JP2001318101A - Cartridge - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a cartridge which eliminates a need or introducing a cleaning liquid from the outside when a measurement by an apparatus is completely automated by a method wherein a reagent which is required for immunoassay is sealed in one cartridge. SOLUTION: The cartridge comprises a reaction tank in which an immune agglutination reaction is performed. The cartridge comprises a plurality of storage tanks into which all reagents used for the immune agglutination reaction are filled independently. In the cartridge, the cleaning liquid is sealed into the storage tanks. The cleaning liquid cleans the inner wall of a pipette tip.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a cartridge incorporated in an automatic immunoassay apparatus for automatically quantifying a trace component present in a sample.

[0002]

2. Description of the Related Art Various specific selective reactions have conventionally been used in the field of so-called diagnostic agents which are useful for diagnosing and treating diseases by quantifying trace components in body fluids such as human blood. Have been. Such a specific selective reaction is intended to measure the amount of a trace component such as a disease marker present in a body fluid such as blood as a sample, and includes, for example, an antigen-antibody reaction. Can be
In addition, by utilizing such an antigen-antibody reaction,
So-called immunoassays and the like are performed.

[0003] Among such immunoassays, an assay using an immunoagglutination reaction is widely used as a simple operation. This measurement method is based on an immunoturbidimetric method in which a substance (such as an antibody or an antigen) having specific selective reactivity with a measurement target in a sample and the measurement target directly form an aggregate, A substance having specific selective reactivity with respect to a substance is supported on the surface of a carrier such as latex fine particles, and then the liquid containing the fine particles is brought into contact with a sample liquid to cause a specific selective reaction. There is a latex agglutination method in which an agglomerate is generated at least, and in each case, the absorbance that changes with the agglomerate is measured.

[0004] In recent years, improvements have been made in this field, and at present, automatic measuring devices for automatically performing all of the steps of the immunoassay have become active.

[0005] Japanese Patent Application Laid-Open No. 63-281053 discloses an automatic immunoassay by a solid phase method which is performed using a container having a plurality of wells such as a reaction well and a sample well having a fibrous matrix. However, in this method, in addition to the cartridge to be used, a device for supplying a reagent necessary for immunoassay is essential, and it is necessary to change the supplied reagent in accordance with the change of the substance to be measured. There is a problem that the configuration of the device becomes complicated.

[0006] JP-A-4-218775 discloses an automatic immunoassay using a container having a plurality of openings, at least one of which has a solid phase. However, this technology is essentially related to the technology in which the solid phase is magnetic particles, and has the disadvantage of poor versatility,
The operability was not always good.

[0007] Japanese Patent Application Laid-Open No. 11-316226 discloses a reaction tank for performing a reaction and a plurality of storage tanks filled with reagents, which are used for an enzyme-labeled immunoassay or a radioimmunoassay. There is disclosed a cartridge containing a carrier and filling all necessary reagents in a solution state in a storage tank. However, although this cartridge packs all the reagents, only the washing liquid is supplied from the outside, and the operability is not always good.

[0008] All of the conventional methods are immunoassays using a container having a plurality of wells, and are aimed at automating the measurement. However, these methods
Since only a part of the reagents necessary for the measurement are contained in the container or not at all, it is necessary to prepare the reagents necessary for the measurement separately from the container. There were problems such as complexity.

[0009]

In view of the above, the present invention has been made in view of the above, and provides a cartridge which can be easily incorporated into an automatic measuring apparatus and which can perform measurement simply by a complete one-cartridge system for one sample. The purpose is to do so.

[0010]

SUMMARY OF THE INVENTION The present invention has a reaction tank for performing an immune reaction and a plurality of storage tanks for filling reagents used for the immune reaction, and automatically removes trace components present in a sample. A cartridge used by being incorporated in an automatic measurement device for quantitatively determining the amount of an immunological reaction, wherein the automatic measurement is performed using an immunological substance having specific reactivity with a trace component in the sample; and Wherein the reaction tank has the immunological substance therein, and the plurality of storage tanks, inside thereof, independently, A cartridge characterized by being filled with all reagents necessary for an immune reaction. Hereinafter, the present invention will be described in detail.

