JP2001147226A - Dispensing instrument having reagent component arranged thereto - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide an dispensing instrument capable of omitting the number of processes than before and minimizing the necessary use amounts of a liquid and a reagent when a substance present in a liquid and a reagent are reacted and the reagent is dissolved or suspended in the liquid and a reaction method of the substance in the liquid and the reagent using this instrument. SOLUTION: In the dispensing instrument sucking a liquid to discharge a definite amount of the liquid, a reagent is arranged in this instrument. In a reaction method of a substance in a liquid and the reagent, the liquid is sucked to be housed in the instrument and the reagent arranged in the instrument is dissolved or suspended in the liquid or mixed with the liquid to be reacted with the substance in the liquid.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a dispensing device in which reagent components are arranged, and a method for reacting a substance in a liquid with a reagent component using the dispensing device.

[0002]

2. Description of the Related Art Conventionally, when reacting a substance present in a liquid with a component of a reagent, the following procedure is followed. 1. Aspirate or weigh reagent components into dispensing device. 2. The components of the one reagent are stored in a container. 3. Aspirate water, buffer or organic solvent into dispensing device. 4. A certain amount of the sucked water or the like is discharged to a container in which the components of the above two reagents are stored. Thereby, the components of the reagent are dissolved or suspended. 5. The liquid is drawn up by the dispensing device. 6. A certain amount of the sucked liquid is discharged into the container (4). At this time, the reaction between the substance present in the liquid and the components of the reagent is performed in this container. 7. Next, the solution or suspension in which the reaction has been performed is sucked up by a dispensing device. 8. Dispense a certain amount of this solution or suspension. 9. For this discharged fixed amount of solution or suspension,
Perform the next operation.

[0003] When a reagent component is dissolved or suspended in a liquid, the following procedure has conventionally been used. 1. Aspirate or weigh reagent components into dispensing device. 2. The components of the one reagent are stored in a container. 3. Aspirate liquid into dispensing device. 4. A certain amount of the sucked liquid is discharged to a container containing the components of the reagent. Thereby, the components of the reagent are dissolved or suspended in the liquid. 5. Next, the solution or suspension is sucked up by the dispensing device. 6. Dispense a certain amount of this solution or suspension. 7. For this discharged fixed amount of solution or suspension,
Perform the next operation.

As described above, conventionally, both in the case of reacting a substance present in a liquid with a reagent component and in the case of dissolving or suspending a reagent component in a liquid, a large number of steps are required. There is a problem that the components of the reagent are left unnecessarily.

[0005]

DISCLOSURE OF THE INVENTION The present invention requires a smaller number of steps than in the prior art when reacting a substance present in a liquid with a reagent component and when dissolving or suspending the reagent component in a liquid. It is an object of the present invention to provide a dispensing device which can be omitted and requires a minimum amount of liquid and reagent to be used, and a method of reacting a substance in a liquid with a component of the reagent using the dispensing device.

[0006]

(1) A liquid is sucked up,
A dispensing device for discharging a certain amount of the liquid, wherein a component of a reagent is disposed therein. (2) The dispensing device according to (1), wherein the reagent is a measurement reagent. (3) The dispensing device according to the above (1) or (2), wherein a reaction between a substance in the sucked-up liquid and a reagent component is performed in the dispensing device. (4) After the components of the reagent arranged in the dispensing device are dissolved, suspended or mixed by the sucked liquid, or after reacting with the substance in the sucked liquid, a certain amount of the reagent is dispensed from the dispensing device. The dispensing device according to any one of (1) to (3), which is discharged.

(5) The dispensing device described in (1) above sucks up the liquid and dissolves, suspends, or mixes the components of the reagent arranged in the dispensing device. A method of reacting a substance in a liquid with a reagent component, wherein the reagent component is reacted with the reagent in the dispensing device. (6) The method according to (5), wherein the reagent is a measurement reagent. (7) The method according to the above (5) or (6), wherein a certain amount of a reaction between the substance in the liquid and the reagent component in the dispensing device is discharged from the dispensing device.

[0008]

BEST MODE FOR CARRYING OUT THE INVENTION A dispensing device according to the present invention is a dispensing device for sucking up a liquid and discharging a fixed amount of the liquid.
It is characterized in that components of the reagent are arranged therein. In the dispensing device of the present invention, “reagent components” to be disposed on the dispensing device are chemical components used for a specific purpose of use. The “components of the reagent” may be one kind or plural kinds, and may be arranged at one place in the dispensing device or arranged at plural places. The components of the reagent may be arranged in the dispensing device, or may be impregnated in a carrier or the like.

