tag is preferably provided.
　上記構成であれば、チップに予め情報を書き込んでおくことができる。なお、製造時のロットや試薬のロットに応じ、書き込む情報を変えることが可能となる。 With the above configuration, information can be written in advance on the chip. The information to be written can be changed according to the lot at the time of manufacture and the lot of the reagent.
　本発明に係る分析装置は、上記課題を解決するために、本発明に係るイムノクロマト用検査器具における上記標識物質が発する信号を検出する検出器を備えていることを特徴としている。 In order to solve the above-described problems, the analyzer according to the present invention is characterized by including a detector that detects a signal emitted from the labeling substance in the immunochromatographic test instrument according to the present invention.
　上記構成であれば、参照検出部の信号値は、必ず飽和吸着になるように捕捉された第一の捕捉物質に相当する値が検出されるため、参照検出部の信号値を参照することで、使用環境などの影響を排除することが可能となる。 If it is the said structure, since the value corresponding to the 1st capture | acquisition substance capture | acquired so that it may become saturation adsorption | suction is surely detected for the signal value of a reference detection part, by referring the signal value of a reference detection part, It becomes possible to eliminate the influence of the usage environment.
　本発明に係る分析装置では、上記検出部の信号値と上記参照検出部の信号値とを比較した相対信号値から、上記検出すべき物質の濃度を算出することが好ましい。本発明に係る分析装置においては、演算部が上記検出部の信号値と上記参照検出部の信号値とを比較した相対信号値から、上記検出すべき物質の濃度を算出する。 In the analyzer according to the present invention, it is preferable to calculate the concentration of the substance to be detected from a relative signal value obtained by comparing the signal value of the detection unit and the signal value of the reference detection unit. In the analyzer according to the present invention, the calculation unit calculates the concentration of the substance to be detected from the relative signal value obtained by comparing the signal value of the detection unit and the signal value of the reference detection unit.
　上記構成であれば、チップ製造後の時間経過や保存状態、使用環境による影響を、例えば参照検出部の信号値との比をとり相殺することで、対象物質の濃度に依存した信号値のみを得ることが可能となる。この他にも差や補正係数を掛けるなど補正手段は問わない。 With the above configuration, only the signal value depending on the concentration of the target substance can be obtained by offsetting the influence of the elapsed time, storage state, and usage environment after chip manufacture, for example, by taking the ratio with the signal value of the reference detection unit. Can be obtained. In addition, any correction means such as a difference or a correction coefficient is applied.
　本発明に係る分析装置は、上記参照検出部の信号値が基準値以下である場合に、警告を発することが好ましい。本発明に係る分析装置においては、警告発生部が、上記参照検出部の信号値が基準値以下である場合に、警告を発する。 The analyzer according to the present invention preferably issues a warning when the signal value of the reference detection unit is equal to or less than a reference value. In the analyzer according to the present invention, the warning generation unit issues a warning when the signal value of the reference detection unit is equal to or less than a reference value.
　上記構成であれば、信頼の無いデータをユーザーに表示することなく、警告を発することで、ユーザーに検出エラーが生じたことを伝えることができる。 With the above configuration, it is possible to inform the user that a detection error has occurred by issuing a warning without displaying unreliable data to the user.
　本発明に係る分析装置は、上記検出部の信号値および上記参照検出部の信号値について、検出する検出器は同じであることが好ましい。 In the analyzer according to the present invention, it is preferable that the detector to detect the signal value of the detection unit and the signal value of the reference detection unit is the same.
　上記構成であれば、同一の検出手段を用いるため、上記検出部の信号値および上記参照検出部の信号値に対して、共通の検出器を用いることができ、分析装置の構成を簡易化することができる。 With the above configuration, since the same detection means is used, a common detector can be used for the signal value of the detection unit and the signal value of the reference detection unit, thereby simplifying the configuration of the analyzer. be able to.
　本発明に係る分析装置は、バーコードまたはＩＣタグ用のリーダーが設けられていることが好ましい。 The analyzer according to the present invention is preferably provided with a reader for barcode or IC tag.
　上記構成であれば、イムノクロマト用検査器具に設けられたバーコードまたはＩＣタグから情報を読み取ることが可能となる。そして、読み取った情報を校正などに使用できる。 If it is the said structure, it will become possible to read information from the barcode provided in the inspection instrument for immunochromatography or an IC tag. The read information can be used for calibration.
　本発明に係る分析方法は、上記課題を解決するために、本発明のイムノクロマト用検査器具を用い、上記参照検出部の信号値と上記検出部の信号値とを比較して、検出信号値を算出することを特徴としている。 In order to solve the above problems, the analysis method according to the present invention uses the immunochromatographic test instrument of the present invention, compares the signal value of the reference detection unit and the signal value of the detection unit, and determines the detection signal value. It is characterized by calculating.
　上記構成であれば、チップ製造後の時間経過や保存状態、使用環境による影響を、参照検出部の信号値と検出部の信号値と比較した相対信号値から、上記検出すべき物質の濃度を算出することが可能である。 If it is the said structure, the density | concentration of the said substance which should be detected from the relative signal value which compared the signal value of a reference detection part with the signal value of a reference detection part and the signal value of a detection part about the time passage after a chip manufacture, a storage state, and a use environment. It is possible to calculate.
　本発明に係る分析方法では、上記検出信号値は、上記検出部の信号値と上記参照検出部の信号値との比から算出されることが好ましい。 In the analysis method according to the present invention, it is preferable that the detection signal value is calculated from a ratio between the signal value of the detection unit and the signal value of the reference detection unit.
　上記構成であれば、チップ製造後の時間経過や保存状態、使用環境による影響を、参照検出部の信号値と検出部の信号値との比をとり相殺することで、対象物質の濃度に依存した信号値のみを得ることが可能となる。 With the above configuration, it depends on the concentration of the target substance by offsetting the influence of the elapsed time, storage state, and usage environment after chip manufacture by taking the ratio of the signal value of the reference detector to the signal value of the detector. Only the obtained signal value can be obtained.
　本発明に係る分析方法では、上記参照検出部の信号値が基準値以下である場合に、検出エラーと判断することが好ましい。 In the analysis method according to the present invention, it is preferable to determine a detection error when the signal value of the reference detection unit is equal to or less than a reference value.
　上記構成であれば、信頼の無いデータをユーザーに表示することなく、ユーザーに検出エラーを伝えることができる。 With the above configuration, a detection error can be transmitted to the user without displaying unreliable data to the user.
　本発明に係る分析方法では、上記検出信号値と上記検出すべき物質の濃度との相関関係を予め上記イムノクロマト用検査器具へ入力し、上記検出信号値と上記相関関係とから、上記検出すべき物質の濃度を算出することが好ましい。 In the analysis method according to the present invention, the correlation between the detection signal value and the concentration of the substance to be detected is previously input to the immunochromatographic test instrument, and the detection should be performed from the detection signal value and the correlation. It is preferable to calculate the concentration of the substance.
　上記構成であれば、参照検出部と検出部とを用いて検出信号値を算出するため、実際の値に近い値を出力することが可能となる。 With the above configuration, since the detection signal value is calculated using the reference detection unit and the detection unit, a value close to the actual value can be output.
　本発明に係る分析方法では、上記イムノクロマト用検査器具の製造ロットおよび試薬ロットの少なくとも一方に関する情報に基づいて、上記相関関係を補正することが好ましい。 In the analysis method according to the present invention, it is preferable to correct the correlation based on information relating to at least one of a production lot and a reagent lot of the immunochromatographic test instrument.
　上記構成であれば、製造ロットおよび／または試薬ロットの差異により生じた上記相関関係の差異を補正することが可能となる。 With the above configuration, it is possible to correct the difference in the correlation caused by the difference in the production lot and / or the reagent lot.
　本発明に係る分析方法では、上記情報を、上記イムノクロマト用検査器具に設けられたバーコードまたはＩＣタグから読み取ることが好ましい。 In the analysis method according to the present invention, it is preferable that the information is read from a barcode or an IC tag provided on the immunochromatographic test instrument.
　上記構成であれば、ユーザーに手間をかけさせること無く、簡便に補正を行うことが可能となる。 If it is the above-mentioned composition, it will become possible to carry out amendment simply without making a user troublesome.
　本発明によれば、使用する多孔体自身の不均一さに依存することなく、定量的な検出がイムノクロマト用検査器具で可能となる。また、保存状態、使用するまでの期間または使用環境に依存することなく、本来の正確な値を検出することができる。 According to the present invention, quantitative detection is possible with an immunochromatographic test instrument without depending on the non-uniformity of the porous body itself used. In addition, the original accurate value can be detected without depending on the storage state, the period until use or the use environment.
　本発明によれば、簡便な構成を有する素子または装置によって、本来の正確な値を検出することができる。 According to the present invention, the original accurate value can be detected by an element or device having a simple configuration.
本発明の実施の形態１に係るイムノクロマト用検査器具の概略図である。It is the schematic of the test | inspection instrument for immunochromatography which concerns on Embodiment 1 of this invention. 本発明の実施の形態１に係るイムノクロマト用検査器具の概略図である。It is the schematic of the test | inspection instrument for immunochromatography which concerns on Embodiment 1 of this invention. 本発明の実施の形態２に係るイムノクロマト用検査器具の概略図である。It is the schematic of the test | inspection instrument for immunochromatography which concerns on Embodiment 2 of this invention. 従来のイムノクロマト用検査器具を示す概略図である。It is the schematic which shows the conventional test | inspection instrument for immunochromatography.