[0011]

DESCRIPTION OF THE PREFERRED EMBODIMENTS The present invention relates to a cartridge used in an automatic measuring device for automatically quantifying trace components present in a sample. The shape of the entire cartridge is not particularly limited as long as it has a reaction tank and a plurality of storage tanks. For example, a boat-type one in which the reaction tank and the plurality of storage tanks are arranged in parallel is provided. (See FIG. 1).

Although the size of the cartridge is not particularly limited, it is preferable that the cartridge be smaller in order to be incorporated in an automatic measuring apparatus.
m, a height of about 0.5 to 5 cm, and a thickness of about 0.3 to 3 cm.

The size and shape of the reaction tank and the plurality of storage tanks are not particularly limited as long as they are well-shaped ones capable of storing a reaction and a reagent as described later. And a rectangular parallelepiped and the like. These can be configured to pierce the cartridge. The shape of the well itself may be cylindrical or prismatic, and the bottom surface may be circular or square. In the present invention, in order to optically observe a specific reaction progressing in the well from outside the well, it is preferable that a cartridge portion corresponding to at least a part of a side surface of the well is formed of a light-transmitting substance.

The well formed of the above-mentioned light-transmitting substance corresponds to a photometric cell in optical measurement, but may also serve as a plurality of storage tanks filled with a reagent before use.

The reaction tank and the storage tank of the cartridge are
In order to avoid contamination by foreign matter, it is preferable that the upper part is sealed.For example, an aluminum foil or a polymer film is applied to the upper surface of the cartridge with a hot melt adhesive, for example. What adhere | attached etc. are mentioned. Sealing with an aluminum foil is preferable because it can be easily broken by a seal breaker or the like provided in an automatic measuring device and has good sealing properties.

The material of the cartridge is not particularly limited, except that it is not a substance that inhibits a specific reaction carried out in a reaction vessel.
Inexpensive and easy-to-mold materials such as polyethylene resin and polypropylene resin are exemplified. For example, a transparent polystyrene resin can directly measure the absorbance with a spectrophotometer.

Further, a waste tank for disposing of a liquid (waste liquid) generated when a series of reactions and measurements are completed can be provided. By providing such a waste tank, it is possible to easily clean up the measurement work. The waste tank may be a diversion tank that is emptied during the measurement operation among the plurality of storage tanks, or may be separately provided.

The cartridge of the present invention may have a mechanism for docking a cuvette containing a sample liquid. Usually, a cuvette container containing a sample liquid which has been subjected to pretreatment such as centrifugation is set separately from the cartridge in the automatic measuring device. Then, the measuring device collects the sample liquid from the sample container and supplies it to the reaction in the cartridge. By having the mechanism as in the present invention, the space in the automatic measuring device can be effectively used, and miniaturization of the device can be expected.

The sample includes, for example, body fluids such as human blood. Trace components in a sample can be used for the diagnosis and treatment of human diseases by quantitatively measuring immunologically, for example, hepatitis virus,
Disease markers such as rheumatoid factor, C-reactive protein, hemolytic streptococcal toxin, and various enzymes are included.

In the cartridge of the present invention, an agglutinate is generated by exciting an immune reaction inside the cartridge, and a trace component in the sample is measured by reading an optical change with respect to the agglutinate. .

The immunological substance is not particularly limited as long as it has specific immunoreactivity with the above-mentioned trace components, and examples thereof include the above-mentioned trace component antibodies and the like. May be a monoclonal antibody or a polyclonal antibody, or may be a decomposed product of the antibody.

