JP2002048798A - Measuring method by using labeled particle containing metal or metal compound - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a measuring method and a measuring device capable of measuring easily a measuring object. SOLUTION: The measuring object is coupled with a labeled particle containing a metal or a metal compound through an antibody or an antigen coupled with the labeled particle, and the metal quantity of the labeled particle coupled with the measuring object is measured by measuring an eddy current or a capacitance, to thereby measure the measuring object.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

The present invention relates to a method for measuring an object to be measured using a biologically specific reaction such as an antigen-antibody reaction. More specifically, the present invention relates to a method for measuring an object to be measured using labeled particles containing a metal or a metal compound, a test piece used for the measurement method, and a measuring device adapted to the test piece.

[0002]

2. Description of the Related Art As a measurement method using a biologically specific reaction, there is an immunoassay method using an antigen-antibody reaction.
There are various immunoassay methods using an antigen-antibody reaction, and radioactive isotopes as labeling substances for antibodies or antigens,
Radioimmunoassay (RI) using insoluble carrier particles such as enzymes, fluorescent substances, luminescent substances, latex, etc.
A), enzyme immunoassay (EIA), fluorescence immunoassay (FIA), luminescence immunoassay (LIA), latex agglutination immunoassay (LPIA) and the like are known.

As a measurement method using fine particles containing a metal or a metal compound, an operation of separating and washing a reaction compound and an unreacted substance using particles containing a magnetic compound (B)
/ F separation) is known. Specifically, after mixing the measurement sample and the magnetic compound-containing particles carrying an antibody or antigen capable of reacting with the measurement target contained therein, the magnetic compound-containing particles are attached to the reaction vessel wall by the action of a magnetic field, This is a method of removing and washing the reactants (J. Boland et al., Clinical Chemistry, 36 (9), 1598, 19).
90). In this method, the magnetic compound-containing particles are used only for attaching the measurement object to the reaction vessel wall, and do not detect the measurement object by the contained metal or metal compound. In the case of this method, a secondary reaction with a label using an enzyme, fluorescence, or the like is required for detecting the measurement object.

[0004] Immunochromatography is widely used as a measurement method suitable for simple inspection without using a dedicated device for measurement (Japanese Patent Application Laid-Open Nos. 1-3169, Hei.
-184840). That is, a nitrocellulose membrane, a glass filter membrane, a nylon membrane, a modified nylon membrane, a membrane material such as a polyvinylidene difluorolide (PVDF) membrane, a filter paper, a nonwoven fabric (for example, Using sheet-like materials such as cellulosic fibers, glass fibers, or those prepared from chemical fibers such as nylon, polyester, polypropylene, and polyurethane), and using colloidal gold or colored latex particles as markers, measurement The undiluted solution of the sample or its diluted solution is moved along with the label by capillary action on the membrane surface of the stationary phase, and the antibody or antigen immobilized on the stationary phase, the measurement object, and the antibody or antigen further immobilized on the label are Changes in the color of the reaction site utilizing the fact that the label remains at the reaction site on the stationary phase as a result of the reaction A method for the detection of the measuring object performed by determining by visual observation.

[0005]

However, in the prior art, there are various problems regarding the measuring method. For example, the RIA has a safety problem with respect to handling and disposal of radioisotopes, and there are restrictions on a measurer and a measurement place. EIA, FIA,
LIA, LPIA, etc. are advantageous in terms of handling safety, but use an optical system for final detection.
Components such as a lens, an aperture, and a light receiving element are required, and there are limitations on the weight and size of the measuring device. Also, EIA, FI
A, LIA, and the like often require a secondary reagent for coloring or emitting light, and the measuring device is complicated due to dispensing and washing of these reagents. In the case of the immunochromatography method, which is a simple measurement method, there is no special step in the reaction, the reaction proceeds only by adding a measurement sample or a diluent thereof, and the judgment is performed by a measurer visually.
It has the advantage of not requiring special equipment for measurement,
Since the determination is performed visually, accurate quantification of the measurement target is impossible, and there is a limitation that the obtained result is qualitative.