The "components of the reagent" include, for example, the components of the measurement reagent, but are not limited thereto, and may be components of the reagent other than the components of the measurement reagent. The `` component of the measurement reagent '' is a component of a reagent used for measurement of an object to be measured, for example, a component of a biochemical reaction measurement,
Examples include components for measuring an immunological reaction, components for measuring a blood coagulation time, and components for a nucleic acid probe method. Examples of the "immunological measurement reaction" include an immunotrap method (Japanese Patent Application Laid-Open No. 8-63733), an immunoturbidimetric method (TIA), a latex turbidimetric method, an indirect agglutination reaction, an enzyme immunoassay (EIA) ), Fluorescence immunoassay (FIA), radioimmunoassay (RIA), luminescence immunoassay (LIA),
Examples include an immunochromatography method. Examples of the “components of the reagent other than the measurement reagent component” include a protein removing agent (for example, used as a pretreatment agent for measurement), a detergent (for example,
(Used for washing the measurement container).

[0010] The "components of the reagent" may be solid or liquid. When the “reagent component” is solid, the “reagent component” may be dissolved or suspended in a liquid. If the "reagent component" is liquid, the "reagent component"
May be concentrated. When the “reagent component” is in a liquid state, the “reagent component” may be discharged into a container, and the “reagent component” and the liquid may be mixed in the container. The amount of the “reagent component” disposed in the dispensing device of the present invention depends on the purpose of the reagent.
It may be a batch or a plurality of batches.

[0011] The dispensing device of the present invention includes, for example, a pipette,
A pipette of an analyzer or a liquid collection / dispensing device such as a tip of these pipettes, or another dispensing device such as a dropper bottle. The dispensing device of the present invention can be used in any operating method, such as a manual method or an automated method. The dispensing device of the present invention is provided with a wall on the side of the outlet of the reagent component disposed in the dispensing device (or further on the opposite side) to prevent leakage of the reagent component. It may be. In the dispensing device of the present invention, the outlet may be narrowed and narrowed, or the outlet may be sealed. The dispensing device of the present invention can dissolve, suspend or mix components of a reagent in a liquid in the dispensing device. The dispensing device of the present invention can cause a reaction between a substance in a liquid and a reagent component in the dispensing device.

According to the method of the present invention for reacting a substance in a liquid with a reagent component, the above-described dispensing device sucks up the liquid and dissolves, suspends, or mixes the reagent component disposed in the dispensing device. Thereby, the substance in the liquid and the component of the reagent are reacted in the dispensing instrument. When the components of the reagent are in a solid form, they are dissolved or suspended in a liquid, and when in a liquid form,
This is mixed with the liquid.

The temperature of the reaction may be selected to be suitable for the reaction. In general, the higher the temperature, the higher the reaction rate. However, the higher the temperature, the higher the risk of deactivation or decomposition of the substance or reagent component in the liquid. Therefore, it is necessary to carry out the reaction in a temperature range in which these are not deactivated or decomposed. The reaction time may be selected according to the purpose of the reaction. After the reaction is performed, a certain amount of the solution or suspension in which the reaction has been performed may be discharged and used for the next operation.

The components of the present invention will be described below. 1. Reagent Components Reagent Components In the present invention, “reagent components” are chemical components used for a specific purpose of use. Examples of the components involved in the target chemical reaction include components of the measurement reagent, components of the reagent other than the components of the measurement reagent, and the like. The component of the measurement reagent is a component used for measuring a measurement target.

[0015] What is used for measuring a biochemical reaction is as follows:
For example, first reagent, second reagent, third reagent, fourth reagent, substrate, enzyme, coenzyme, buffer, chromogen, stabilizer (eg, HSA, BSA, casein, PVP, SH compound), catalyst , A reactant, an activator, an activator, an inhibitor, an inhibitor, a metal, a chelating agent, an SH group-containing material, an oxidant, and a reducing agent.

Among the immunological reaction measurements, those used in the immunotrap method (the method described in JP-A-8-63733) include, for example, antibody-bound particles, antigen-bound particles, antibody-bound magnetic particles, and antigen-bound particles. Magnetic particles. Antibodies used for immunoturbidimetry include, for example, antibodies. Examples of the latex turbidimetric method include antibody-bound latex particles and antigen-bound latex particles. Indirect agglutination reaction (for example, RPHA method (reversed
passive agglutination), PHA method (passive agglutina
tion), PA method (particle agglutination), MAT (mag
netic agglutination test) (JP-A-4-216466,
JP-A-4-242167), SPAT method (solid
phase agglutination test) (the method described in JP-A-64-69954), for example, antibody-bound red blood cells, antigen-bound red blood cells, antibody-bound particles, antigen-bound particles, antibody-bound magnetic particles, antigen-bound magnetic particles Is mentioned. Examples of those used for EIA, FIA, RIA, or LIA include enzyme-labeled antibodies, enzyme-labeled antigens, fluorescent-labeled antibodies, fluorescent-labeled antigens, radioactive-labeled antibodies, radioactive-labeled antigens, luminescent-labeled antibodies, and luminescent-labeled antibodies Examples include antigens, enzyme-labeled antibody-bound particles, enzyme-labeled antigen-bound particles, substrates, and chromogens. Examples of those used in the immunochromatography include colloidal gold-labeled antibodies, colloidal gold-labeled antigens, colored latex particle-labeled antibodies, and colored latex particle-labeled antigens.