As the immunological substance, a case where a carrier is not required and a case where the immunological substance is carried on a carrier and subjected to a reaction can be appropriately selected. When using a carrier, in the present invention,
The carrier is present inside the reaction vessel. The carrier is
It is preferably a solid, more preferably fine-particle beads, and still more preferably polystyrene latex beads (hereinafter, the carrier supporting the immunological substance may be referred to as a sensitizing carrier. There is also). When an immunological substance is carried on a carrier, the carriers are aggregated by performing a specific reaction based on the immunoagglutination method.

The size of the latex fine particles serving as a carrier is preferably, for example, latex fine particles having an average particle size of 0.01 to 5 μm obtained by graft copolymerizing polyethylene with polystyrene, which is commonly used in the immunoagglutination method, and more preferably. Has a diameter of 0.1 to 0.7 μm.

The method for supporting (sensitizing) the above-mentioned immunological substance on the above-mentioned latex fine particles is not particularly limited, and for example, a conventionally known physical adsorption method or chemical bonding method can be used.

[0025] The plurality of storage tanks of the cartridge of the present invention include:
It is independently filled with all of the necessary reagents for immunoturbidimetry in solution. The `` independently '' means that each of the plurality of storage tanks may be filled with a separate reagent one by one, or one storage tank may be filled with a plurality of reagents. This means that there may be an empty storage tank. The “solution” also includes that some reagents are packed as a lyophilized product, and when used, they are made into a solution with a buffer solution or the like which is also filled.

Thus, the plurality of storage tanks are filled with all of the reagents necessary for the specific selective reaction and the reaction with the labeling substance. The gist of the present invention lies in that the reaction can be completed.

The use of the cartridge in this manner eliminates waste of the reagent and eliminates the need to prepare the reagent every time the measurement object is changed. That is, the cartridge to be used may be determined at the same time as the determination of the substance to be measured. For example, cartridges for each substance to be measured are independently prepared and applied, such as a cartridge for C-reactive protein and a cartridge for rheumatoid factor. As a result, there is no measurement error due to reagent confusion.

Of course, it is possible to simultaneously measure a plurality of items by independently enclosing, for example, reagents for measuring C-reactive protein and rheumatoid factor in each of a plurality of storage tanks in one cartridge. It is. In that case, the arithmetic circuit on the automatic measuring device side may be set to be freely written so that the measurement sequence can be freely changed.

A sample liquid is introduced into the reaction tank of the cartridge of the present invention by an appropriate method. For example, a cuvette may be set in the measuring device after a sample is directly introduced into the cartridge of the present invention by a technique, or a cuvette containing a sample solution may be set in the device side. A cuvette containing a sample liquid may be docked to the cartridge of the present invention. The sample liquid is human blood or the like as described above.

The trace component present in the sample solution causes an immune reaction in the reaction vessel by coming into contact with the immunological substance or a carrier (latex beads) carrying the immunological substance. The generation of an immune reaction initiates, for example, aggregation between beads and can be subjected to optical measurement.

In order for the above-mentioned immune reaction to proceed favorably, it is preferable to introduce a buffer solution for suspending latex beads and the like into the above-mentioned reaction tank in addition to the above-mentioned sample solution.
In the present invention, the buffer is filled in the storage tank as a reagent necessary for the reaction. The buffer is not particularly limited, and a buffer usually used for an antigen-antibody reaction or the like can be used. Examples thereof include a phosphate buffer containing 0.1% bovine serum albumin.

When a whole blood sample is used as a sample in addition to the above-mentioned suspension buffer, a buffer containing a hemolytic agent for destroying blood cell components (to be distinguished from a latex suspension buffer, the following dilution is used): (Sometimes referred to as a liquid) may be filled in the storage tank. As the type of hemolytic agent, saponin can be preferably used.

When a whole blood sample is used as a sample, there is a high possibility that an error occurs in the measurement result due to hemoglobin contained in red blood cells. Therefore, prepare a storage tank to be filled with a reagent for quantitative measurement of hemoglobin, calculate the hemoglobin concentration simultaneously with the sample measurement, calculate the hematocrit value using the obtained hemoglobin concentration, and correct the sample measurement value You can also.