[0006]

Accordingly, as a result of intensive studies to solve this problem, particles containing a metal or a metal compound were used as a marker, and the amount of metal in the marker was measured for eddy current or capacitance. The inventors have found that the above problem can be solved by the measurement according to the present invention, and the present invention has been completed.

[0007] The present invention provides a method for immobilizing a specific binding substance that specifically binds biologically to an object to be measured, to a labeled particle containing a metal or a metal compound, through the specific binding substance, A method for measuring the amount of a measurement target in a sample, which comprises measuring the amount of metal of labeled particles bound to the measurement target by measuring eddy current or capacitance (hereinafter referred to as “the amount of metal bound to the measurement target”). , Also referred to as “the measurement method of the present invention”)
I will provide a.

[0008] In a preferred embodiment of the measurement method of the present invention, a stationary phase having at least one reaction site on which a specific binding substance that biologically specifically binds to an object to be measured is immobilized,
And, using a labeled particle containing a metal or a metal compound, in which a specific binding substance that biologically specifically binds to an object to be measured is immobilized, in the reaction site, The specific binding substance and the specific binding substance immobilized on the labeled particles are allowed to react with an object to be measured in a sample, and the amount of metal contained in the labeled particles bound to the reaction site by the reaction is reduced by eddy current or There is a method of measuring the amount of a measurement target in a sample, including measuring by measuring the capacitance.

In the measurement method of the present invention, the specific binding substance is preferably an antibody or an antigen.

[0010] The present invention also provides a membrane-like stationary phase having at least one reaction site on which a specific binding substance that specifically binds to an object to be measured is immobilized, Immobilized with a specific binding substance to be bound, a labeled particle-containing member containing a labeled particle containing a metal or a metal compound, wherein the labeled particle-containing member and the reaction site are on the test piece. A test piece (hereinafter referred to as a test piece) used for the measurement method of the present invention, the test piece being arranged such that the liquid containing the measurement object added to the sample flows in the stationary phase to move the labeled particles to the reaction site. , "The test piece of the present invention").

[0011] The present invention further provides a mounting portion for mounting a test piece including a stationary phase having at least one reaction site on which a specific binding substance that biologically specifically binds to an object to be measured is immobilized. A measuring device for measuring the amount of the object to be measured (hereinafter referred to as “the device of the present invention”) including a measuring portion for measuring the amount of metal contained in the reaction site of the test piece mounted on the mounting portion by measuring eddy current or capacitance. ").

In the apparatus of the present invention, the test piece is preferably the test piece of the present invention.

[0013]

BEST MODE FOR CARRYING OUT THE INVENTION The measuring method of the present invention is characterized in that a specific binding substance which specifically binds biologically to a measurement object is immobilized on a labeled particle containing a metal or a metal compound. The measurement target in the sample is bound via the substance, and the amount of metal of the labeled particles bound to the measurement target is measured by measuring eddy current or capacitance.

In the measurement method of the present invention, a labeled substance containing a metal or a metal compound is used as a labeled substance, and the detection of the labeled substance is performed by measuring the amount of metal in the labeled particle by measuring eddy current or capacitance. May be the same as a conventional measurement method based on the binding between a measurement object and a specific binding substance.

In the present invention, the object to be measured is not particularly limited as long as there is a specific binding substance that specifically binds biologically thereto. For example, examples of the combination of the measurement target and the specific binding substance include an antigen and an antibody, an antibody and an antigen, a sugar chain and a lectin, and a receptor and a ligand. The measurement object is preferably an antigen or an antibody that can be measured by an immunoassay using an antigen-antibody reaction. Examples of the antigen include various antigens such as proteins, polypeptides, steroids, polysaccharides, lipids, and viruses. That is, the antigen referred to in the present invention is, among all substances capable of inducing antibody production in humans or animals,
Refers to a single substance or a plurality of substances selected for a specific purpose such as diagnosis, or a mixture containing them.
Examples of the antibody include a protein that is an antibody against the above-mentioned antigen.

