HK1095630B - Cartridge for automatic measurement and measuring device using the same - Google Patents

Description

Cartridge for automatic measurement and measuring apparatus using the same

Technical Field

The present invention relates to a cartridge for automatic assay for automatically determining a component contained in a sample by being incorporated in an automatic assay device, and to an automatic assay device for automatic assay using the cartridge.

Background

An analyzer for automatically analyzing human blood and the like, and a cartridge for automatic measurement by being provided in an analyzing apparatus have been proposed (JP 11-316226A). Such a cartridge includes a reaction well for performing a reaction, and a plurality of storage wells containing reagents used in the reaction. Further, in order to cope with the difference in dilution factor of the sample required according to the analysis items, a cartridge equipped with a dilution tank for diluting a predetermined amount of the sample to a required dilution factor has been proposed (international publication No. 01/84152).

When analysis is performed using the above-described automatic measuring apparatus, it may be necessary to subject a sample to pretreatment depending on the analysis items. For example, in the case of measuring a virus antigen in blood, it is generally necessary to (1) destroy virus particles, (2) expose and extract the virus antigen, (3) inactivate antibodies to the virus antigen contained in a sample, and the like, and for these purposes, heat treatment is required. The heat treatment is usually carried out at a temperature of 60 ℃ or higher. Therefore, in the conventional automatic measuring apparatus having only a temperature control section adapted to carry out an enzyme reaction at about 37 ℃, pretreatment at a high temperature cannot be carried out.

For the above reasons, in the present case, an operator is required to heat-treat a sample with another heating device (heating block or the like) in advance, and thereafter, it is required to supply the sample to an automatic measuring device, which takes a great deal of labor and time. Therefore, even when an automatic measuring apparatus is used, such a process cannot be considered to be completely automated. Further, even if a plurality of different analysis items to be analyzed can be measured simultaneously by an automatic measuring apparatus, it is necessary to separately handle the analysis items requiring heat treatment, which causes inconvenience.

DISCLOSURE OF THE INVENTION

An object of the present invention is to provide a cartridge for automatic measurement used in an automatic measuring apparatus, which is capable of automatically performing heat treatment on a sample, and a measuring apparatus using the cartridge.

The inventors of the present invention found that a heat treatment well for performing heat treatment of a sample can be provided in a cartridge, and the heat treatment well can be heated in an apparatus including the cartridge, so that analysis and measurement of the sample and heat treatment can be automated, thereby accomplishing the present invention.

The present invention provides:

(1) a cartridge for use in conducting an assay for a component to be measured contained in a sample, characterized by comprising: at least one heat treatment tank for performing heat treatment of the sample, and a reaction tank for reacting a component to be measured in the sample with a substance specifically reacting therewith.

Viewed from another aspect the invention provides:

(2) the cartridge according to (1), wherein the reaction between the component to be measured and the substance specifically reacting therewith is an immunological reaction;

(3) the cartridge according to (1) or (2), further comprising a reagent well for containing a reagent required for conducting an assay, a dispensing well for dispensing a sample, a diluting well for diluting a sample, a washing well for washing a reaction product, and/or an assay well for assaying a reaction product;

(4) the cartridge according to any one of (1) to (3), wherein the respective wells are linearly arranged, and the heat-treatment wells are located at either end of the cartridge;

(5) the cartridge according to any one of (1) to (4), wherein a well containing no solution or substance that is thermally affected is provided adjacent to the heat-treatment well;

(6) the cartridge according to any one of (1) to (5), wherein the heat-treatment well is separable from a well disposed adjacent thereto;

(7) the cartridge according to any one of (1) to (6), wherein a depth of the solution in a liquid form that affects a measurement value according to its capacity in the well is smaller than a depth of the diluting solution or the washing solution;

(8) the cartridge according to (7), wherein the solution which affects the measurement value according to the capacity thereof in the well is a solution containing a substance selected from the group consisting of a sample, a labeled antibody and a magnetic powder;

(9) the cartridge according to any one of (1) to (8), wherein information on information selected from the group consisting of sample information, analysis item information, reagent management information, and measurement calibration curve is recorded on the cartridge;

(10) the cartridge according to (9), wherein the information is recorded using a bar code;

(11) the cartridge according to (9) to (10), wherein the recording body of the information is provided so as to be breakable upon use of the cartridge, whereby it can be determined whether the cartridge has been used;