As the one used for measuring the blood coagulation time, for example, activated partial thromboplastin time (APT)
T) Measurement reagent, partial thromboplastin time (PTT)
Measuring reagent, tissue thromboplastin / calcium mixture,
Thrombotest reagent, platelet suspension, sodium citrate, calcium chloride; for use in nucleic acid probe methods, for example, nucleotides, labeled nucleotides;
Examples of those used for lectin reaction measurement include sugar,
Lectin, labeled sugar, labeled lectin;
Receptors, labeled ligands, and labeled receptors.

[0018] Other substances used for measurement utilizing reactions include, for example, covalent bond reactants, hydrogen bond reactants, positively or negatively charged substances in ionic bond reactions, hydrophobic substances in hydrophobic bond reactions, and adsorbent bonds. Reactants. Examples of the reagent components other than the measurement reagent component include a pretreatment agent (for example, a protein-removing agent), a specific binding component (for example, a component used for PCR), a detergent (for washing a measurement container), and the like.

Reagent Component States The reagent components may be in a solid state (for example, one in which the reagent component is dissolved in a liquid, one in which the reagent component is suspended in a liquid), or a liquid state (for example, a reagent component). One in which the components are not concentrated or one in which the components of the reagent are concentrated).

2. Dispensing device The dispensing device of the present invention is a dispensing device for sucking up a liquid and discharging a certain amount of the liquid, wherein components of the reagent are arranged therein. Dispensing device The dispensing device includes a liquid sampling and dispensing device and other dispensing devices.

Examples of the liquid collecting / dispensing device include a pipette, a micropipette, a pipette of an analyzer, a pipette of a diluting device, a pipette of a dispensing device, a pipette of a diluting / dispensing device, a pipette of a washing device, and a tip of a micropipette. Examples include a pipette tip of an analyzer, a pipette tip of a diluting device, a pipette tip of a dispensing device, a pipette tip of a dilution dispensing device, a pipette tip of a washing device, a dropper, a dropper, and a capillary (capillary).

Examples of the pipette include a whole pipette, an Eppendorf type pipette, a Komagome pipette, an Ostwald pipette, and a Pasteur pipette. Examples of these materials include glass, synthetic resin, ceramics, and Teflon (registered trademark). The volume is not limited, but is usually 0.01 μL to 100 mL. These pipettes are filtered pipettes,
That is, the pipette may be a pipette in which a filter made of a material such as hydrophobic polyethylene is installed,
A tapered pipette, that is, a pipette whose outlet is narrowed and narrowed may be used. The material, volume and the like of the filter-containing pipette and the tapered pipette may be the same as described above. Examples of tapered pipette products include Funakoshi's ARTGEL, TOHO's gel saver, and Assist's transfer pipette.

A micropipette is a pipette capable of sucking up or discharging a liquid by operating a push button or operating (moving) a knob. Although the volume is not limited, it is usually 0.1 μL to 20 mL, and any of a fixed volume type, a multiple volume selection type, and a continuous volume variable type may be used.
The channel may be single-channel type or multi-channel type, the dispensing method may be single-dispensing method or continuous dispensing method, and the power is manual, automatic (battery,
Electricity). Examples of micropipette products include Eppendorf Reference 4910, Reference 4900, Research 3110, 3190, Titerman 4908,
Biomaster 4830, 4720, Multipet Plus 498
0, 4780, Plus / 8, 5221; Gilson Pipetman; Nichiryo Autoclavable Bench Mate,
Benchmate F & V, Juster 1100DG, Juster F & V
, MODEL 800, MODEL 7000, MODEL 8100, MODEL 880
0, chip master.

Examples of the material of the tip of the micropipette include synthetic resin (eg, polypropylene), glass, ceramics, and Teflon. Although the volume is not limited, it is usually 0.01 μL to 100 mL, for example, 2 μL.
L, 2.5 μL, 10 μL, 20 μL, 100 μL, 2
00 μL, 1000 μL, 1.25 mL, 2.5 mL,
5.0 mL. Examples of micropipette tip products include Eppendorf Stander Tip, Euro Tip, BioPure Tip, Master Tip, Bali Tip Combi Tip Plus, Combi Tip Plus Biopure; Nichiryo Bench Mate Tip; Sorenson Multipipette Tip, Scale Multi tip, multi micro tip; micro pipette tip by Assist Co .; pipette tip for quick turbo by Sinotest Co., Ltd.