　以下、本発明に係る分析装置の実施形態について、図面を参照しながら説明するが、本発明は、これらに限定されない。 Hereinafter, embodiments of the analysis apparatus according to the present invention will be described with reference to the drawings, but the present invention is not limited thereto.
　本明細書において、用語「サンプル」とは、イムノクロマト用検査器具に導入される検体（被検物）をいい、検出の対象としている目的物質（検出すべき物質）を含んでいても、いなくてもよい。 In this specification, the term “sample” refers to a specimen (test object) introduced into an immunochromatographic test instrument, whether or not it includes a target substance (substance to be detected) to be detected. May be.
　本明細書において、用語「捕捉物質」とは、対象物質と特異的に相互作用することによって、この対象物質と共有結合または非共有結合を形成する物質をいう。捕捉物質は、具体的に、対象物質との間でホストとゲストとの関係を有する物質であり、捕捉物質としては、例えば、抗原、抗体、酵素、基質、リガンド、レセプター、ＤＮＡ、糖、ペプチド、合成高分子（例えばモレキュラーインプリントポリマー）などが挙げられる。 In this specification, the term “capture substance” refers to a substance that forms a covalent bond or a non-covalent bond with the target substance by specifically interacting with the target substance. The capture substance is specifically a substance having a relationship between a host and a guest with the target substance. Examples of the capture substance include antigens, antibodies, enzymes, substrates, ligands, receptors, DNA, sugars, peptides And synthetic polymers (for example, molecular imprint polymer).
　本明細書において、用語「上流」および「下流」は、多孔体内における流体の流れを基準とした概念であり、液体が流れていく方向を「下流」、液体の流れと逆方向を上流と呼ぶ。特に説明を加えない限り、多孔体におけるサンプルパッド側が「上流」である。 In this specification, the terms “upstream” and “downstream” are concepts based on the flow of fluid in the porous body, and the direction in which the liquid flows is referred to as “downstream”, and the direction opposite to the flow of the liquid is referred to as upstream. . Unless otherwise specified, the sample pad side in the porous body is “upstream”.
　〔実施の形態１〕
　図１に基づいて、本発明の実施の形態１について説明する。図１は、本発明の実施の形態１に係るイムノクロマト用検査器具の模式図である。 [Embodiment 1]
A first embodiment of the present invention will be described with reference to FIG. FIG. 1 is a schematic diagram of an immunochromatographic test instrument according to Embodiment 1 of the present invention.
　＜１．イムノクロマト用検査器具＞
　本実施形態に係るイムノクロマト用検査器具は、表面に粘着剤が塗られたバッキングシート１５上に吸収パッド７の下端部を多孔体１の上端部と、コンジュゲーションパッド８の上端部と多孔体１の下端部と、サンプルパッド２の上端部をコンジュゲーションパッド８下端部と数ミリ重ね合わせるように貼り付けることで構成される。貼り合わせ後、数１０μｍ～数ｃｍ幅で切断し短冊状とすることができる。好ましくは数百μｍ～数ｍｍ幅が検体量の観点から好ましい。周知の通り、切断した個々の器具をハウジング内に入れることで、ユーザーが使用しやすい形状にすることができる。 <1. Immunochromatographic Inspection Equipment>
In the immunochromatographic test instrument according to the present embodiment, the lower end of the absorbent pad 7 is placed on the backing sheet 15 whose surface is coated with an adhesive, the upper end of the porous body 1, the upper end of the conjugation pad 8 and the porous body 1. And the upper end of the sample pad 2 are pasted so as to overlap with the lower end of the conjugation pad 8 by several millimeters. After pasting, it can be cut into a strip shape by cutting with a width of several tens of μm to several cm. A width of several hundred μm to several mm is preferable from the viewpoint of the amount of specimen. As is well known, each cut instrument can be placed in the housing to make it easy for the user to use.
　バッキングシート１５は各材料を組み合わせるための基板としての機能を有する。吸収パッド７は毛管力により多孔体１内を流れてきた検体を吸収する機能を有する。コンジュゲーションパッド８は標識物質が結合している第一の捕捉物質９（例えば抗体感作ラテックス粒子）が保存された領域であり、液体が通過した際、移動可能となるように保存する。サンプルパッド２は、検体を注入するための領域を指す。 The backing sheet 15 has a function as a substrate for combining the materials. The absorption pad 7 has a function of absorbing the specimen flowing through the porous body 1 by capillary force. The conjugation pad 8 is an area where a first capture substance 9 (for example, antibody-sensitized latex particles) to which a label substance is bound is stored, and is stored so that it can move when a liquid passes through. The sample pad 2 indicates an area for injecting a specimen.
　更に多孔体上には検出部３が設けられている。この検出部３には、検出および分析の対象となる物質を捕捉する第二の捕捉物質４が固定化されている。さらに、多孔体１内には、コンジュゲーションパッド８内に保存された標識された第一の捕捉物質９および標識物質の活性を評価するための参照検出部５が設けられている。この参照検出部５には、第一の捕捉物質を捕捉する第三の捕捉物質６が固定化されている。検出部３と参照検出部５とは、多孔体１内であれば何処に設けられても構わないが、サンプルパッド２側に検出部３、吸収パッド７側に参照検出部５を設けることが反応性の観点から好ましい。 Furthermore, the detection part 3 is provided on the porous body. A second capture substance 4 that captures a substance to be detected and analyzed is immobilized on the detection unit 3. Further, in the porous body 1, a labeled first capture substance 9 stored in the conjugation pad 8 and a reference detection unit 5 for evaluating the activity of the label substance are provided. A third capture substance 6 that captures the first capture substance is immobilized on the reference detection unit 5. The detection unit 3 and the reference detection unit 5 may be provided anywhere within the porous body 1, but the detection unit 3 may be provided on the sample pad 2 side and the reference detection unit 5 may be provided on the absorption pad 7 side. It is preferable from the viewpoint of reactivity.
　以下、本実施形態に係るイムノクロマト用検査器具が備えている部材について詳細に説明する。 Hereinafter, members provided in the immunochromatographic inspection instrument according to this embodiment will be described in detail.
　＜１．１　多孔体＞
　多孔体１は、毛管現象によって液体が移動可能な媒体であればその材質は制限されない。具体的には例えば、セルロース、セルロース誘導体、ニトロセルロース、エチレン酢酸ビニル、ポリウレタン、ポリメチルメタクリレート、ナイロン樹脂、ポリビニリデンジフルオリド（ＰＶＤＦ）等の多孔性合成ポリマー等を原料とする織布、不織布やガラス繊維フィルター、各種の濾紙等が挙げられ、セルロース、セルロース誘導体、多孔性合成ポリマーを用いたものが好ましく、ニトロセルロースを用いたものが特に好ましい。 <1.1 Porous body>
The material of the porous body 1 is not limited as long as the medium can move the liquid by capillary action. Specifically, for example, woven fabrics, nonwoven fabrics, and the like made of porous synthetic polymers such as cellulose, cellulose derivatives, nitrocellulose, ethylene vinyl acetate, polyurethane, polymethyl methacrylate, nylon resin, polyvinylidene difluoride (PVDF), etc. Examples thereof include glass fiber filters and various filter papers, and those using cellulose, cellulose derivatives, and porous synthetic polymers are preferable, and those using nitrocellulose are particularly preferable.
　＜１．２　サンプルパッド＞
　サンプルパッド２は、検体が滴下される部位である。また、検体中に不溶性物質が存在している場合、除去フィルターとしても作用する。サンプルパッド２に使用可能な材料としては、親水性であり、フィルター機能を有するものであれば特に限定されず、例えば、再生セルロース、酢酸セルロース、ニトロセルロース、ポリアクリロニトリル、エチレン酢酸ビニル、ポリウレタン、ポリメチルメタクリレート、ナイロン樹脂、ガラス繊維、パルプ、綿、レーヨン、アクリル、ポリエステルなどが例示される。 <1.2 Sample pad>
The sample pad 2 is a part where the specimen is dropped. Further, when an insoluble substance is present in the specimen, it also functions as a removal filter. The material that can be used for the sample pad 2 is not particularly limited as long as it is hydrophilic and has a filter function. For example, regenerated cellulose, cellulose acetate, nitrocellulose, polyacrylonitrile, ethylene vinyl acetate, polyurethane, poly Examples include methyl methacrylate, nylon resin, glass fiber, pulp, cotton, rayon, acrylic, and polyester.