(12) the cartridge according to any one of (1) to (11), wherein a calibration mark showing the amount of a sample required for measurement is provided to the dispensing well;

(13) the cartridge according to any one of (1) to (12), wherein the cartridge is used by being incorporated in an assay device comprising a cartridge housing portion for housing the cartridge, a dispensing portion for dispensing a reagent and/or a sample into each well on the cartridge housed in the cartridge housing portion, an assay portion for assaying a reaction product on the cartridge housed in the cartridge housing portion, and a temperature control portion capable of controlling at least a heat-treatment well and a reaction well of the cartridge at a predetermined temperature;

(14) the cartridge according to (13), wherein the heat-treatment well and the reaction well can be controlled at desired temperatures different from each other when the cartridge is incorporated in the assay device;

(15) the cartridge according to (13) or (14), wherein a heat-treatment well is provided at one end of the cartridge, the heat-treatment well being positioned on a rear surface of the measuring device when the cartridge is set in the measuring device.

In addition, the present invention provides:

(16) an assay device comprising at least a cartridge housing section for housing the cartridge according to any one of (1) to (15), a dispensing section for dispensing a reagent and/or a sample into each well on the cartridge housed in the cartridge housing section, an assay section for assaying a reaction product on the cartridge housed in the cartridge housing section, and a temperature control section capable of controlling at least a heat-treatment well and a reaction well of the cartridge at desired temperatures different from each other;

(17) the measuring apparatus according to (16), wherein the temperature control section is capable of controlling the heat treatment bath at 50 ℃ to 100 ℃ and the reaction bath at 25 ℃ to 40 ℃; and

(18) the measuring apparatus according to (17), wherein the temperature control section is composed of two separately controlled heating blocks.

Brief description of the drawings

FIG. 1 shows a view of one embodiment of the cartridge of the present invention. Part a represents a top view and part B represents a cross-sectional view.

FIG. 2 is a view showing another embodiment of the cartridge of the present invention. Part a represents a top view and part B represents a cross-sectional view.

FIG. 3 is a view showing an example of movement of a solution when an assay is carried out using the cartridge of the present invention.

FIG. 4 is a view showing another example of movement of a solution when an assay is carried out using the cartridge of the present invention.

Best mode for carrying out the invention

Unless otherwise specified, terms used herein have the following meanings.

The cartridge of the present invention can be used when a component to be measured contained in a sample is measured, and is generally set in an automatic measuring device for use.

The above measurement is usually carried out by a method (hereinafter, this is generally referred to as "the measurement method of the present invention") for measuring a component to be measured in a sample, which comprises the steps of dispensing a sample containing the component to be measured, reacting the component to be measured contained in the sample with a substance specifically reacting therewith, and measuring the amount of the reaction product. The assay method of the present invention typically further comprises the step of performing a heat treatment of the sample. In addition, the measurement method of the present invention preferably further comprises a step of diluting the sample.

The component to be measured (which may be referred to as "analysis item" hereinafter) is not particularly limited, and any component may be used as long as a substance specifically reacting with the component is present. Examples of the combination of the component to be measured and the substance specifically reacting therewith include: antigens and antibodies, antibodies and antigens, enzymes and substrates, and sugar chains and lectins. Therefore, in the present invention, the expression "specifically occurring reaction or specific reaction" refers to a specific biochemical binding. The component to be measured or the substance specifically reacting therewith may be a substance whose chemical property changes before and after the binding, such as a substrate. Above all, the cartridge of the present invention is particularly suitable for the measurement of a component to be measured which requires heat treatment of a sample. One example of such a component to be tested includes a viral antigen, and a specific example thereof includes an HCV core antigen and a chlamydia antigen.

The sample may be any sample as long as it contains the above-mentioned component to be measured, or may contain the component. Examples include blood, serum, plasma, and urine. These samples may be heat-treated in a heat-treatment well, if necessary, depending on the selected analysis items. The presence/absence of heat treatment, heating temperature and heating time, and the like can be appropriately set according to the selected analysis item. For example, in the case of the above-mentioned HCV core antigen, the heat treatment may be carried out at, for example, 65 ℃ for 30 minutes. Further steps of suitable cooling of the sample (which also includes cooling by air-holding the sample) may also be required where the heat-treated sample is considered to have a high temperature and may negatively affect the cell in which the sample is provided. However, this cooling step is not necessary in the case where the temperature of the sample does not cause any problem due to the mixing of the sample with a large amount of solution, for example, in the case where the sample is diluted using a dilution tank (described later).