The labeled particles containing a metal or a metal compound (hereinafter, also referred to as “metal or metal compound-containing insoluble particles”) used in the present invention include, for example, metals such as iron, manganese, nickel, cobalt and chromium, cobalt, Alloys containing nickel, manganese, etc., triiron tetroxide (Fe
Metal or metal compound particles made of ₃ O ₄ ), iron sesquioxide (γ-Fe ₂ O ₃ ), various ferrites, or polystyrene, polymethyl methacrylate, polyacrylonitrile, polymethacryloyl containing these particles inside Nitriles, polycapramides, hydrophobic polymers such as polyethylene terephthalate, polyacrylamide, polymethacrylamide, polyvinylpyrrolidone, polyvinyl alcohol,
Poly (2-oxyethylene acrylate), Poly (2-
(Oxyethyl methacrylate), poly (2,3-dioxypropyl acrylate), poly (2,3-dioxypropyl methacrylate), crosslinked hydrophilic polymers such as polyethylene glycol methacrylate, or 2 to 4 types of each monomer Latexes such as copolymers, gelatin, liposomes, or particles in which the above-mentioned metal or metal compound particles are immobilized on the surface of latex, gelatin, liposomes, and the like are used.

The particle size of the metal or metal compound-containing insoluble particles is 0.05 μm to 5 μm, and 0.1 μm to 5 μm.
Those having a particle size in the range of m to 2 μm are preferred.

The content of the metal or metal compound of the metal or metal compound-containing insoluble particles is sufficient if the amount of metal of the particles bound to the object to be measured is sufficient to measure the eddy current or capacitance. It is often set as appropriate depending on the type of the object to be measured and the specific binding substance, the method of bonding the object to be measured and the particles, the means for measuring eddy current or capacitance, and the like. Usually, a metal or metal compound is used in an amount of 20 to 100% by mass, preferably 40% by mass, based on the whole particles.
It is preferred that the content be contained in an amount of from 80 to 80% by mass.

Insoluble particles containing a metal or a metal compound
Is used by immobilizing a specific binding substance. Special to immobilize
If the foreign binding substance is an antibody, the antigen to be measured
A human or animal-derived antibody capable of reacting with
can give. Antibodies include monoclonal antibodies and
Any of polyclonal antibodies may be used. Antibody to be immobilized
Is usually used, but pepsin, papaya
Digestive enzymes such as dithiothreitol, mercap
Using a reducing agent such as triethanol, F (ab ') _Two, F
ab ', Fab or the like having a low molecular weight may be used.
No. In addition, not only IgG but also IgM or IgG
A low molecular weight product obtained by the same treatment as described above may be used. one
On the other hand, in the case of antigen, for example, proteins, polypeptides,
Genetically engineered teroids, polysaccharides, lipids, viruses
Produced recombinant proteins, drugs, etc.
Reacts with antibodies from elephant, e.g., human or animal
Materials that can be used are used.

As a method for supporting a specific binding substance, for example, an antibody or an antigen, on a metal or a metal compound-containing insoluble particle, a specific binding substance is bound to a labeled particle in a conventional assay using a biologically specific reaction. And the methods used to do so. For example, it is carried out by physically adsorbing them or chemically bonding them.

Examples of the physical adsorption method include a method of directly immobilizing a specific binding substance on particles, and a method of chemically adsorbing and immobilizing the specific binding substance to another protein such as albumin. As a method of chemically supporting, the amino group, carboxyl group, mercapto group, hydroxyl group, aldehyde group,
A method of immobilizing directly on particles using a functional group that can bind to a specific binding substance molecule by chemically modifying an epoxy group, etc., a spacer molecule between the particle and the specific binding substance molecule And a method of chemically bonding a specific binding substance to another protein such as albumin and then chemically bonding the protein to particles.

The particles to which the specific binding substance has been immobilized can be used to prevent non-specific binding of the substance to be measured to the particles in the same manner as the labeled particles in the conventional measurement method based on the binding between the substance to be measured and the specific binding substance. To prevent this, treatment with an aqueous bovine serum albumin solution or the like may be used.

The measurement of the metal content of the labeled particles by measuring the eddy current or the capacitance can be performed by a method known per se. For example, there is a method using an eddy current type or capacitance type displacement sensor described later.