　＜１．３　検出部＞
　検出部３は、上述したように、多孔体１を流れる流体中の物質を検出する部位であり、図１に示すように、検出部３には、検出および分析の対象となる物質（以下、対象物質とも称する。）を捕捉する第二の捕捉物質４が固定化されている。第二の捕捉物質４は、対象物質とホスト－ゲストの関係にある物質（例えば、抗体、抗原、ペプチド、ＤＮＡ、糖、合成高分子（例えばモレキュラーインプリントポリマー）、酵素、基質、リガンドまたはレセプターなど）であればよく、特に、抗体または合成高分子は、活性が安定しているので好ましい。また、第二の捕捉物質４を固定化する方法としては、物理的吸着法、化学結合法または共有結合法などの公知の方法が適宜採用され得る。また固定化するためにスポッターを用いることが好ましい。特にラインプロットを行うことでライン状に抗体溶液を定量的に滴下することが可能となる。固定化後、風乾および熱風乾燥などにより、多孔体１を乾燥させる。 <1.3 Detection unit>
As described above, the detection unit 3 is a part that detects a substance in the fluid flowing through the porous body 1. As shown in FIG. 1, the detection unit 3 includes a substance (hereinafter, referred to as a target of detection and analysis). A second capture substance 4 that captures the target substance is immobilized. The second capture substance 4 is a substance (for example, an antibody, an antigen, a peptide, DNA, a sugar, a synthetic polymer (for example, a molecular imprint polymer), an enzyme, a substrate, a ligand, or a receptor that has a host-guest relationship with the target substance. In particular, an antibody or a synthetic polymer is preferable because its activity is stable. In addition, as a method for immobilizing the second capture substance 4, a known method such as a physical adsorption method, a chemical bonding method, or a covalent bonding method can be appropriately employed. Moreover, it is preferable to use a spotter for immobilization. In particular, by performing a line plot, it is possible to quantitatively drop the antibody solution in a line shape. After immobilization, the porous body 1 is dried by air drying or hot air drying.
　検出部３の構成は、特に限定されず、対象物質の検出方法によって適宜決定され得る。例えば色の濃淡または発光（蛍光を含む）などに基づいて、対象物質を光学的に検出する場合、外部から色を観察できるような構造とすればよい。例えば、イムノクロマト用検査器具をハウジングした場合でも、観察窓を設けることで検出が可能となる。 The configuration of the detection unit 3 is not particularly limited, and can be appropriately determined depending on the detection method of the target substance. For example, when the target substance is optically detected based on color shading or light emission (including fluorescence), the structure may be such that the color can be observed from the outside. For example, even when an immunochromatographic inspection instrument is housed, detection is possible by providing an observation window.
　また、対象物質を電気化学的に検出する場合、検出部３は、多孔体１の上部または下部に形成された検出電極を有する検出手段を備えていればよい。検出電極は、少なくとも参照電極および作用電極の２電極から構成されていればよいが、参照電極および作用電極に加えて対向電極を備えている３電極から構成されていることが好ましい。３電極であれば、基準電位が取れるためより正確な測定を行うことができる。 In addition, when the target substance is detected electrochemically, the detection unit 3 only needs to include detection means having a detection electrode formed on the top or bottom of the porous body 1. The detection electrode only needs to be composed of at least two electrodes, that is, a reference electrode and a working electrode, but is preferably composed of three electrodes provided with a counter electrode in addition to the reference electrode and the working electrode. If there are three electrodes, the reference potential can be taken, so that more accurate measurement can be performed.
　参照電極、作用電極および対向電極は、従来のフォトリソグラフィ技術を利用した微細加工技術によって形成することができる。電極の導電性材料として、例えば金、白金、銀、クロム、チタン、イリジウム、銅またはカーボンなどを用いることができる。参照電極には、基準電位の安定性の観点から、銀／塩化銀電極を用いることが好ましい。 The reference electrode, the working electrode, and the counter electrode can be formed by a microfabrication technique using a conventional photolithography technique. As the conductive material of the electrode, for example, gold, platinum, silver, chromium, titanium, iridium, copper, or carbon can be used. As the reference electrode, a silver / silver chloride electrode is preferably used from the viewpoint of stability of the standard potential.
　また、参照電極、作用電極および対向電極は、大きさが同一であることが好ましい。上記構成によれば、電極界面に拡がる拡散層の大きさを考慮せずに、信号値を比較することができる。 In addition, it is preferable that the reference electrode, the working electrode, and the counter electrode have the same size. According to the above configuration, the signal values can be compared without considering the size of the diffusion layer spreading on the electrode interface.
　＜１．４　参照検出部＞
　参照検出部５は、上述したように、チップ内に保存された標識された第一の捕捉物質９および標識物質の活性を評価する部位である。図１に示すように、検出部３と吸収パッド７との間に設けられた参照検出部５には、第一の捕捉物質を捕捉するための第三の捕捉物質６が固定化されている。 <1.4 Reference detection unit>
As described above, the reference detection unit 5 is a site for evaluating the activity of the labeled first capture substance 9 and the label substance stored in the chip. As shown in FIG. 1, a third capture substance 6 for capturing the first capture substance is immobilized on the reference detection section 5 provided between the detection section 3 and the absorption pad 7. .
　第三の捕捉物質６は、第二の捕捉物質４と同様に、第一の捕捉物質とホスト－ゲストの関係にある物質（例えば、抗原、抗体、酵素、基質、リガンド、レセプター、ＤＮＡ、糖、ペプチドまたは合成高分子（例えばモレキュラーインプリントポリマー）など）であればよく、特に、抗体または合成高分子は、活性が安定しているので好ましい。第三の捕捉物質６を固定化する方法もまた、物理的吸着法、化学結合法または共有結合法などの公知の方法が適宜採用され得る。また固定化するためにスポッターを用いることが好ましい。特にラインプロットを行うことでライン状に抗体溶液を定量的に滴下することが可能となる。固定化後、風乾および熱風乾燥などにより、多孔体１を乾燥させる。多孔体１に第二の捕捉物質および第三の捕捉物質を固定化後、多孔体１全体を非特異吸着防止剤に浸すことで非特異吸着を抑えることが可能となる。非特異吸着防止剤として、例えばウシ血清アルブミン（ＢＳＡ）、カゼイン、ゼラチン、電気的に中性なポリマーやホスホリルコリン基を有するポリマーなど市販品の非特異吸着防止処理剤を用いることができる。ブロッキング処理後、自然乾燥、風乾、熱風乾燥または減圧乾燥などにより十分な乾燥を行う。 Similar to the second capture substance 4, the third capture substance 6 is a substance having a host-guest relationship with the first capture substance (eg, antigen, antibody, enzyme, substrate, ligand, receptor, DNA, sugar , A peptide or a synthetic polymer (for example, a molecular imprint polymer), and the like. In particular, an antibody or a synthetic polymer is preferable because its activity is stable. As a method for immobilizing the third capture substance 6, a known method such as a physical adsorption method, a chemical bonding method, or a covalent bonding method can be appropriately employed. Moreover, it is preferable to use a spotter for immobilization. In particular, by performing a line plot, it is possible to quantitatively drop the antibody solution in a line shape. After immobilization, the porous body 1 is dried by air drying or hot air drying. After immobilizing the second capture substance and the third capture substance on the porous body 1, nonspecific adsorption can be suppressed by immersing the entire porous body 1 in the nonspecific adsorption inhibitor. As the nonspecific adsorption inhibitor, for example, commercially available nonspecific adsorption inhibitor such as bovine serum albumin (BSA), casein, gelatin, an electrically neutral polymer or a polymer having a phosphorylcholine group can be used. After the blocking treatment, sufficient drying is performed by natural drying, air drying, hot air drying or reduced pressure drying.
　参照検出部５の構成は検出部３と同様であって、特に限定されず、対象物質の検出方法によって適宜決定され得る。例えば色の濃淡または発光（蛍光を含む）などに基づいて、対象物質を光学的に検出する場合、外部から色を観察できるような構造とすればよい。例えば、イムノクロマト用検査器具をハウジングした場合でも、観察窓を設けることで検出が可能となる。 The configuration of the reference detection unit 5 is the same as that of the detection unit 3 and is not particularly limited, and can be determined as appropriate depending on the detection method of the target substance. For example, when the target substance is optically detected based on color shading or light emission (including fluorescence), the structure may be such that the color can be observed from the outside. For example, even when an immunochromatographic inspection instrument is housed, detection is possible by providing an observation window.
　また、対象物質を電気化学的に検出する場合、参照検出部５は、多孔体１の上部または下部に形成された検出電極を有する検出手段を備えていればよい。検出電極は、少なくとも参照電極および作用電極の２電極から構成されていればよいが、参照電極および作用電極に加えて対向電極を備えている３電極から構成されていることが好ましい。 In addition, when the target substance is detected electrochemically, the reference detection unit 5 only needs to include detection means having detection electrodes formed on the upper or lower portion of the porous body 1. The detection electrode only needs to be composed of at least two electrodes, that is, a reference electrode and a working electrode, but is preferably composed of three electrodes provided with a counter electrode in addition to the reference electrode and the working electrode.
　また、参照検出部５の構成は、検出部３と同じ検出手段および／または検出構造を用いることが好ましい。同一の検出手段および／または検出構造であれば検出のバラつきの影響を無視することができる。また、検出信号値も温度により変動するが、検出手段および／または検出構造が同一であれば、変動幅も同一とみなすことが可能となる。 In addition, it is preferable that the configuration of the reference detection unit 5 uses the same detection means and / or detection structure as the detection unit 3. If the detection means and / or the detection structure are the same, the influence of detection variation can be ignored. The detection signal value also varies depending on the temperature. However, if the detection means and / or the detection structure are the same, the fluctuation range can be regarded as the same.