The conditions of the step of reacting the component to be measured with a substance specifically reacting therewith and the step of measuring the amount of the reaction product and the like may be appropriately selected depending on the combination of the component to be measured with a substance specifically reacting therewith. For example, the reaction of the enzyme with the substrate and the measurement of the amount of the reaction product can be realized by mixing the enzyme with the substrate to allow the enzyme to act on the substrate and measuring the amount of the reaction product (degradation product of the substrate). The reaction of the antibody and the antigen and the determination of the amount of the reaction product can be performed by mixing the antibody or the antigen, a solid phase carrier to which the corresponding antigen or antibody can be bound, and a label to form a reaction product (immunocomplex), washing the reaction product to remove unreacted antibody or antigen and unreacted label from the immunocomplex (B/F separation), and determining the amount of the label bound to the solid phase by forming the immunocomplex. Therefore, in the present invention, the expression "determining the amount of the reaction product" includes not only directly determining the amount of the reaction product itself but also determining the amount of a substance quantitatively related to the amount of the reaction product. From the thus-determined amount of the reaction product, the amount of the component to be measured in the sample can be calculated.

The cartridge of the present invention is characterized by comprising at least a heat-treatment well for performing heat treatment of a sample, and a reaction well for reacting a component to be measured in the sample with a substance specifically reacting therewith. With this configuration, even in the case where an analysis item requiring heat treatment of a sample is selected, all steps including heat treatment can be automatically performed.

In a preferred embodiment of the cartridge of the present invention, the respective slots are linearly aligned. In this case, it is preferable to place the heat-treatment well at either end of the cartridge. Further, a well which does not contain a solution or a substance which is affected by heating is preferably placed adjacent to the heat treatment well. Such a well that does not contain a solution or substance that is affected by heating may be a well that is not used (there may be a well that is not used depending on the case of an analysis item because the cartridge of the present invention may be applicable to many kinds of items to be analyzed). Further, the heat treatment bath may be spaced apart from the bath positioned adjacent thereto.

By providing the cartridge as described above, when the heat-treatment well is heated, the influence of the heating on the reagent and the reaction can be reduced or prevented.

The cartridge of the present invention is generally used by being set in an assay device including a cartridge housing portion for housing the cartridge, a dispensing portion for dispensing a reagent and/or a sample to each well on the cartridge housed in the cartridge housing portion, an assay portion for assaying a reaction product on the cartridge housed in the cartridge housing portion, and a temperature control portion capable of controlling at least a heat-treatment well and a reaction well of the cartridge at a desired temperature. In a preferred embodiment of the cartridge of the present invention, when the cartridge is set in the measuring device, each of the heat-treatment well and the reaction well is controlled by a temperature control section of the measuring device. Generally, the heat treatment bath and the reaction bath are controlled at different temperatures. However, in the case where the reaction temperature is high, the heat treatment bath and the reaction bath may be controlled at the same temperature. Specifically, for example, the heat treatment bath may be controlled at 50 ℃ to 100 ℃, preferably 60 ℃ to 70 ℃, and the reaction bath may be controlled at 25 ℃ to 40 ℃, preferably 33 ℃ to 38 ℃. These temperatures may be appropriately set according to the items to be analyzed. For example, the temperature of the heat treatment bath may be determined according to the presence/absence of heat treatment, heating temperature, heating time, and the like of the item to be analyzed. In addition, the temperature of the reaction tank can be determined, for example, according to the conditions in the above-described reaction step and the step of measuring the amount of the reaction product. The reaction vessel is preferably positioned apart from the heat treatment vessel so that it is not thermally affected when the heat treatment vessel is heated.

Further, when the cartridge is set in the measuring apparatus, it is preferable that the heat treatment well is provided at an end portion of the back surface of the measuring apparatus. Specifically, in this embodiment, the cartridge of the present invention has a proximal end and a distal end, the distal end is located on the rear surface of the measuring device as compared with the proximal end when the cartridge is set in the measuring device, and the heat-treatment well is located on the distal end of the cartridge. This makes it difficult for a person using the cartridge of the present invention to reach the heat-treatment well.