After the specific selective reaction is completed and the optical measurement is completed, the residual mixture of the sample solution and the buffer solution is removed as a waste liquid out of the reaction tank by, for example, an aspirator. The waste liquid is discarded in the waste tank as described above.

In the present invention, the reagents (plural types) to be filled in the storage tank are each in the form of a solution. Due to the solution state, introduction from the storage tank to the reaction tank can be easily performed by a single operation using a pipette.

A pipette for sucking and discharging a solution is
In order to prevent contamination, it is preferable that the inside of the pipette is washed with a washing liquid or the like. The cleaning liquid can be an embodiment stored in any place of the cartridge of the present invention, which is a feature of the present invention.

The work for washing unreacted components in the immunoassay, that is, the washing solution required for the so-called B / F separation is usually large and varies depending on the kind of the trace component to be measured. Since the substance is unnecessary, it is usually supplied from some of the automatic measuring apparatuses without being contained in the cartridge.

However, the cartridge of the present invention is for an immunoagglutination reaction, and does not require B / F separation. Therefore, by eliminating the need for a large amount of cleaning solution and specializing only in the cleaning solution for pipette cleaning, the cleaning solution was also successfully sealed in the cartridge.

The washing solution is not particularly limited as long as it does not affect the immunoagglutination reaction, and examples include physiological saline and deionized water.

In the present invention, when each of the above-mentioned liquid reagents of the latex suspension, the diluting solution for hemolysis and the washing solution is put into the storage tank, it is preferable to fill each of them into another storage tank.

The number of the storage tanks can be appropriately determined depending on the number of reagents in solution, operability and simplicity.
It is 2 to 10, more preferably 3 to 8. FIG. 2 shows an example of filling each storage tank. As shown in the figure, an empty storage tank can be prepared and used for diluting a sample to be measured or a place to put a sample to be measured.

[0042]

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS The present invention will be described below in detail with reference to the drawings. FIG. 2 is an example showing the configuration of the cartridge of the present invention. A cartridge used in carrying out the present invention has cells (A, B, C) and wells (1 to 6). The cell is a reaction tank for performing photometry, and the well means a storage tank.

In addition to these cell wells, there is a tank containing a disposable pipette tip for performing pipetting before the cartridge is used, and a tank that becomes a waste tank after use.

Further, the apparatus has a sample tank used when a user directly injects a sample into the cartridge, and a cuvette for setting a sample in a general-purpose small tube or the like to the cartridge together with the tube. The change between the sample tank attached to the cartridge and the sample tube is optional, and can be easily achieved by instructing the apparatus to change the sequence.

Before use of the cartridge, the sample tank, well 1, cell ABC and waste tank are empty. Well 2 contains a hemoglobin measurement reagent for performing hemoglobin correction.
Well 3 contains a diluent for hemolysis (saponin-containing saline)
The well 4 is filled with a buffer, the well 5 is filled with a chip washing solution (distilled water), and the well 6 is filled with a latex suspension. Since the type of reagent changes depending on the item, the positional relationship between the extra cells and wells changes.

When whole blood, which is a sample, is injected into the sample tank of the cartridge and the cartridge is set in the apparatus, the following measurement procedure is automatically performed.

After the laminate seal of the cartridge is broken by the sharp tip of the pipette nozzle serving also as a seal breaker in the apparatus, the pipette nozzle on the apparatus side and the pipette tip contained in the cartridge are docked.

Next, a physiological saline 95 μl × 2 times from the well 3 to the well 1 by pipetting, a total of 190 μl
Is dispensed.

Next, the buffer solution was transferred from the well 4 to the cell B by 84.
Dispense μl.

Next, the hemoglobin measuring reagent is dispensed from well 2 to cell A twice at 77 μl × 2 times in a total of 154 μl.

Next, the physiological saline remaining in the well 3 was used
After the suction and discharge of 0 μl are performed twice, the chips are washed by transferring 50 μl of the distilled water in the well 5 to the waste tank.