　＜１．５　標識された第一の捕捉物質＞
　流体中の物質を検出するための標識された第一の捕捉物質１０の標識物質は、ラテックス粒子やコロイド金属、色素、蛍光色素（または蛍光たんぱく質）、酵素または放射性物質など、ラベル化剤として使用できる物質であればよい。上記標識物質は微粒子であってもよい。上記構成によれば、色の濃淡等に基づいて微粒子の量を検出することができる。本明細書における「微粒子」の粒径は５０μｍ以下であれば特に限定されるものではないが、好ましくは０．０１～２μｍである。また、上記標識物質は電気化学的活性物質であってもよい。上記構成によれば、検出電極によって電気化学的に検出することができる。電気化学的活性物質の例としてはフェリシアン化物、フェロシアン化物、フェロセンおよびフェロセン誘導体などが挙げられる。 <1.5 Labeled first capture substance>
The labeling substance of the labeled first capturing substance 10 for detecting the substance in the fluid is used as a labeling agent such as latex particles, colloidal metals, dyes, fluorescent dyes (or fluorescent proteins), enzymes or radioactive substances. Any substance can be used. The labeling substance may be fine particles. According to the above configuration, the amount of fine particles can be detected based on color shading. The particle size of the “fine particles” in the present specification is not particularly limited as long as it is 50 μm or less, but is preferably 0.01 to 2 μm. The labeling substance may be an electrochemically active substance. According to the said structure, it can detect electrochemically with a detection electrode. Examples of the electrochemically active substance include ferricyanide, ferrocyanide, ferrocene and ferrocene derivatives.
　捕捉物質としては上記のように対象物質とホスト－ゲストの関係がある物質（例えば、抗原、抗体、酵素、基質、リガンド、レセプター、ＤＮＡ、糖、ペプチドまたは合成高分子（例えばモレキュラーインプリントポリマー）など）であればよく、特に、抗体または合成高分子は、活性が安定しているので好ましい。 As a capture substance, a substance having a host-guest relationship as described above (for example, antigen, antibody, enzyme, substrate, ligand, receptor, DNA, sugar, peptide, or synthetic polymer (for example, molecular imprint polymer) In particular, an antibody or a synthetic polymer is preferable because its activity is stable.
　＜１．５．１　コンジュゲーションパッド＞
　標識された第一の捕捉物質はコンジュゲーションパッド８により一時的に保存される。毛細管現象によりサンプルパッド２から移動してきた検体により一時的に保存されていた第一の捕捉物質が溶解しながら、検体に含まれる検出すべき物質を捕捉する部位となる。コンジュゲーションパッド８の材質としては、サンプルパッド２と同様に再生セルロース、酢酸セルロース、ニトロセルロース、ポリアクリロニトリル、エチレン酢酸ビニル、ポリウレタン、ポリメチルメタクリレート、ナイロン樹脂、ガラス繊維、パルプ、綿、レーヨン、アクリル、ポリエステルなどが例示される。 <1.5.1 Conjugation pad>
The labeled first capture substance is temporarily stored by the conjugation pad 8. The first capture substance temporarily stored by the specimen that has moved from the sample pad 2 due to capillary action is dissolved, and becomes a site for capturing the substance to be detected contained in the specimen. As the material of the conjugation pad 8, regenerated cellulose, cellulose acetate, nitrocellulose, polyacrylonitrile, ethylene vinyl acetate, polyurethane, polymethyl methacrylate, nylon resin, glass fiber, pulp, cotton, rayon, acrylic, as with the sample pad 2 And polyester.
　＜１．５．２　標識された第一の捕捉物質の量＞
　標識された第一の捕捉物質１０の保存量は、検出部３で第一の捕捉物質が十分に反応する量であり、かつ、参照検出部５でほぼ飽和で反応できる量とする。 <1.5.2 Amount of labeled first capture substance>
The storage amount of the labeled first capture substance 10 is an amount that allows the first capture substance to react sufficiently in the detection unit 3 and an amount that can be reacted in a substantially saturated manner in the reference detection unit 5.
　具体的には、第二の捕捉物質４および第三の捕捉物質６の固定化量がデータとして得られている場合、第一の捕捉物質の量は、下記の式で表すことが可能である。 Specifically, when the immobilized amounts of the second capture substance 4 and the third capture substance 6 are obtained as data, the amount of the first capture substance can be expressed by the following equation. .
　（第一の捕捉物質の保存量）≧（第二の捕捉物質４の固定化量）＋（第三の捕捉物質６の固定化量）
かつ（標識物質の保存量）≧（第二の捕捉物質４の固定化量）＋（第三の捕捉物質６の固定化量）
　ただし、第一の捕捉物質としてポリクローナル抗体など１つの対象物質に対し多数のエピトープで捕捉する捕捉物質を用いる場合、上記式の右辺にそれぞれ３倍した量を保存することが望ましい。 (Storage amount of the first capture substance) ≧ (immobilization amount of the second capture substance 4) + (immobilization amount of the third capture substance 6)
And (preservation amount of labeled substance) ≧ (immobilization amount of second capture substance 4) + (immobilization amount of third capture substance 6)
However, when a capture substance that captures a large number of epitopes for one target substance, such as a polyclonal antibody, is used as the first capture substance, it is desirable to store three times the amount on the right side of the above formula.
　検出部３における第二の捕捉物質４および参照検出部５における第三の捕捉物質６の固定化量のデータが得られていない場合、第一の捕捉物質は下記のように調製可能である。多孔体１自身の厚さまたは密度により第二の捕捉物質４および第三の捕捉物質６の固定化量は前後するため、基本的にこの方法で条件決定することが好ましい。具体的な調製例を下記に示す。 When data on the amount of immobilized second capture substance 4 in the detection unit 3 and third capture substance 6 in the reference detection unit 5 is not obtained, the first capture substance can be prepared as follows. Since the amount of immobilization of the second capture material 4 and the third capture material 6 varies depending on the thickness or density of the porous body 1 itself, it is basically preferable to determine the conditions by this method. Specific preparation examples are shown below.
　（１）参照検出部５の確認
　濃度既知の第一の捕捉物質（標準物質）を用いて、参照検出部５における第一の捕捉物質の検出範囲を調べる。得られた飽和濃度と送液した容量との積を導出することで、参照検出部５において第一の捕捉物質が飽和吸着するために必要な量を算出できる。 (1) Confirmation of Reference Detection Unit 5 Using the first capture substance (standard substance) whose concentration is known, the detection range of the first capture substance in the reference detection unit 5 is examined. By deriving the product of the obtained saturated concentration and the delivered volume, the reference detector 5 can calculate the amount necessary for the first trapping substance to be saturated and adsorbed.
　（２）検出部３の確認
　濃度既知の対象物質（標準物質）を用いて、検出部３において必要な第一の捕捉物質の最大量を調べる。具体的には想定される対象物質の最大濃度で反応させた検出部３において、検出部３上に捕捉された対象物質に対し飽和吸着するために必要な第一の捕捉物質濃度を調べる。得られた飽和濃度と容量との積を導出することで、第一の捕捉物質が飽和吸着するために必要な量を算出できる。 (2) Confirmation of the detection unit 3 Using the target substance (standard material) whose concentration is known, the maximum amount of the first capture substance necessary in the detection unit 3 is examined. Specifically, in the detection unit 3 that is reacted at the assumed maximum concentration of the target substance, the first trapping substance concentration necessary for saturated adsorption with respect to the target substance trapped on the detection unit 3 is examined. By deriving the product of the obtained saturated concentration and capacity, the amount necessary for the first trapping substance to be saturated and adsorbed can be calculated.
　以上、（１）、（２）から得られた第一の捕捉物質の量の和以上をコンジュゲーションパッド８に保存する。 As described above, the sum or more of the amount of the first capture substance obtained from (1) and (2) is stored in the conjugation pad 8.
　＜１．６　分析方法＞
　本実施形態に係るイムノクロマト用検査器具を用いた分析方法の例を、以下に示す。 <1.6 Analysis method>
An example of an analysis method using the immunochromatographic test instrument according to this embodiment is shown below.
　（１）サンプルの導入と反応
　図１に記載のイムノクロマト用検査器具に関して分析例を説明する。サンプルをサンプルパッド２に滴下する。サンプル中の対象物質は、毛管力によりサンプルパッド２からコンジュゲーションパッド８に移動する。コンジュゲーションパッド８に液体が流れ込むことで、標識された第一の捕捉物質が溶け出し、検体中に含まれる検出すべき物質を捕捉しながら、毛管力により多孔体１に流れ込む。第一の捕捉物質１０に捕捉された検出すべき物質（第一の捕捉物質１０と会合した検出すべき物質）は第二の捕捉物質４に捕捉される。一方で第二の捕捉物質４に捕捉されなかった第一の捕捉物質１０（検出すべき物質と会合していない第一の捕捉物質１０）は第三の捕捉物質６により捕捉される。 (1) Sample Introduction and Reaction An analysis example of the immunochromatographic test instrument shown in FIG. 1 will be described. A sample is dropped on the sample pad 2. The target substance in the sample moves from the sample pad 2 to the conjugation pad 8 by capillary force. When the liquid flows into the conjugation pad 8, the labeled first capture substance dissolves and flows into the porous body 1 by capillary force while capturing the substance to be detected contained in the specimen. The substance to be detected captured by the first capture substance 10 (the substance to be detected associated with the first capture substance 10) is captured by the second capture substance 4. On the other hand, the first capture substance 10 (first capture substance 10 not associated with the substance to be detected) not captured by the second capture substance 4 is captured by the third capture substance 6.