The assay method of the present invention preferably comprises a stationary phase having at least one reaction site on which a specific binding substance that biologically specifically binds to an object to be measured is immobilized;
Using a labeled particle containing a metal or a metal compound, in which a specific binding substance that biologically specifically binds to an object to be measured is immobilized, the specific site immobilized in the reaction site in the reaction site. The binding substance and the specific binding substance immobilized on the labeled particles are allowed to react with the measurement target in the sample, and the amount of metal contained in the labeled particles bound to the reaction site by the reaction is reduced by eddy current or electrostatic force. This is a method for measuring the amount of a measurement target in a sample, including measuring by measuring the volume.

The carrier used as the stationary phase in the reaction may be a nitrocellulose membrane, a glass filter membrane, a nylon membrane, a modified nylon membrane, a polyvinylidene difluorolide (PVDF) membrane or other membrane-like material, a filter paper,
Non-woven fabrics (for example, cellulosic fibers such as rayon and cotton yarn,
Sheet materials such as glass fibers or those prepared from chemical fibers such as nylon, polyester, polypropylene, and polyurethane).

A specific binding substance such as an antibody or an antigen is immobilized on the reaction site of the stationary phase. The immobilization on the stationary phase can be performed in the same manner as the immobilization on the metal or metal compound-containing insoluble particles described above. The specific binding substance immobilized on the reaction site may be the same as or different from the specific binding substance immobilized on the particles.

As a method of reacting a specific binding substance immobilized on particles with an object to be measured, a sample (a diluent thereof if necessary) and labeled particles are mixed in advance in a test tube, a plastic tube, a microplate, or the like. The reaction may be carried out, or may be carried out simultaneously with moving the surface or inside of the stationary phase.

When the specific binding substance immobilized on the particles has been reacted with the analyte in advance, a part of the stationary phase is immersed in a mixture of the sample (if necessary, a diluent thereof) and the labeled particles. By the flow of solution penetrating by capillary action,
The labeled particles to which the analyte is bound can be moved to the reaction site where the specific binding substance is immobilized through the surface or inside the stationary phase. In the case of simultaneous movement when moving on the surface or inside of the stationary phase, a member containing labeled particles in which a specific binding substance is immobilized is arranged at one end of the stationary phase, bonded in advance, and measured. At this time, the sample (the diluent, if necessary) is allowed to penetrate into the labeled particle-containing member, thereby causing the labeled particles to flow and reacting the specific binding substance bound on the particles with the analyte in the sample. ,
The particles can be moved to a reaction site on the stationary phase. At this time, the member containing the labeled particles includes glass fiber, woven fabric, non-woven fabric and the like.

The mixture of the labeled particles and the sample (a diluent, if necessary) moves on the surface or inside of the stationary phase, and moves to the reaction site, whereupon the specific binding substance immobilized on the stationary phase is measured. The analyte or the analyte bound to the labeled particles, or the analyte bound to the stationary phase reacts with the specific binding substance bound on the labeled particles (ie, immobilized at the reaction site). The specific binding substance and the specific binding substance immobilized on the label particles react with the measurement target in the sample), and a part of the migrated label particles remains (binds) at the reaction site, The remaining labeled particles move further past the reaction site.

The labeled particles remaining at the reaction site of the stationary phase are
It is measured as the amount of metal contained therein by measuring eddy current or capacitance. As a method of measuring the metal amount by measuring the eddy current or the capacitance, for example, a method using an eddy current type or a capacitance type displacement sensor (also referred to as a proximity sensor, a linear sensor, or the like) is exemplified. The principle of the eddy current displacement sensor is that a high-frequency current is applied to the coil inside the tip of the detector to generate a high-frequency magnetic field. In this case, the eddy current in the vertical direction flows through the magnetic flux and the impedance of the internal coil of the detector changes, and the distance from the tip of the detector to the object to be measured is measured by the change in the transmission state due to this phenomenon (for example, EX-V series manufactured by Keyence). Capacitive displacement sensors are
This is based on the principle that the capacitance of a flat plate capacitor is inversely proportional to the distance between the electrodes.Changes in the capacitance that occur on the surface of the detector tip and the surface of the object to be measured are converted to electrical signals by a bridge circuit and measured. By doing so, the distance from the tip of the detector to the object to be measured is measured (for example, Z4KC made by OMRON). These sensors are usually used for measuring a metal lump as a measurement target and detecting a minute change in the position thereof in a non-contact manner. However, the same principle can be used to quantify the amount of metal present in a certain range. If the distance from the tip of the detector to the object to be measured is kept constant and the metal particles are sufficiently small with respect to the area of the measurable region, a signal proportional to the amount of the metal (the number of particles) can be obtained. In actual measurement, the tip of the detector is brought close to or in contact with the surface of the reaction site of the stationary phase where the reaction has been completed, and the signal (voltage output, current output, etc.) from the sensor is detected, and the residual The measured amount of metal is measured, from which the amount of the object to be measured is obtained.