The tip of the micropipette may be a filter tip, that is, a filter tip in which a filter made of a material such as hydrophobic polyethylene is installed in the tip, or a tapered tip, that is, a discharge port is narrowed. Thinner chips may be used. Examples of filter chip products include Eppendorf's filter tip; Nichiryo's clean tip; Sorenson's multi-guard filter tip, multi-micro guard tip, multi-guard filter tip (E), ART pipette tip, AR
T Ultramicro pipette tips and ART activated carbon filter pipette tips. Examples of the product of the tapered tip include a Geloda chip manufactured by Eppendorf; a multiflex chip, a multiminiflex chip, an Ecoflex microcapillary chip manufactured by Sorenson; and a capillary chip for a quick turbo manufactured by Sinotest. Other dispensing devices include, for example, drop bottles.

Aspects of the dispensing device of the present invention The dispensing device of the present invention may be provided with a wall for preventing leakage of the reagent components on the side of the outlet of the reagent components disposed therein. . Further, a similar wall may be provided on the opposite side (on the opposite side of the disposed components of the reagent). This can prevent the arranged reagent components from leaking out of the dispensing instrument, and prevent the reagent components from entering the liquid and contaminating the liquid when sucking up the liquid. The material of this wall is a porous material, and the size of the pores is such that the components of the reagent before dissolution or suspension do not leak out and the components of the reagent after dissolution or suspension pass through. There is no particular limitation as long as it is possible, for example, cellulose acetate, modified polyvinylidene fluoride, nylon,
Examples include polytetrafluoroethylene (Teflon) and polyvinylidene fluoride. As a method of providing a wall in the dispensing device, for example, from the opposite side of the discharge port of the dispensing device,
A method of packing the material formed into an appropriate size is exemplified. In some cases, the wall may be adhered to the dispensing device with an adhesive.

The size of the pores in the wall must be such that the components of the reagent before dissolution or suspension do not leak out and that the components of the reagent after dissolution or suspension can pass through. The size of the components of the reagent in the state of being dissolved or suspended in a liquid (usually the size of the molecules of the components of the reagent) must be smaller than the size of the pores in the wall.

In this case, in the state of being dissolved or suspended in the liquid, the components of the reagent are dissolved or suspended (dispersed) in the liquid in a state of one molecule or close to the molecule, so that the reagent can pass through the hole in the wall. . On the other hand, when the reagent is placed in the dispensing device in a dry state, the individual components of the reagent are collected to form a solid and cannot pass through the hole in the wall.

In the dispensing device of the present invention, the discharge port may be narrowed and narrowed. This also prevents the arranged components of the reagent from leaking out of the dispensing instrument, and prevents the components of the reagent from entering the liquid and contaminating the liquid when sucking up the liquid. As a method of narrowing the discharge port of the dispensing instrument, for example, heat molding can be mentioned.

In order to prevent the arranged reagent components from leaking out of the dispensing instrument by narrowing the discharge port, it is necessary to dissolve or suspend the reagent in a liquid. The size of the reagent component (usually the size of the molecule of the reagent component) must be smaller than the size of the outlet of the dispensing instrument. In this case, in a state of being dissolved or suspended in the liquid, the components of the reagent are dissolved or suspended (dispersed) in the liquid in a state of one molecule or close to one molecule, and thus can pass through the discharge port.

On the other hand, when the reagent is placed in the dispensing apparatus in a dry state, the individual components of the reagent are gathered to form a solid and cannot pass through the discharge port. The components of the reagent to be arranged in the dispensing device may be arranged at one place in the dispensing device, or may be arranged at a plurality of places.

The components of the reagent to be arranged in the dispensing device may be one kind or a plurality of kinds. For example, a carrier containing a reagent component different from the disposed reagent component may be provided on the side of the outlet of the reagent component disposed in the dispensing instrument. The carrier must be able to pass through the components of the reagent after dissolution or suspension (for this, see the description of the wall provided in the dispensing device described above). For example, a substance capable of absorbing a substance causing a non-specific reaction in the carrier (for example, an antibody against a substance causing a non-specific reaction) is contained in the carrier, so that the substance contained in the sucked liquid is contained. A substance that causes a specific reaction can be absorbed and removed, and thereafter, a component of the arranged reagent can react with a liquid that has absorbed and removed the substance that causes a non-specific reaction. Further, by storing a salt or a buffering agent having a buffering capacity in a high pH range or a low pH range on the carrier, the components of the arranged reagent can withstand a low salt concentration, or a low pH or a high pH. Even if it is not possible, the reaction between the sucked-up liquid and the component of the reagent placed can be performed under conditions of high salt concentration, high pH or low pH. Further, a carrier containing a substance (for example, an antibody or an antigen) capable of specifically binding to a measurement target (for example, an antigen or an antibody) in the liquid is provided, and the measurement is performed using the carrier. A confirmation test for confirming whether or not the object to be measured truly exists can be performed.

The amount of the reagent component to be placed in the dispensing device is
The reagent may be used once or multiple times. The outlet of the dispensing instrument may be sealed. To seal, the outlet is covered with a stopper or cap, filled with clay or putty, wrapped with tape,
Alternatively, it can be performed by attaching a seal or the like. Also,
The discharge port may be thinned, crushed, or bonded or fused. In this case, the discharge port is used by expanding or cutting the discharge port.