　現実的には、例えば第三の捕捉物質６の認識部位が第一の捕捉物質１０の認識部位と異なる部位を認識する場合、検出すべき物質と会合した第一の捕捉物質１０が第三の捕捉物質６に捕捉される可能性がある。しかし、このような場合でも、本願の作用効果は損なわれるものではない。 Actually, for example, when the recognition site of the third capture substance 6 recognizes a site different from the recognition site of the first capture substance 10, the first capture substance 10 associated with the substance to be detected is the third capture substance 10. There is a possibility of being trapped by the trapping substance 6. However, even in such a case, the effects of the present application are not impaired.
　（２）対象物質の検出
　検出は、検出部３および参照検出部５で行う。検出部３では検出部３上で捕捉された第一の捕捉物質１０の量（捕捉された対象物の量に相応）を検出し、参照検出部５では参照検出部５上で捕捉された第一の捕捉物質１０の量を検出する。 (2) Detection of target substance Detection is performed by the detection unit 3 and the reference detection unit 5. The detection unit 3 detects the amount of the first capture substance 10 captured on the detection unit 3 (corresponding to the amount of the captured object), and the reference detection unit 5 detects the first captured substance 10 captured on the reference detection unit 5. The amount of one capture substance 10 is detected.
　＜１．７　信号値の補正＞
　従来のイムノクロマト用検査器具の場合、検出部３の濃淡を目視にて確認し、定性的な測定を行ってきた。また、濃淡を光学的に読み取る装置を用いて定量測定が試みられている。しかしながら、正確な定量的検出には至っていない。その理由として、イムノクロマト用検査器具の場合、多孔体１の厚さ、密度の項が定量測定できない一番の要因となっている。 <1.7 Correction of signal value>
In the case of a conventional immunochromatographic inspection instrument, the light and shade of the detection unit 3 is visually confirmed, and qualitative measurement has been performed. In addition, quantitative measurement has been attempted using an apparatus that optically reads light and shade. However, accurate quantitative detection has not been achieved. The reason for this is that in the case of an immunochromatographic test instrument, the thickness and density terms of the porous body 1 are the primary factors that prevent quantitative measurement.
　例えば標識物質がラテックス粒子や金属コロイドの場合、検出信号値は下記のように表わされる。 For example, when the labeling substance is latex particles or metal colloids, the detection signal value is expressed as follows.
　・検出信号∝捕捉物質（第二の捕捉物質）の固定化密度（多孔体１密度に依存）×多孔体１の厚さ×抗原抗体反応率・・・（式１）
　また、標識物質が酵素の場合、検出信号値は下記のように表わされる。 Detection signal ∝ Immobilization density of capture substance (second capture substance) (depends on density of porous body 1) x thickness of porous body 1 x antigen-antibody reaction rate (Equation 1)
When the labeling substance is an enzyme, the detection signal value is expressed as follows.
　・検出信号∝（式１）×酵素基質反応率・・・（式２）
　下記にその他、信号値を変化させる要因の例を記す。 ・ Detection signal ∝ (formula 1) x enzyme substrate reaction rate (formula 2)
The following are examples of other factors that change the signal value.
Figure JPOXMLDOC01-appb-T000001
Figure JPOXMLDOC01-appb-T000001
　検出部３の信号値は、上記要因の他に、検体中に含まれる対象物質の濃度によって信号値が上下するが、参照検出部５の信号値は、必ず飽和吸着に相当する信号値が検出されるため、上記要因の影響のみを考慮することが可能となる。具体的には参照検出部５の信号値は下記のように表され得る。 In addition to the above factors, the signal value of the detection unit 3 varies depending on the concentration of the target substance contained in the sample, but the signal value of the reference detection unit 5 is always detected by a signal value corresponding to saturated adsorption. Therefore, only the influence of the above factors can be considered. Specifically, the signal value of the reference detection unit 5 can be expressed as follows.
　・参照検出部の信号値∝捕捉物質（第三の捕捉物質）の固定化密度（多孔体１密度に依存）×多孔体１の厚さ×抗原抗体反応率・・・（式３）
　上記の式１と比較すると使用環境に影響を受ける因子に関して相殺できるため、検出部３の信号値と参照検出部５の信号値とを比較することで、実際の値との相違を小さくすることが可能となる。補正の方法は、比をとることで可能であるが、その他、差や補正係数をかけるなど補正手段は問わない。 ・ Signal value of the reference detection part ∝ Immobilization density of the capture substance (third capture substance) (depending on the density of the porous body 1) × Thickness of the porous body 1 × antigen-antibody reaction rate (Equation 3)
Compared with Equation 1 above, factors that are affected by the use environment can be canceled out, so the difference between the actual value can be reduced by comparing the signal value of the detection unit 3 and the signal value of the reference detection unit 5. Is possible. The correction method can be performed by taking a ratio, but any other correction means such as applying a difference or a correction coefficient may be used.
　例えば、多孔体１の影響により２割減少していた場合、検出部３の信号値は最大２割減少することが考えられるが、参照検出部５の信号値も同じく減少するため、信号値で比較した場合、多孔体１自身の影響は無視できる。 For example, when it is reduced by 20% due to the influence of the porous body 1, the signal value of the detection unit 3 may be reduced by a maximum of 20%, but the signal value of the reference detection unit 5 also decreases, When compared, the influence of the porous body 1 itself can be ignored.
　また、補正後の検量線に関しては、器具のロットにより信号値が変化する可能性がある。そのため、ロット変更に伴い、ユーザーが、イムノクロマト用検査器具を検出するための装置またはソフトウェアに予め補正情報を入力しておくことで対応可能となる。また、チップにＩＣタグやバーコードなどを付加することで情報をチップに埋設しておき、検出装置側で当該情報を読みとり、自動で補正を行うことが可能である。例えば、検量線やデータ情報などをあらかじめ装置側に入力しておくことで、検出結果として得られた検出信号値から検量することが可能となるが、製造時のロットの違いや試薬のロットの違いにより検量線が異なる可能性があるため、検量するための情報を手入力またはバーコードやＩＣタグに入力しておくことができる。 Also, regarding the calibration curve after correction, the signal value may change depending on the lot of the instrument. Therefore, it becomes possible for the user to respond by inputting correction information in advance to an apparatus or software for detecting an immunochromatographic test instrument in accordance with the lot change. In addition, it is possible to embed information in the chip by adding an IC tag, a barcode, or the like to the chip, read the information on the detection device side, and automatically correct the information. For example, it is possible to perform calibration from the detection signal value obtained as a detection result by inputting a calibration curve, data information, etc. in advance on the device side. Since there is a possibility that the calibration curve differs depending on the difference, information for calibration can be input manually or in a bar code or IC tag.
　＜１．８　検出エラー＞
　参照検出部５は、イムノクロマト用検査器具での検出が失敗していないか否か判定することにも使用可能である。参照検出部５では、標識された第一の捕捉物質が飽和吸着したときの信号値が検出されるが、当該信号値が所定の値以下で検出された際に、測定自身の信頼性が低いと判断することが可能である。当該所定の値を基準値とすることができる。 <1.8 Detection error>
The reference detection unit 5 can also be used to determine whether or not the detection by the immunochromatographic test instrument has failed. The reference detection unit 5 detects a signal value when the labeled first capture substance is saturated and adsorbed, but when the signal value is detected below a predetermined value, the reliability of the measurement itself is low. It is possible to judge. The predetermined value can be used as a reference value.
　＜１．９　判定部＞
　図２に示すように、多孔体１上に第一の捕捉物質を捕捉するための第四の捕捉物質２１（図示せず）が固定化された判定部２０を設けてもよい。好ましくは参照検出部５よりも下流側に判定部２０を設ける。従来のイムノクロマト用検査器具のように、判定部２０の機能としては反応自身が完了したか否か判定することができる。また、判定部２０を設けることで、第一の捕捉物質の量の決定が容易になる。具体的には、判定部２０が標識物質により検出信号として検出されるだけの反応が必要となるため、１．５．２に記載したような調製方法をとらずとも、判定部２０の検出信号値が十分取れる条件で第一の捕捉物質の量を制御することが可能である。換言すれば、検出部３および参照検出部５で捕捉されなかった過剰な第一の捕捉物質が判定部２０まで流れてくるような量で、第一の捕捉物質の保存量を調整すればよい。 <1.9 Determination Unit>
As shown in FIG. 2, a determination unit 20 in which a fourth trapping substance 21 (not shown) for trapping the first trapping substance is immobilized on the porous body 1 may be provided. Preferably, the determination unit 20 is provided on the downstream side of the reference detection unit 5. As in the conventional immunochromatographic test instrument, as a function of the determination unit 20, it can be determined whether or not the reaction itself has been completed. In addition, by providing the determination unit 20, the amount of the first capture substance can be easily determined. Specifically, since the determination unit 20 needs to have a reaction that is detected as a detection signal by the labeling substance, the detection signal of the determination unit 20 can be obtained without taking the preparation method described in 1.5.2. It is possible to control the amount of the first capture substance under conditions that allow a sufficient value. In other words, the storage amount of the first capture substance may be adjusted by an amount such that excess first capture substance that has not been captured by the detection unit 3 and the reference detection unit 5 flows to the determination unit 20. .