In the quantification of the object to be measured, a calibration curve relating to the concentration of the object to be measured and the output signal of the sensor is stored in the measuring device in advance, or a calibration curve is prepared by actually measuring a standard product having a known concentration. The concentration of the measurement object can be calculated from the measured value of the measurement sample and the calibration curve. A standard site for measuring the total amount of the labeled particles flowing on the stationary phase may be provided in addition to the reaction site of the stationary phase, and the calibration curve may be corrected by measuring the amount of metal at the standard site.

Further, the correction by the lot difference in the production of the stationary phase and the labeled particles is performed by storing the shape of the calibration curve in the measuring device in advance, performing the target measurement at the time of measurement, and using the result of the original calibration curve (master). Curve), or write the calibration curve information on a test piece having a stationary phase and labeled particles using a barcode or the like, read it out to a measuring device, and correct the original calibration curve. It can be carried out. The latter method is preferable when applied to a simple inspection because measurement for correction is unnecessary when performing the inspection.

The test piece of the present invention is a test piece to be used in the measurement method of the present invention, and is a film-like immobilization having at least one reaction site on which a specific binding substance that specifically binds to an object to be measured is immobilized. Phase, and a labeled particle-containing member containing labeled particles containing a metal or a metal compound, in which a specific binding substance that biologically specifically binds to the measurement object is immobilized, and the labeled particle-containing member is included. And the reaction site is
The liquid containing the object to be measured applied to the test piece is arranged so as to flow the labeled particles in the stationary phase and move to the reaction site.

The carrier used as the membrane stationary phase is the same as the stationary phase described above. Preferably it is porous. Reaction sites can be created as described above.

The member containing the labeled particles to which the specific binding substance is immobilized is usually placed at one end of the membrane stationary phase and bonded (FIG. 1). In this case, examples of the member containing the labeled particles include glass fiber, woven fabric, and nonwoven fabric.

At the time of measurement, a liquid (analyte or a diluent thereof) containing an object to be measured is caused to penetrate the labeled particle-containing member (3), thereby causing the labeled particles to flow and the specific binding substance bound on the particles. The particles can be moved to the reaction site (2) on the film-like stationary phase (1) while reacting the sample and the measurement object in the sample. The mixture of the labeled particles and the sample or the diluent thereof moves on the surface or inside of the membrane-like stationary phase (1), and moves to the reaction site (2), whereupon the specific immobilized on the reaction site (2) is obtained. The binding substance and the specific binding substance immobilized on the label particles react with the measurement target in the sample, and a part of the migrated label particles remains (binds) at the reaction site (2) and remains. Are adapted to move further through the reaction site.

The apparatus of the present invention comprises: a mounting portion for mounting a test piece including a stationary phase having at least one reaction site on which a specific binding substance that biologically specifically binds to an object to be measured is immobilized; A measurement unit is provided for measuring the amount of metal contained in the reaction site of the test piece mounted on the mounting unit by measuring eddy current or capacitance. The measurement method of the present invention can be performed using the device of the present invention.

The test piece mounted on the mounting portion is a test piece containing a stationary phase as described in the measurement method of the present invention. The mounting part is adapted to mount such a test piece. The test piece is preferably a test piece of the present invention.