The components of the reagent to be arranged in the dispensing device may be arranged in the dispensing device with the reagent component itself, or may be impregnated in a carrier or the like. Examples of the method for disposing the components of the reagent in the dispensing device include freeze-drying, air-drying, impregnation, packing, and siphoning.

In freeze-drying, usually, a fixed amount of a liquid component of a reagent is sucked into a dispensing device, and a freeze-drying process is performed to dispose the reagent component on the dispensing device. Note that it is preferable that the freeze-drying process be performed using liquid nitrogen, because the solution can be instantly frozen and the components of the reagent can be uniformly arranged in the dispensing instrument.

In air-drying, usually, a certain amount of a liquid reagent component is sucked into a dispensing device, and dried at room temperature or before or after the same without blowing or blowing, whereby the reagent component is placed in the dispensing device. I do.

In the impregnation, usually, a fixed amount of a liquid reagent component is transferred to a water-absorbing carrier (cellulose, nonwoven fabric, sponge, etc.).
This is carried out by placing the sample in a dispensing device or impregnating the water-absorbing carrier placed in the dispensing device with a certain amount of a liquid reagent component.
In some cases, the components of the impregnated reagent are freeze-dried or air-dried.

In packing, usually, a fixed amount of a solid component of a reagent is packed in a dispensing device, so that the reagent component is arranged in the dispensing device. In the siphoning, usually, a fixed amount of a liquid component of the reagent is sucked into the dispensing device, so that the reagent component is arranged in the dispensing device.

The carrier is preferably made of a material that absorbs or retains water, such as filter paper, paper towel,
Examples include papers such as tissue paper, glass fiber filters, rayon, synthetic fibers such as polyester, cloth, nonwoven fabric, and cotton. In order to provide the carrier in the dispensing device, the material formed into a suitable size is usually packed from the side opposite to the discharge port of the dispensing device. In some cases, the carrier may be adhered to the dispensing device with an adhesive.

In order for the components of the reagent to be accommodated in the carrier, the carrier may be impregnated before being placed in the dispensing device, or may be impregnated after being placed in the dispensing device. Then, freeze-drying may be performed before or after placing in the dispensing device, or air-drying may be performed with a stationary device or a drier.

3. Liquids Liquids that may contain the object to be measured include:
Examples include biological samples (samples derived from humans, animals or plants), foods, beverages, and environmental samples.

Examples of the biological sample include blood, serum,
Biological fluids such as plasma, saliva, sweat, urine, feces, semen, vaginal secretions, tears, cerebrospinal fluid, ascites, amniotic fluid; liver, heart, brain, bone,
Extracts of organs, tissues, and cells such as hair, skin, nails, muscles, and nerve tissues; extracts or suspensions of feces; extracts of cells or cells; and extracts of plants. Foods include, for example, meat, seafood, vegetables, fruits, and processed foods.
Drinks include, for example, tap water, well water, and milk. Examples of the environmental sample include soil, air, seawater, river water, and lake water. Other liquids include, for example, water and organic solvents.

4. Substance in Liquid The substance in the liquid is not particularly limited as long as it is a substance that can react with the components of the reagent arranged in the dispensing instrument, and examples thereof include an object to be measured.