　第四の捕捉物質としては、第一の捕捉物質とホスト－ゲストの関係がある物質（例えば、抗原、抗体、酵素、基質、リガンド、レセプター、ＤＮＡ、糖、ペプチドまたは合成高分子（例えばモレキュラーインプリントポリマー）など）であればよく、特に、抗体または合成高分子は、活性が安定しているので好ましい。また、第四の捕捉物質は第三の捕捉物質と同一の捕捉物質であることが好ましい。上記構成によれば、判定部を参照検出部と同様の構成で形成することができ、イムノクロマト用検査器具の構成を簡易化することができる。 The fourth capture substance includes a substance having a host-guest relationship with the first capture substance (eg, antigen, antibody, enzyme, substrate, ligand, receptor, DNA, sugar, peptide, or synthetic polymer (eg, molecular in Print polymer) and the like. In particular, an antibody or a synthetic polymer is preferable because its activity is stable. Further, the fourth capture substance is preferably the same capture substance as the third capture substance. According to the said structure, a determination part can be formed with the structure similar to a reference detection part, and the structure of the test | inspection instrument for immunochromatography can be simplified.
　〔実施の形態２〕
　図３に基づいて、本発明の実施の形態２について説明する。なお、実施の形態１と同じ構成については、ここでは、その詳細な説明を省略する。 [Embodiment 2]
A second embodiment of the present invention will be described with reference to FIG. In addition, about the same structure as Embodiment 1, the detailed description is abbreviate | omitted here.
　実施の形態１においては、参照検出部５と検出部３とは、同じ多孔体１に直列に配置されるが、実施の形態２においては、図３のように、参照検出部５’と検出部３’とは、分岐した多孔体１に並列に配置される。並列に配置することで、「標識された第一の捕捉物質の保存量」の項目が実施の形態１とは異なるが、その他の項目に関しては実施の形態１に従う。 In the first embodiment, the reference detection unit 5 and the detection unit 3 are arranged in series in the same porous body 1, but in the second embodiment, as shown in FIG. 3, the reference detection unit 5 ′ and the detection unit 3 are detected. The part 3 ′ is arranged in parallel with the branched porous body 1. By arranging them in parallel, the item “preserved amount of labeled first capture substance” is different from that in the first embodiment, but other items are the same as those in the first embodiment.
　多孔体１が分岐する数としては特に限定されず、適宜設定され得る。例えば、２本であってもよく、３本であってもよく、４本であってもよく、それよりも多くても良い。分岐する数を２本よりも多くする場合には、例えば、数を偶数個（例えば、４、６、８、・・・）にすることが可能である。なお、この場合、各検出部３’と各参照検出部５’とが対をなすように、同じ数の検出部３’と参照検出部５’とを設けることが好ましい。上記構成によれば、１つのイムノクロマト用検査器具によって、複数種類の物質を同時に検出および分析することが可能になる。この場合、各検出部３’および参照検出部５’は、検出および分析したい物質に合わせた構成にすることが可能である。つまり、複数の検出部３’は、各々が同じ構成を有する検出部３’であっても良いが、各々が異なる構成を有する検出部３’であっても良い。同様に、複数の参照検出部５’は、各々が同じ構成を有する参照検出部５’であっても良いが、各々が異なる構成を有する参照検出部５’であっても良い。 The number of branches of the porous body 1 is not particularly limited and can be set as appropriate. For example, the number may be two, three, four, or more. When the number of branches is more than two, for example, the number can be an even number (for example, 4, 6, 8,...). In this case, it is preferable to provide the same number of detection units 3 ′ and reference detection units 5 ′ so that each detection unit 3 ′ and each reference detection unit 5 ′ make a pair. According to the above configuration, it is possible to simultaneously detect and analyze a plurality of types of substances with one immunochromatographic test instrument. In this case, each detection unit 3 ′ and reference detection unit 5 ′ can be configured according to the substance to be detected and analyzed. That is, the plurality of detection units 3 ′ may be detection units 3 ′ having the same configuration, but may be detection units 3 ′ having different configurations. Similarly, the plurality of reference detection units 5 ′ may be reference detection units 5 ′ having the same configuration, but may be reference detection units 5 ′ having different configurations.
　分岐した各々の多孔体の、流体が移動する方向に対して垂直な方向における断面の大きさおよび形状は特に限定されないが、同じ大きさおよび形状であることが好ましい。上記構成によれば、精度よく物質を検出および分析することが可能である。 The size and shape of the cross section of each branched porous body in the direction perpendicular to the direction in which the fluid moves are not particularly limited, but are preferably the same size and shape. According to the above configuration, it is possible to detect and analyze a substance with high accuracy.
　＜２．１　標識された第一の捕捉物質の量＞
　標識された第一の捕捉物質の保存量は、実施の形態１と同様に、検出部３’で第一の捕捉物質が十分反応する量であり、かつ、参照検出部５’でほぼ飽和で反応できる量とする。 <2.1 Amount of labeled first capture substance>
As in the first embodiment, the storage amount of the labeled first capture substance is an amount that allows the first capture substance to react sufficiently in the detection unit 3 ′, and is almost saturated in the reference detection unit 5 ′. The amount can be reacted.
　具体的には、検出部３’における第二の捕捉物質４’（図示せず）および参照検出部５’における第三の捕捉物質６’（図示せず）の固定化量がデータとして得られている場合、第一の捕捉物質の量は下記の式で表すことが可能である。 Specifically, the immobilized amounts of the second capture substance 4 ′ (not shown) in the detection unit 3 ′ and the third capture substance 6 ′ (not shown) in the reference detection unit 5 ′ are obtained as data. The amount of the first capture substance can be represented by the following formula:
　（第二の捕捉物質４’の固定化量）≧（第三の捕捉物質６’の固定化量）の場合
　（第一の捕捉物質の保存量）≧（第二の捕捉物質４’の固定化量）×２
　（第三の捕捉物質６’の固定化量）≧（第二の捕捉物質４’の固定化量）の場合
　（第一の捕捉物質の保存量）≧（第三の捕捉物質６’の固定化量）×２
　図３に示された構成は２分岐流路であるためチップ全体として２倍保存する。ｎ分岐の場合はｎ倍行う。 (Immobilization amount of second capture substance 4 ′) ≧ (immobilization amount of third capture substance 6 ′) (preservation quantity of first capture substance) ≧ (immobilization of second capture substance 4 ′) Amount) x 2
(Immobilization amount of third capture substance 6 ′) ≧ (immobilization amount of second capture substance 4 ′) (preservation amount of first capture substance) ≧ (fixation of third capture substance 6 ′) Amount) x 2
Since the configuration shown in FIG. 3 is a two-branch channel, the entire chip is stored twice. In the case of n branches, it is performed n times.
　ただし、第一の捕捉物質として、ポリクローナル抗体など１つの対象物質に対し多数のエピトープで捕捉する捕捉物質を用いる場合、上記式の３倍程度第一の捕捉物質を保存することが望ましい。 However, when a capture substance that captures a large number of epitopes for one target substance, such as a polyclonal antibody, is used as the first capture substance, it is desirable to store the first capture substance about three times the above formula.
　検出部３’における第二の捕捉物質４’および参照検出部５’における第三の捕捉物質６’の固定化量データが得られていない場合、第一の捕捉物質は下記のように調製可能である。具体的な調製例を下記に示す。
（１）参照検出部５’の確認
　濃度既知の第一の捕捉物質（標準物質）を用いて参照検出部５’における第一の捕捉物質の検出範囲を調べる。得られた飽和濃度から容量との積を導出することで、参照検出部５’において第一の捕捉物質が飽和吸着するために必要な量を算出できる。
（２）検出部３’の確認
　濃度既知の対象物質（標準物質）を用いて検出部３’において必要な第一の捕捉物質の最大量を調べる。具体的には想定される対象物質の最大濃度で反応させた検出部３’において、検出部３’上に捕捉された対象物質に対し飽和吸着するために必要な第一の捕捉物質濃度を調べる。得られた飽和濃度と容量との積を導出することで、第一の捕捉物質が飽和吸着するために必要な量を算出できる。 When the immobilized amount data of the second capture substance 4 ′ in the detection unit 3 ′ and the third capture substance 6 ′ in the reference detection unit 5 ′ are not obtained, the first capture substance can be prepared as follows. It is. Specific preparation examples are shown below.
(1) Confirmation of reference detection unit 5 ′ Using the first capture substance (standard substance) having a known concentration, the detection range of the first capture substance in the reference detection unit 5 ′ is examined. By deriving the product of the capacity from the obtained saturated concentration, the amount necessary for the first capture substance to be saturated and adsorbed in the reference detection unit 5 ′ can be calculated.
(2) Confirmation of detection part 3 'The maximum amount of the first capture substance required in the detection part 3' is examined using a target substance (standard substance) whose concentration is known. Specifically, in the detection unit 3 ′ reacted at the maximum concentration of the target substance assumed, the concentration of the first capture substance necessary for saturated adsorption to the target substance captured on the detection unit 3 ′ is examined. . By deriving the product of the obtained saturated concentration and capacity, the amount necessary for the first trapping substance to be saturated and adsorbed can be calculated.
　以上、（１）、（２）から得られた第一の捕捉物質の量を比較し、多かった方の２倍量保存する。
〔実施の形態３〕
　＜３．分析装置＞
　本発明に係る分析装置は、本発明に係るイムノクロマト用検査器具を用いた分析方法を実現できる装置であれば、その構成はどのようなものでも構わないが、イムノクロマト用検査器具における標識物質が発する信号を検出する検出器（検出手段）及び演算部を備えている。また、本発明に係る分析装置は、本発明に係るイムノクロマト用検査器具を備えたものであってもよい。 As described above, the amount of the first capture substance obtained from (1) and (2) is compared, and the amount twice as large as the larger amount is stored.