The measuring section includes, for example, an eddy current or capacitance sensor such as the above-mentioned eddy current type or capacitance type displacement sensor. The device of the present invention preferably further includes a storage unit for storing information on the calibration curve, and a calculation unit for calculating the amount of the measurement target from the information stored in the storage unit and a measurement value by the measurement unit. Further, the apparatus may further include an input device for inputting information of the calibration curve, for example, a keyboard, a barcode reader, and the like. Preferably, an input device for reading information written on the test piece by a barcode or the like, and a calculation unit for correcting a calibration curve based on the information read by the input device are preferable.

[0040]

EXAMPLES Hereinafter, the present invention will be described in detail with reference to examples, but the present invention is not limited to the following examples unless it exceeds the gist. In addition,% is mass% unless otherwise specified.

[0041]

Example 1 Human IgM Measurement (1) Iron oxide-containing polystyrene latex having an average particle diameter of 1.387 μm (Ceradyne, iron oxide content: 60%)
Was added to 2 ml of a 1% suspension of a 1% suspension of a monoclonal antibody against human IgM (Medix Biochemical, Clone No. 7410) and 20 mg of carbodiimide to thereby immobilize the monoclonal antibody on the latex by a chemical bonding method. Stabilize the particles by treatment with albumin and add 0.1%
To prepare a metal compound particle-labeled antibody reagent.

(2) A nitrocellulose membrane (Whatman) having a pore size of 8 μm was cut into strips of about 6 mm × 40 mm, and a monoclonal antibody against human IgM (Medix Biochemical, Clone No. 7408) was placed almost at the center. Was added dropwise at a concentration of 0.5 mg / ml, air-dried, immersed in bovine serum albumin solution, and air-dried to prepare a stationary phase having a reaction site at the center.

(3) Three nitrocellulose membranes as the stationary phase are prepared, and one end of the strip is provided with 0 m of human IgM (Jackson ImmunoResearch) as a measurement sample.
g / ml (buffer only), 0.05 mg / ml, 0.5
50 μl of each solution having a concentration of mg / ml was dropped.

(4) On the same site where the measurement sample was dropped, 0.1% of the above-mentioned polystyrene latex containing iron oxide, on which a monoclonal antibody against human IgM was immobilized.
% Suspension was added dropwise, and latex particles were moved on a nitrocellulose membrane while reacting. Several minutes later, it was visually observed that the iron oxide-containing polystyrene latex remained at the central reaction site of the two films on which the measurement sample containing human IgM was dropped.

(5) The tip of the detector of the eddy current displacement sensor (Keyence Corporation) was brought into contact with the reaction site at the center of the nitrocellulose membrane where the reaction was completed, and the output voltage was measured. The results are shown in FIG. Human IgM in the measurement sample
As the concentration increased, a decrease in the voltage value was observed.

[0046]

Example 2 Measurement of Human IgM (1) Iron oxide-containing polystyrene latex having an average particle diameter of 1.387 μm (Ceradyne, iron oxide content: 60%)
Was added to 2 ml of a 1% suspension of a 1% suspension of a monoclonal antibody against human IgM (Medix Biochemical, Clone No. 7410) and 20 mg of carbodiimide to thereby immobilize the monoclonal antibody on the latex by a chemical bonding method. The particles were stabilized by treating with albumin, and suspended in a buffer at a latex concentration of 1% to prepare a metal compound particle-labeled antibody reagent.

(2) A nitrocellulose membrane having a pore diameter of 8 μm (Whatman) was cut into a strip of about 4 mm × 35 mm, and a monoclonal antibody against human IgM (Medix Biochemical, Clone No. 7410) was placed at the approximate center. After dropping 1 μl at a concentration of 1 mg / ml and air-drying, it was immersed in bovine serum albumin solution and air-dried to prepare a stationary phase having a reaction site at the center.

(3) A glass fiber membrane (conjugate release pad,
(Whatman) was cut into a size of 5 mm × 5 mm, and 4 μl of a 1% suspension of the anti-human IgM monoclonal antibody-sensitized latex prepared in (1) was dropped thereon to prepare a labeled particle-containing member. The labeled particle-containing member was overlaid on one end of the nitrocellulose membrane as the stationary phase prepared in (2) to obtain a test piece.