More specifically, amylase, lipase,
Acid phosphatase, prostatic acid phosphatase, alkaline phosphatase, creatine kinase, AST
(GOT), ALT (GPT), lactate dehydrogenase, leucine aminopeptidase, γ-glutamyl transpeptidase, cholinesterase, α-HBDH, N-acetylglucosaminidase and the like or isozymes thereof; total cholesterol, free cholesterol, HDL cholesterol, LDL cholesterol, neutral fat, β-
Lipoproteins, phospholipids, free fatty acids, oxidized LDL,
Lipids such as lipid peroxide; ammonia, urea nitrogen (U
N), nitrogen-containing components such as uric acid, creatinine and creatine; proteins such as total protein, albumin and globulin; amino acids such as tyrosine and histidine; glucose, hemoglobin A1c, fructosamine, glycoprotein and 1,5-anhydro-D-glucitol Bilirubin or bilirubin derivatives such as direct bilirubin, indirect bilirubin, unconjugated bilirubin, conjugated bilirubin, δ-bilirubin, conjugate C; vitamins such as vitamin A, vitamin C, vitamin E; lactic acid; Organic acids such as pyruvic acid; sodium ion, chloride ion, potassium ion, calcium ion,
Electrolyte components such as magnesium ion and inorganic phosphorus; iron, total iron binding ability, unsaturated iron binding ability, copper, zinc, lead, aluminum, lithium, mercury, chromium, manganese, cadmium,
HBs antigen, anti-HBs antibody, HBe antigen, anti-HBe antibody, anti-HBc antibody, HCV
Antigens, anti-HCV antibodies, anti-HIV antibodies, anti-ATLV antibodies and other virus-related antigens or antibodies; Escherichia coli O157 antigen;
Bacteria-related antigens or antibodies such as anti-Treponemal pallidum (TP) antibody, anti-mycoplasma antibody, anti-streptolidine O antibody (ASO); immunoglobulin G (IgG);
Immunoglobulin A (IgA), Immunoglobulin M (Ig
M), immunoglobulins such as immunoglobulin E (IgE); inflammatory markers such as C-reactive protein (CRP), α1-acid glycoprotein, haptoglobin, complement C3, complement C4, rheumatoid factor (RF); human villi Hormones such as sex gonadotropin (HCG), luteinizing hormone (LH), follicle-stimulating hormone (FSH), insulin, triiodothyronine (T3), serum total thyroxine (T4), growth hormone, glucagon; α-fetoprotein, polyamine , CEA, PA, tumor markers such as CA19-9; allergens, allergen-related antigens or antibodies such as allergen-specific IgE antibodies; antithrombin III (A
TIII) and other blood coagulation-related substances; fibrin body degradation products (FDP), D-dimer, protein S and other fibrinolytic-related substances; ABO blood group antibodies, blood group-related antigens and antibodies such as irregular antibodies; viruses; DNA or RNA of a bacterium; DNA or RNA of a bacterium; DNA or RNA of an animal or a plant such as a human; substances related to other diseases such as lipoprotein (a), ferritin, myoglobin; pesticides such as paraquat; toluene, trichloroethylene; 1,1,1-
Examples include organic compounds such as trichloroethane, sarin, and dioxin; toxic substances such as cyanide and azide compounds; and drugs such as theophylline, digoxin, and phenobarbital.

5. Procedure for using and dispensing the dispensing device of the present invention The procedure for the purpose of reacting a substance present in a liquid with a component of a reagent in the dispensing device is usually performed in the following manner.
It is as follows. A liquid is drawn up into the dispensing device of the present invention. The sucked-up liquid dissolves, suspends, or mixes the components of the reagent placed in the dispensing device. At this time, the reaction between the substance present in the liquid and the components of the reagent is performed in the dispensing instrument. Next, a fixed amount of the solution or suspension in which the reaction has been performed is discharged. The next step operation is performed on the fixed amount of the discharged solution or suspension.

In the present invention, the term “a fixed amount of the solution or suspension that has been discharged” refers to the components of the reagent placed in the dispensing device, the substances present in the sucked liquid, and / or And the like.

The procedure for dissolving, suspending, or mixing the components of the reagent arranged in the dispensing device with a liquid is usually as follows. A liquid is drawn up into the dispensing device of the present invention. The sucked-up liquid dissolves, suspends, or mixes the components of the reagent placed in the dispensing device. Following this, the following operation may be performed. Dispense a certain amount of this solution, suspension or mixture. The next operation is performed on the fixed amount of the discharged solution, suspension or mixed solution.

In the present invention, the "a fixed amount of the discharged solution, suspension or mixture" contains at least the components of the reagent arranged in the dispensing device. When the components of the reagent are in a liquid state, the components of the reagent arranged in the dispensing instrument may be discharged to a container, and the components of the reagent and the liquid may be mixed in the container.

6. Measurement method when used for measurement Examples of the measurement method when the present invention is used for measurement include biochemical reaction measurement, immunological reaction measurement (immunotrap method, immunoturbidimetry, latex nephelometry, indirect agglutination reaction, E
IA, FIA, RIA, LIA, immunochromatography),
Blood coagulation time measurement and nucleic acid probe method are mentioned. 7. Method of operation As a method of operation, any of a manual method and an automation method may be used.

8. Method for reacting a component in a liquid with a component of a reagent The method of the present invention for reacting a substance in a liquid with a component of a reagent includes the following steps: a) aspirating the liquid into the dispensing device in which the components of the reagent described above are arranged; b) dissolving, suspending or mixing the components of the reagent arranged in the dispensing device; c) thereby dissolving the components in the liquid. The substance is reacted with the components of the reagent in the dispensing device.

By "suctioning the liquid into the dispensing device on which the components of the reagent are placed", when the components of the reagent placed on the dispensing device are in a solid state, this is "dissolved" in the "liquid". Or, when the components of the reagent placed in the dispensing device are in a liquid state, they are "mixed" with the "liquid". Table 1 shows examples of biochemical reactions among the reactions between substances in the liquid and components of the reagent.

[0052]

[Table 1]

Table 2 shows examples of the immunological reaction.

[0054]

[Table 2]

Table 3 shows examples of the nucleic acid reaction.

[0056]

[Table 3]

Table 4 shows examples of the sugar-lectin reaction.

[0058]

[Table 4]

Table 5 shows examples of the ligand-receptor reaction.

[0060]

[Table 5]

Table 6 shows examples of other reactions.