[Embodiment 3]
<3. Analyzer>
The analyzer according to the present invention may have any configuration as long as it can realize the analysis method using the immunochromatographic test instrument according to the present invention, but the labeling substance in the immunochromatographic test instrument emits. A detector (detection means) for detecting a signal and a calculation unit are provided. Moreover, the analyzer according to the present invention may be provided with the immunochromatographic test instrument according to the present invention.
　（１）検出器（検出手段）
　本発明に係る分析装置は、本発明に係るイムノクロマト用検査器具の検出部３の信号値および参照検出部５の信号値を検出する検出器を備えている。検出器の構成は特に限定されず、検出部３および参照検出部５と同様に、対象物質の検出方法によって適宜決定され得る。 (1) Detector (detection means)
The analyzer according to the present invention includes a detector that detects the signal value of the detection unit 3 and the signal value of the reference detection unit 5 of the immunochromatographic test instrument according to the present invention. The configuration of the detector is not particularly limited, and can be appropriately determined according to the detection method of the target substance, similarly to the detection unit 3 and the reference detection unit 5.
　また、検出部３に対応する検出器および参照検出部５に対応する検出器における検出手段は同じであることが好ましい。上記構成であれば、同一の検出手段を用いるため、共通の検出器を用いることができ、分析装置の構成を簡易化することができる。 Also, it is preferable that the detection means in the detector corresponding to the detector 3 and the detector corresponding to the reference detector 5 are the same. If it is the said structure, since the same detection means is used, a common detector can be used and the structure of an analyzer can be simplified.
　また、本発明に係る分析装置は、判定部からの信号を検出する検出器を備えていてもよい。判定部から信号が検出されない間は、反応が終わっていないと判定し、検出処理を継続する。判定部から所定の信号が検出されれば、反応が完了したと判定し、検出処理を終了し、演算部にて信号値の補正および検量値の算出を行う。 Further, the analysis apparatus according to the present invention may include a detector that detects a signal from the determination unit. While no signal is detected from the determination unit, it is determined that the reaction has not ended, and the detection process is continued. If a predetermined signal is detected from the determination unit, it is determined that the reaction is completed, the detection process is terminated, and the calculation unit corrects the signal value and calculates the calibration value.
　（２）演算部
　演算部は、＜１．７　信号値の補正＞に記載された方法に従って、検出部から検出した信号値と参照検出部から検出した信号値とを比較し、検出信号値（相対信号値）を算出する。その後、あらかじめ装置側に入力された検量線やデータ情報などに基づき、検出結果として得られた検出信号値から検量値（検出すべき物質の濃度）を算出することが可能となる。 (2) Calculation Unit The calculation unit compares the signal value detected from the detection unit with the signal value detected from the reference detection unit according to the method described in <1.7 Correction of Signal Value>, and detects the detection signal value ( Relative signal value) is calculated. Thereafter, a calibration value (concentration of a substance to be detected) can be calculated from a detection signal value obtained as a detection result based on a calibration curve, data information, or the like previously input to the apparatus side.
　本発明に係る分析装置は、上記の構成の他に下記の構成を備えていてもよい。 The analyzer according to the present invention may have the following configuration in addition to the above configuration.
　（３）表示部
　本発明に係る分析装置は、検量値を表示する表示部を備えていてもよい。当該表示部はディスプレイであってもよいし、演算結果をプリントアウトするものであってもよい。 (3) Display Unit The analyzer according to the present invention may include a display unit that displays a calibration value. The display unit may be a display, or may print out a calculation result.
　（４）警告発生部
　本発明に係る分析装置は警告発生部を備えていることが好ましい。＜１．８　検出エラー＞に記載したように分析装置での検出時に参照検出部５の信号値が基準値を下回った場合、ユーザーに警告を発するとともに、表示部にて検量値の表示を行わないようにすることで、ユーザーに誤った検量値を伝えることを防止することが可能になる。警告は光による警告でもよいし、音による警告であってもよく、従来周知の方法を用いることが可能である。ただし、検出部および参照検出部において、光による検出が行われる場合、それらに影響を及ぼさないように、音による警告が好ましい。 (4) Warning generator The analyzer according to the present invention preferably includes a warning generator. As described in <1.8 Detection error>, when the signal value of the reference detection unit 5 falls below the standard value during detection by the analyzer, a warning is given to the user and the calibration value is displayed on the display unit. By avoiding this, it is possible to prevent an erroneous calibration value from being transmitted to the user. The warning may be a light warning or a sound warning, and a conventionally known method can be used. However, when detection by light is performed in the detection unit and the reference detection unit, a warning by sound is preferable so as not to affect them.
　（５）バーコードまたはＩＣタグ用リーダー
　本発明に係る分析装置はバーコードまたはＩＣタグ用のリーダーを備えていることが好ましい。チップのＩＣタグやバーコードなどには、製造時のロットの違いや試薬のロットの違い等の検量するための情報が埋設されている。分析装置側では、バーコードやＩＣタグに入力された情報を演算部に反映させ、検量線またはデータの補正を行うことが可能となる。分析装置によるバーコードやＩＣタグの読み取り方法は、あらゆる周知の技術を用いることが可能である。 (5) Barcode or IC Tag Reader The analyzer according to the present invention preferably includes a barcode or IC tag reader. Embedded in the IC tag, barcode, etc. of the chip are information for calibration such as differences in lots at the time of manufacture and differences in lots of reagents. On the analyzer side, it is possible to correct the calibration curve or the data by reflecting the information input to the barcode or IC tag on the calculation unit. Any known technique can be used as a method of reading a barcode or IC tag by the analyzer.
　本発明は上述した各実施形態に限定されるものではなく、請求項に示した範囲で種々の変更が可能であり、異なる実施形態にそれぞれ開示された技術的手段を適宜組み合わせて得られる実施形態についても本発明の技術的範囲に含まれる。 The present invention is not limited to the above-described embodiments, and various modifications are possible within the scope shown in the claims, and embodiments obtained by appropriately combining technical means disclosed in different embodiments. Is also included in the technical scope of the present invention.
　本発明は、免疫学的クロマトグラフィー（イムノクロマト法）を用いて、検体中の測定すべき物質を定量的に検出するための器具、装置および方法に関する。 The present invention relates to an instrument, an apparatus and a method for quantitatively detecting a substance to be measured in a specimen using immunological chromatography (immunochromatography).
　１、１’　　　　　多孔体
　２、２’　　　　　サンプルパッド
　３、３’　　　　　検出部
　４　　　　　　　　第二の捕捉物質
　５、５’　　　　　参照検出部
　６　　　　　　　　第三の捕捉物質
　７、７’、７’’　吸収パッド
　８、８’　　　　　コンジュゲーションパッド
　９　　　　　　　　標識物質が修飾された第一の捕捉物質
　１０　　　　　　　第一の捕捉物質
　１５　　　　　　　バッキングシート
　２０　　　　　　　判定部
　１０１　　　　　　多孔体
　１０２　　　　　　サンプルパッド
　１０３　　　　　　検出部
　１０４　　　　　　第二の捕捉物質
　１０５　　　　　　検出すべき物質
　１０７　　　　　　吸収パッド
　１０８　　　　　　コンジュゲーションパッド
　１０９　　　　　　抗体感作ラテックス
　１１０　　　　　　第一の捕捉物質
　１１１　　　　　　抗原抗体複合体
　１１５　　　　　　バッキングシート DESCRIPTION OF SYMBOLS 1, 1 'Porous body 2, 2' Sample pad 3, 3 'Detection part 4 Second capture substance 5, 5' Reference detection part 6 Third capture substance 7, 7 ', 7''Absorption pad 8, 8 'Conjugation pad 9 First capture substance with modified labeling substance 10 First capture substance 15 Backing sheet 20 Determination part 101 Porous body 102 Sample pad 103 Detection part 104 Second capture substance 105 Substance to be detected Absorption Pad 108 conjugation pad 109 antibody-sensitized latex 110 first capture substance 111 antigen-antibody complex 115 backing sheet

Claims (28)

  1. 　検体が毛管現象により移動可能な多孔体を有するイムノクロマト用検査器具において、
    　検出すべき物質を捕捉するとともに標識物質が結合している第一の捕捉物質が移動可能なように配置された領域と、
    　上記検出すべき物質を特異的に捕捉する第二の捕捉物質が、上記多孔体上に固定された状態で配置された検出部と、
    　上記多孔体上に設けられているとともに、上記第一の捕捉物質を捕捉するための第三の捕捉物質が配置されている参照検出部と、が設けられており、
    　上記第三の捕捉物質は、飽和吸着にて上記第一の捕捉物質を捕捉することを特徴とするイムノクロマト用検査器具。     In an immunochromatographic test instrument having a porous body in which a specimen can move by capillary action,
    An area where the first capture substance to which the substance to be detected is captured and the labeling substance is bound is arranged to be movable;
    A second capturing substance that specifically captures the substance to be detected, and a detection unit arranged in a state of being fixed on the porous body;
    A reference detection unit provided on the porous body and provided with a third capture substance for capturing the first capture substance, and
    The immunochromatographic test instrument, wherein the third capture substance captures the first capture substance by saturated adsorption.
