WO2013122121A1 - Instrument d'essai pour immunochromatographie, analyseur et procédé d'analyse - Google Patents

Instrument d'essai pour immunochromatographie, analyseur et procédé d'analyse 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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English (en)
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

L'objectif de la présente invention est de détecter quantitativement un échantillon contenant un analyte à l'aide d'un instrument d'essai pour immunochromatographie. L'instrument d'essai pour immunochromatographie est pourvu d'une unité de détection de substance de référence qui permet de détecter exclusivement l'activité d'une substance marquée qui a été capturée. Étant donné que l'unité de détection de substance de référence détecte de manière certaine une valeur de signal correspondant à un volume de capture maximal, les effets provoqués par des variations d'épaisseur et de densité d'un matériau poreux peuvent être éliminés par comparaison de la valeur de signal susmentionnée avec une valeur de signal détectée par une unité de détection.
PCT/JP2013/053463 2012-02-17 2013-02-14 Instrument d'essai pour immunochromatographie, analyseur et procédé d'analyse WO2013122121A1 (fr)

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WO2019215199A1 (fr) 2018-05-07 2019-11-14 Immundiagnostik Ag Système d'analyse par immunochromatographie sur membrane quantitative
WO2020165456A1 (fr) 2019-02-15 2020-08-20 Immundiagnostik Ag Test rapide pour le diagnostic d'infections bactériennes chez les nouveau-nés
WO2022097627A1 (fr) * 2020-11-05 2022-05-12 国立大学法人浜松医科大学 Procédé de fabrication d'un kit de test immunochromatographique

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JP6183547B2 (ja) 2014-04-23 2017-08-23 日本精工株式会社 ダイレクトドライブモータ、搬送装置、検査装置、及び、工作機械
US11536707B2 (en) 2014-09-23 2022-12-27 Tearlab Research, Inc. Systems and methods for integration of microfluidic tear collection and lateral flow analysis of analytes of interest
WO2017003368A1 (fr) * 2015-06-29 2017-01-05 Cell Id Pte Ltd Procédé et appareil permettant de collecter des données de test d'utilisation d'un kit de test jetable
JP6429746B2 (ja) * 2015-07-31 2018-11-28 国立研究開発法人産業技術総合研究所 電気化学的分析用電極およびその電極を用いた電気化学的分析用キット
JP2019113425A (ja) * 2017-12-25 2019-07-11 Ｂｌｕｅ Ｉｎｄｕｓｔｒｉｅｓ株式会社 イムノクロマトアッセイ用検査カートリッジ及び検査装置

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JPH055743A (ja) * 1991-01-31 1993-01-14 Wakunaga Pharmaceut Co Ltd 測定装置

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2019215199A1 (fr) 2018-05-07 2019-11-14 Immundiagnostik Ag Système d'analyse par immunochromatographie sur membrane quantitative
WO2020165456A1 (fr) 2019-02-15 2020-08-20 Immundiagnostik Ag Test rapide pour le diagnostic d'infections bactériennes chez les nouveau-nés
WO2022097627A1 (fr) * 2020-11-05 2022-05-12 国立大学法人浜松医科大学 Procédé de fabrication d'un kit de test immunochromatographique

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