[0062]

[Table 6]

The present invention has the following advantages. 1. Operation can be omitted for several steps. 2. The components of the reagent can efficiently participate in the reaction. (The minimum required amount can be involved in the reaction.) Cost can be reduced. (The amount of liquid and reagent to be used is the minimum required. There is no waste.) By separating the components of the reagent, it can be used for a long time.
(By separating the components of the reagent into those that are arranged in the dispensing device and those that are contained in the liquid, it can be stored for a long period of time.)

[0064]

EXAMPLES Example 1 Measurement of HBs Antigen in Serum Sample-1) (1) Preparation of Anti-HBs Antibody Immobilized Plate Acrylic resin extruded plate (25 mm wide, 60 mm deep, 1 mm thick) [Acrysandy Extruded acrylic resin (width 10 mm, depth 40 mm, thickness 0.
5 mm) [manufactured by Acrysandy Co., Ltd.] with a solvent at intervals of 5 mm, and grooves (width 5 mm) were formed on the surface shown in FIG.
mm, a depth of 40 mm and a height of 0.5 mm).

The plate shown in FIG. 1 surrounded by two extruded acrylic resin plates is 5 mm from the left end and 10 m from the left end.
m portion, anti-HBs antibody solution (Sinotest Co., Ltd., 5 μg
/ Ml dissolved in 10 mM phosphate buffer at a pH of 7) with a pipette and contacted,
The mixture was allowed to stand at 37 ° C. for 3 hours.

Next, the solution was removed by suction with a pipette, and 0.3 ml of 0.5% (w / v) casein-containing Tris buffer (pH 7.5, 50 mM) (hereinafter referred to as diluent A) was added. In addition, the mixture was allowed to stand at 4 ° C. overnight, and this was removed by suction with a pipette.

Then, an acrylic resin extruded plate (width 25 mm, depth 50 mm, thickness 1 m) is placed on the left end of the plate-like body.
m) [Acry Sunday] was attached with a solvent to cover the plate as shown in FIG. 1A. In this way, the specific binding substance (anti-H
A plate-like body having a band-shaped covering portion formed with Bs antibody) was prepared.

(2) Preparation of Anti-HBs Antibody-Fixed Particle Dispersion Particles (magnetic particles) [Dynabeads M-450 uncoated, manufactured by Dynal, particle size: 4.5 μm, coefficient of variation of particle size (C.I.
V. ) 5% or less, specific gravity 1.5, concentration 3% (w / v)] 1
ml and an anti-HBs antibody solution (manufactured by Sinotest, pH 7.0)
Was dissolved in 10 mM phosphate buffer at a concentration of 0.1 mg / ml) and reacted at 37 ° C. for 30 minutes. Here, about 20 times the amount of the diluent A was added to perform masking. Next, the obtained particles are washed with diluent A, and the particles are diluted with diluent A so that the concentration becomes about 0.4% (w / v).
Was redispersed. Thus, an anti-HBs antibody fixed particle dispersion was prepared.

(3) Placement of anti-HBs antibody-immobilized particles in a micropipette chip Micropipette (Eppendorf reference 4910)
(2) in a micropipette tip (volume: 100 μl; manufactured by Assist) using Eppendorf.
50 μl of the anti-HBs antibody-fixed particle dispersion prepared in the above was sucked in.

Liquid nitrogen was put in a polystyrene foam box, and the micropipette tip removed from the micropipette was immersed in the liquid nitrogen for 1 minute to disperse the anti-HBs antibody-fixed particle dispersion in the micropipette chip. Frozen immediately. Next, the micropipette chip in which the anti-HBs antibody-fixed particle dispersion was frozen was put into the freeze dryer chamber that had been cooled to −40 ° C. in advance, and the pressure was reduced to 0.1 Torr.

Then, while maintaining the degree of vacuum of 0.1 Torr, the temperature in the chamber is gradually increased to 28
Drying was performed at 10 ° C. for 10 hours. Thus, anti-H
Bs antibody-immobilized particles were placed in a micropipette tip.

(4) Measurement of HBs Antigen The sample HBs was measured using a micropipette (Eppendorf Reference 4910; manufactured by Eppendorf) in which a micropipette tip in which the anti-HBs antibody-immobilized particles prepared in (3) above were encapsulated was set. 100 μl of antigen-positive serum was aspirated. Thereby, the anti-HBs antibody-immobilized particles are dissolved and suspended in the sample, and the H
The reaction between the Bs antigen and the anti-HBs antibody immobilized on the particles was performed.

The anti-HBs antibody-immobilized particles are dissolved depending on the sample.
After confirming the suspension, the plate-shaped body was placed near the boundary between the covered portion and the uncovered portion of the groove, and was brought into contact with the strip-shaped covering portion.