(4) Six test pieces were prepared, and 0 mg / ml (buffer only) of human IgM (Kappel) and 0.13 mg /
dl, 0.27 mg / dl, 0.54 mg / dl, 1.
50 μl of solutions having a concentration of 07 mg / dl and 2.15 mg / dl were respectively added dropwise.

(4) By dropping 50 μl of the buffer solution continuously onto the same site where the measurement sample was dropped, the latex immobilized with the anti-human IgM monoclonal antibody held by the labeled particle-containing member was placed underneath. Transferred onto a nitrocellulose membrane, and
It was moved horizontally on a nitrocellulose membrane while reacting with gM. After a few minutes, it is visually observed that the iron oxide-containing polystyrene latex remains at the central reaction site of the film on which the measurement sample containing human IgM was dropped, and the measurement sample containing no human IgM (buffer solution) ) Was not observed.

(5) The tip of the detector of the capacitance type displacement sensor (OMRON) was brought into contact with the reaction site at the center of the nitrocellulose membrane after the reaction was completed from the back side, and the output voltage was measured. Show. As the concentration of human IgM in the measurement sample increased, a decrease in the voltage value was observed.

[0052]

According to the method of the present invention for measuring the metal content of the labeled particles by measuring the eddy current or the capacitance, the detection can be performed only by bringing the sensor close to a container or a carrier where a reaction occurs. A sensor for measuring the amount of metal of this type, for example, an eddy current detection sensor, can be reduced in weight and size, and the measurement device can be simplified because no secondary reagent is required for detection. In addition, if the present method is applied to an immunochromatography method, a simple measuring method can be provided that does not require a special device for the reaction and can further quantify an object to be measured.

[Brief description of the drawings]

FIG. 1 shows a schematic diagram of an example of a test piece.

FIG. 2 shows the relationship between the concentration of human IgM in a sample and the output voltage of an eddy current displacement sensor obtained in Example 1.

FIG. 3 shows the relationship between the concentration of human IgM in a sample and the output voltage of a capacitance displacement sensor obtained in Example 2.

[Explanation of symbols]

 DESCRIPTION OF SYMBOLS 1 Membranous stationary phase 2 Reaction site 3 Labeled particle containing member

Claims (6)

[Claims] 1. A method for measuring in a sample via a specific binding substance, a labeled particle containing a metal or a metal compound, on which a specific binding substance that biologically specifically binds to an object to be measured is immobilized. A method for measuring the amount of an object to be measured in a specimen, comprising binding an object and measuring the amount of metal of the labeled particles bound to the object by measuring eddy current or capacitance. 2. The method according to claim 1, wherein a reaction site on which a specific binding substance that biologically specifically binds to the measurement target is immobilized is at least one.
Using a labeled particle containing a metal or a metal compound, in which a specific binding substance that biologically specifically binds to a measurement object is immobilized, and the reaction site, Reacting the immobilized specific binding substance and the specific binding substance immobilized on the labeled particles with a measurement object in a sample, and reacting the metal contained in the labeled particles bound to the reaction site by the reaction; Measuring the amount by measuring eddy current or capacitance,
A method for measuring the amount of a measurement target in a sample. 3. The method according to claim 1, wherein the specific binding substance is an antibody or an antigen. 4. A membrane-like stationary phase having at least one reaction site on which a specific binding substance that specifically binds to a measurement target is immobilized, and a specific binding phase that specifically binds biologically to the measurement target. A test piece including a labeling particle-containing member containing labeling particles containing a metal or a metal compound in which a binding substance is immobilized, wherein the labeling particle-containing member and the reaction site are added on the test piece. A liquid containing an object to be measured is a test piece arranged to flow the labeled particles in a stationary phase and move to a reaction site, and is used in the method according to any one of claims 1 to 3. Specimens for 5. A reaction site on which a specific binding substance that biologically specifically binds to an object to be measured is immobilized.
A mounting part for mounting a test piece including a stationary phase having a plurality of stationary phases, and a measuring part for measuring the amount of metal contained in the reaction site of the test piece mounted on the mounting part by measuring eddy current or capacitance. A device for measuring the quantity of an object to be measured. 6. The apparatus according to claim 5, wherein the test piece is the test piece according to claim 4.