  2. 　上記参照検出部は、上記検出部の下流側に設けられていることを特徴とする請求項１に記載のイムノクロマト用検査器具。 The immunochromatographic test instrument according to claim 1, wherein the reference detection unit is provided downstream of the detection unit.
  3. 　上記多孔体に流される上記第一の捕捉物質の量および上記第一の捕捉物質に結合している標識物質の量は、それぞれ上記第二の捕捉物質の量と上記第三の捕捉物質の量との和以上の量であることを特徴とする請求項２に記載のイムノクロマト用検査器具。 The amount of the first capture substance and the amount of the labeling substance bound to the first capture substance flowing through the porous body are respectively the amount of the second capture substance and the amount of the third capture substance. The immunochromatographic test instrument according to claim 2, wherein the amount is greater than or equal to the sum.
  4. 　上記多孔体は、分岐しており、
    　上記分岐した多孔体の１つに、上記検出部が設けられており、
    　上記分岐した多孔体のうち、上記検出部が設けられた多孔体とは別の多孔体に、上記参照検出部が設けられていることを特徴とする請求項１に記載のイムノクロマト用検査器具。     The porous body is branched,
    One of the branched porous bodies is provided with the detection unit,
    2. The immunochromatographic test instrument according to claim 1, wherein, in the branched porous body, the reference detection section is provided in a porous body different from the porous body provided with the detection section.
  5. 　上記多孔体に流される上記第一の捕捉物質の量および上記第一の捕捉物質に結合している標識物質の量は、それぞれ上記検出部における上記第二の捕捉物質の量または上記参照検出部における上記第三の捕捉物質の量の２倍以上であることを特徴とする請求項４に記載のイムノクロマト用検査器具。 The amount of the first capture substance flowing into the porous body and the amount of the labeling substance bound to the first capture substance are the amount of the second capture substance in the detection unit or the reference detection unit, respectively. The immunochromatographic test instrument according to claim 4, wherein the test instrument is at least twice the amount of the third capture substance.
  6. 　上記参照検出部および上記検出部は、同一の検出手段を備えていることを特徴とする請求項１～５の何れか１項に記載のイムノクロマト用検査器具。 The immunochromatographic test instrument according to any one of claims 1 to 5, wherein the reference detection unit and the detection unit include the same detection means.
  7. 　上記参照検出部および上記検出部は、外部から色の濃淡または発光を観察できる構造であることを特徴とする請求項１～６の何れか１項に記載のイムノクロマト用検査器具。 The immunochromatographic test instrument according to any one of claims 1 to 6, wherein the reference detection unit and the detection unit have a structure capable of observing color shading or light emission from the outside.
  8. 　上記参照検出部および上記検出部は、それぞれ作用電極および参照電極を備えていることを特徴とする請求項１～６の何れか１項に記載のイムノクロマト用検査器具。 The immunochromatographic test instrument according to any one of claims 1 to 6, wherein each of the reference detection unit and the detection unit includes a working electrode and a reference electrode.
  9. 　上記作用電極および上記参照電極は、大きさが同一であることを特徴とする請求項８に記載のイムノクロマト用検査器具。 9. The immunochromatographic test instrument according to claim 8, wherein the working electrode and the reference electrode have the same size.
  10. 　上記第一の捕捉物質が、上記検出すべき物質に対する抗体であることを特徴とする請求項１～９の何れか１項に記載のイムノクロマト用検査器具。 10. The immunochromatographic test instrument according to claim 1, wherein the first capture substance is an antibody against the substance to be detected.
  11. 　上記第二の捕捉物質が、上記検出すべき物質に対する抗体であることを特徴とする請求項１～１０の何れか１項に記載のイムノクロマト用検査器具。 The immunochromatographic test instrument according to any one of claims 1 to 10, wherein the second capture substance is an antibody against the substance to be detected.
  12. 　上記第三の捕捉物質が、上記第一の捕捉物質に対する抗体であることを特徴とする請求項１～１１の何れか１項に記載のイムノクロマト用検査器具。 12. The immunochromatographic test instrument according to claim 1, wherein the third capture substance is an antibody against the first capture substance.
  13. 　上記標識物質が、微粒子であることを特徴とする請求項１～１２の何れか１項に記載のイムノクロマト用検査器具。 The immunochromatographic test instrument according to any one of claims 1 to 12, wherein the labeling substance is a fine particle.
  14. 　上記標識物質が、蛍光材料であることを特徴とする請求項１～１３の何れか１項に記載のイムノクロマト用検査器具。 14. The immunochromatographic test instrument according to claim 1, wherein the labeling substance is a fluorescent material.
  15. 　上記標識物質が、電気化学的活性物質であることを特徴とする請求項１～１４の何れか１項に記載のイムノクロマト用検査器具。 The immunochromatographic test instrument according to any one of claims 1 to 14, wherein the labeling substance is an electrochemically active substance.
  16. 　上記参照検出部の下流に、上記第一の捕捉物質を捕捉するための第四の捕捉物質が配置されている判定部が設けられていることを特徴とする請求項１～１５の何れか１項に記載のイムノクロマト用検査器具。 The determination unit according to any one of claims 1 to 15, wherein a determination unit in which a fourth capture substance for capturing the first capture substance is disposed downstream of the reference detection unit. The immunochromatographic test device according to Item.
  17. 　上記検出部の信号値および上記参照検出部の信号値から導出される検出信号値と、上記検出すべき物質の濃度との相関関係に関する情報が書き込まれたバーコードまたはＩＣタグが設けられていることを特徴とする請求項１～１６の何れか１項に記載のイムノクロマト用検査器具。 A barcode or IC tag is provided in which information relating to the correlation between the detection signal value derived from the signal value of the detection unit and the signal value of the reference detection unit and the concentration of the substance to be detected is written. The immunochromatographic test instrument according to any one of claims 1 to 16, wherein
  18. 　請求項１～１７の何れか１項に記載のイムノクロマト用検査器具における上記標識物質が発する信号を検出する検出器を備えていることを特徴とする分析装置。 18. An analyzer comprising a detector for detecting a signal emitted from the labeling substance in the immunochromatographic test instrument according to any one of claims 1 to 17.
  19. 　上記検出部の信号値と上記参照検出部の信号値とを比較した相対信号値から、上記検出すべき物質の濃度を算出することを特徴とする請求項１８に記載の分析装置。 The analyzer according to claim 18, wherein the concentration of the substance to be detected is calculated from a relative signal value obtained by comparing the signal value of the detection unit and the signal value of the reference detection unit.
  20. 　上記参照検出部の信号値が基準値以下である場合に、警告を発することを特徴とする請求項１８または１９に記載の分析装置。 The analyzer according to claim 18 or 19, wherein a warning is issued when a signal value of the reference detection unit is equal to or less than a reference value.
  21. 　上記検出部の信号値および上記参照検出部の信号値について、検出する検出器は同じであることを特徴とする請求項１８～２０の何れか１項に記載の分析装置。 The analyzer according to any one of claims 18 to 20, wherein the detector to detect the signal value of the detection unit and the signal value of the reference detection unit are the same.
  22. 　バーコードまたはＩＣタグ用のリーダーが設けられていることを特徴とする請求項１８～２１の何れか１項に記載の分析装置。 The analyzer according to any one of claims 18 to 21, wherein a reader for barcode or IC tag is provided.
  23. 　請求項１～１７の何れか１項に記載のイムノクロマト用検査器具を用い、
    　上記参照検出部の信号値と上記検出部の信号値とを比較して、検出信号値を算出することを特徴とする分析方法。     Using the immunochromatographic test instrument according to any one of claims 1 to 17,
    An analysis method comprising: calculating a detection signal value by comparing a signal value of the reference detection unit with a signal value of the detection unit.
  24. 　上記検出信号値は、上記検出部の信号値と上記参照検出部の信号値との比から算出されることを特徴とする請求項２３に記載の分析方法。 24. The analysis method according to claim 23, wherein the detection signal value is calculated from a ratio between the signal value of the detection unit and the signal value of the reference detection unit.
  25. 　上記参照検出部の信号値が基準値以下である場合に、検出エラーと判断することを特徴とする請求項２３または２４に記載の分析方法。 25. The analysis method according to claim 23 or 24, wherein a detection error is determined when the signal value of the reference detection unit is equal to or less than a reference value.
  26. 　上記検出信号値と上記検出すべき物質の濃度との相関関係を予め上記イムノクロマト用検査器具へ入力し、
    　上記検出信号値と上記相関関係とから、上記検出すべき物質の濃度を算出することを特徴とする請求項２３～２５の何れか１項に記載の分析方法。     The correlation between the detection signal value and the concentration of the substance to be detected is input in advance to the immunochromatographic test instrument,
    The analysis method according to any one of claims 23 to 25, wherein the concentration of the substance to be detected is calculated from the detection signal value and the correlation.
  27. 　上記イムノクロマト用検査器具の製造ロットおよび試薬ロットの少なくとも一方に関する情報に基づいて、上記相関関係を補正することを特徴とする請求項２６に記載の分析方法。 27. The analysis method according to claim 26, wherein the correlation is corrected based on information relating to at least one of a production lot and a reagent lot of the immunochromatographic test instrument.
  28. 　上記情報を、上記イムノクロマト用検査器具に設けられたバーコードまたはＩＣタグから読み取ることを特徴とする請求項２７に記載の分析方法。 28. The analysis method according to claim 27, wherein the information is read from a barcode or an IC tag provided in the immunochromatographic test instrument.
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