Next, 10 μl of the sample in the sample tank is dispensed into the well 1, and suction and discharge with 110 μl are performed five times, followed by stirring, mixing and dilution.

110 μl of physiological saline remaining in well 3
Is performed twice, the distilled water 50 in the well 5 is removed.
The chip is washed by transferring μl to the waste tank.

Next, 28 μl of the diluted sample in well 1 was dispensed into cell A, and suction and discharge with 110 μl were performed five times.
Stir and mix. At this time, a change in absorbance is detected, and the hemoglobin concentration is measured. A hematocrit value is calculated from the obtained hemoglobin concentration.

On the other hand, the buffer remaining in well 4
After the suction and discharge of l are performed twice, the distilled water
The chip is washed by transferring 0 μl to the waste tank.

Next, 48 μl of the diluted sample in the well 1 is dispensed into the cell B, and suction and discharge with 85 μl are performed five times, followed by stirring and mixing. 110 μl of distilled water remaining in well 5
After the suction and discharge of the water are performed twice, the distilled water 11
The chip is washed by transferring 0 μl to the waste tank.

Next, the latex solution 28.2 μm in the well 6 was used.
1 is dispensed into the cell B, and suction and discharge with 110 μl are performed three times, followed by stirring and mixing. At this time, a change in absorbance is detected, and a target component concentration is measured.

Finally, the component concentration is corrected from the obtained component concentration and the hematocrit value obtained at the same time.

By automating each operation described above by the apparatus side, it is possible to carry out an automatic measurement method for continuously measuring a sample. The method of automating the pipetting operation by the apparatus and the configuration of the apparatus can be freely changed according to the purpose of measurement, the shape of the cartridge, and the like, and there is no particular limitation.

[0060]

According to the present invention, the reagents necessary for the measurement, including the cleaning solution, are enclosed in one cartridge, so that the measurement can be easily automated by the device, and the Thus, one cartridge can cope with all of the reagents, thus eliminating waste of reagents, making reagent management extremely simple and easy, enabling simple and quick measurement, and greatly contributing to the field of clinical testing.

[Brief description of the drawings]

FIG. 1 is a perspective view showing an embodiment of a cartridge of the present invention.

FIG. 2 is a sectional view showing an embodiment of the cartridge of the present invention.

 ──────────────────────────────────────────────────続 き Continued on the front page (72) Inventor Hiromasa Ota 57 Kyoto Nishi-Akita-cho, Minami-ku, Kyoto City, Kyoto

Claims (9)

[Claims] A reaction vessel for performing an immune reaction, and a plurality of storage tanks for filling reagents used for the immune reaction;
A cartridge used by being incorporated into an automatic measurement device for automatically quantifying a trace component present in a sample, wherein the automatic measurement uses an immunological substance having specific reactivity with the trace component in the sample. The immune reaction to be performed, and optical measurement of the aggregate based on the immune reaction, wherein the reaction tank has the immunological substance therein, the plurality of storage tanks, A cartridge characterized by being independently filled with all of the reagents necessary for the immune reaction. 2. The cartridge according to claim 1, wherein all necessary reagents are filled in a solution. 3. The cartridge according to claim 1, wherein a part of a necessary reagent is filled with a freeze-dried product and returned to a solution state when used. 4. The cartridge according to claim 1, wherein the plurality of storage tanks also serve as photometric cells for optical measurement. 5. The cartridge according to claim 1, further comprising a disposal tank for disposing of the waste liquid after the reaction. 6. The cartridge according to claim 5, wherein the waste tank diverts a used storage tank. 7. The cartridge according to claim 1, wherein a cuvette containing a sample can be docked. 8. The cartridge according to claim 1, wherein said immunological substance is carried on a carrier surface. 9. The cartridge according to claim 8, wherein the carrier is latex fine particles having a diameter of 0.1 to 0.7 μm.