Thereafter, a magnet is provided in the side surface direction of the plate-shaped body where the band-shaped covering portion is formed, so that the anti-HBs
Anti-H at the bottom of the groove of the plate-like body so that the antibody-immobilized particles move.
Anti-HB on the bottom of the groove from the part where Bs antibody is not fixed
The magnetic flux density is 40-60 in the direction in which the antibody is fixed.
A Gaussian magnetic field was generated. Within 3 minutes after the generation of the magnetic field, an image of particles as shown in FIG. 1C was observed.
When measurement was performed using negative serum as a sample, no particle image was observed (D in FIG. 1).

Example 2 (Measurement of HBs Antigen in Blood Sample-2) (1) Preparation of Anti-HBs Antibody-Immobilized Plate Body Immobilization of anti-HBs antibody as described in (1) of Example 1 A plate was produced. (2) Preparation of Anti-HBs Antibody-Fixed Particle Dispersion In the same manner as described in Example 1, (2), an anti-HBs antibody-fixed particle dispersion was prepared.

(3) Arrangement of anti-HBs antibody-immobilized particles in a micropipette chip In the same manner as described in (3) of Example 1, the anti-HBs antibody-immobilized particles were arranged in a micropipette chip. (4) Measurement of HBs antigen 100 μl of HBs antigen-positive blood was pipetted near the boundary between the covered and uncovered portions of the groove of the anti-HBs antibody-fixed plate prepared in (1) above. And placed in contact with the strip-shaped coating.

A tissue paper (trade name: Kimwipe, manufactured by Jujo Kinbari) is 3 cm × 3 cm × 4 mm
A sample absorber prepared by folding the sample into a (thickness) is brought into contact with the end of the plate-shaped body on which the anti-HBs antibody is immobilized, on which the band-shaped covering portion is formed, and the sample contacted with the band-shaped covering portion is absorbed. Removed.

Next, 100 μl of distilled water was applied by a micropipette (Eppendorf reference 4910; manufactured by Eppendorf) in which a micropipette tip on which the anti-HBs antibody-immobilized particles prepared in the above (3) were placed was set.
μl was aspirated. Thus, the anti-HBs antibody-immobilized particles were dissolved and suspended in distilled water. After confirming that the anti-HBs antibody-immobilized particles were dissolved and suspended in distilled water, the plate was placed near the boundary between the covered portion and the uncovered portion of the groove of the plate-shaped body, and was brought into contact with the band-shaped covered portion.

After that, the side of the plate-shaped body where the band-shaped covering portion is formed is turned downward, and the anti-H
Anti-HB on the bottom of the groove from the part where Bs antibody is not fixed
The anti-HBs antibody-fixed plate was inclined at an angle of about 60 ° so that the anti-HBs antibody-immobilized particles moved in the direction of the portion where the s antibody was immobilized. Fig. 1 within 3 minutes after tilting
The image of the particles as shown in C of FIG. When measurement was performed using negative blood as a sample, no image of particles was observed (D in FIG. 1).

[0080]

According to the present invention, the number of steps can be reduced as compared with the prior art when reacting a substance present in a liquid with a reagent component and when dissolving or suspending a reagent component in a liquid. In addition, the amounts of liquids and reagents to be used can be kept to a minimum, thereby enabling cost reduction.

[Brief description of the drawings]

FIG. 1 is a diagram for explaining Examples 1 and 2.

──────────────────────────────────────────────────続 き Continued on the front page (51) Int.Cl. ⁷ Identification symbol FI Theme coat ゛ (Reference) // G01N 33/48 G01N 33/53 T 33/53 35/06 K F term (Reference) 2G042 AA01 CB03 HA06 HA07 HA10 2G045 AA13 AA16 BA01 BB01 BB12 CA25 CA26 CB03 FB02 FB03 FB15 HA02 HA09 HA14 JA11 2G058 AA09 BA01 CC08 EA16 ED14 FA07 GA02 4G057 AB11 AB38 4G068 AA02 AB15 AE03 AE04

Claims (7)

[Claims] 1. A dispensing device for sucking up a liquid and discharging a fixed amount of the liquid, wherein a component of a reagent is arranged therein. 2. The dispensing device according to claim 1, wherein the reagent is a measurement reagent. 3. The dispensing device according to claim 1, wherein a reaction between a substance in the sucked-up liquid and a reagent component is performed in the dispensing device. 4. After the components of the reagent placed in the dispensing device are dissolved, suspended or mixed by the sucked liquid, or after reacting with the substance in the sucked liquid, a certain amount of the reagent is dispensed. The dispensing device according to any one of claims 1 to 3, which is discharged from the device. 5. The dispensing device according to claim 1, wherein the liquid is sucked up and the components of the reagent arranged in the dispensing device are dissolved, suspended or mixed, whereby the substance in the liquid and the reagent are mixed. A method of reacting a substance in a liquid with a component of a reagent, wherein the component is reacted in the dispensing device. 6. The method according to claim 5, wherein the reagent is a measurement reagent. 7. The method according to claim 5, wherein a certain amount of a reaction between a substance in a liquid and a reagent component in the dispensing device is discharged from the dispensing device.