The cartridge of the present invention may further have a dilution well for diluting a predetermined amount of the sample to a desired dilution factor. For the measurement, depending on, for example, the kind, concentration, measurement principle, etc. of the sample or component to be measured, a certain analytical method such as an enzyme immunoassay (EIA method), a latex immunoassay (LIA method), a turbidimetric immunoassay (TIA method), a fluorescence immunoassay (FIA method), or a chemiluminescent enzyme immunoassay (CLEIA method) may be selected. Further, depending on the above conditions, the sample may be assayed in an undiluted liquid form, or the sample may need to be assayed after dilution. In addition, depending on the conditions, the dilution factor of the sample may be changed. Even in this case, if a dilution well for diluting a predetermined amount of sample to a desired factor is provided, the operator dispenses only a predetermined amount of sample regardless of the analysis item in the measurement operation, thereby reducing the labor required for the operator to confirm the dispensing amount and greatly reducing the possibility of measurement failure due to errors in the dispensing amount. In addition, in the automatic measuring device provided with the cartridge of the present invention, a mechanism for changing the amount of sample dispensed according to the analysis item is not required, so that the device can be simplified. In the present invention, in the case where the diluting solution is not filled in the diluting well, when the diluting step is performed in the same manner as the dilution is required in the analysis item, it is apparent that the sample remains as a non-diluted liquid (that is, the dilution factor is 1). Thus, the term "diluting" in this specification includes maintaining the sample as an undiluted liquid. Further, in the case of diluting the sample with a high factor, it is preferable that at least two dilution wells are provided in the cartridge, and the dilution is performed by at least two stages. The dilution factor of the sample and the dilution solution to be filled in the dilution tank may be appropriately selected depending on the kind of the sample, the component to be measured, a substance specifically used for the reaction with the component to be measured, and the like. The dilution solution may contain reagents required for sample pretreatment, and in this case, dilution and pretreatment may be performed simultaneously in the dilution tank. The pretreatment includes treatment with an acid, an alkali, an organic solvent, a protein denaturant, an enzyme, and a protease inhibitor, treatment with a surfactant, and the like. The tank containing the pretreatment liquid may be provided separately from the dilution tank.

In addition, the cartridge of the present invention has a reagent well for containing a reagent necessary for measurement of a component to be measured contained in a sample. The reagent tank may also be used as a reaction tank. In other words, some of the reagents participating in the reaction may be contained in the reaction tank. The reagent contained in the reagent tank or the reaction tank may include one or more kinds as long as the contained reagents do not react with each other. The reagent to be contained may be a liquid (e.g., a solution or a suspension) or a solid as long as it can be dissolved or suspended in a solution to be injected into the tank.

Further, the cartridge of the present invention preferably further comprises a dispensing well for dispensing a sample (hereinafter, may be referred to as "sample well"). Thus, a predetermined amount of sample may be added from the dispensing well to the dilution well into which the sample has been previously dispensed by a consistent process. Further, when dispensing a sample from a container in which the sample has been collected in the cartridge, it is not necessary to perform strict quantity control, which makes the operation by the operator easier. Further, in the automatic measuring device in which the cartridge of the present invention is to be disposed, it is not necessary to provide an additional device such as a mechanism for directly quantifying and dispensing a sample from a main sample container other than the cartridge in order to dispense a predetermined amount of the sample to the cartridge, thereby simplifying the device. Further, if a calibration mark showing the amount of a sample required for measurement is provided on the dispensing well, a required amount of a sample can be easily and accurately dispensed to the cartridge. The distribution tank may also be used as a heat treatment tank.

A wall may be provided around the distribution chute. This avoids problems such as leakage of the sample dispensed to the dispensing well into contact with the operator's hand, sample flow into another well, sample adhesion to the aluminum seal sealing the well, and contamination of the pipette tip used to pierce the aluminum seal. The wall to be provided may be disposed around the periphery of the groove. For example, in the case where the dispensing well is provided on the side (proximal end) of the handle portion closest to the one end of the cartridge, a straight wall may also be provided between the handle portion and the dispensing well, and between the dispensing well and another adjacent well. The wall may also be provided separately or may protrude continuously from the inner wall of the groove. Since the possibility of leakage of the dispensed sample can be reduced, it is preferable that the wall be formed to continuously protrude from the inner wall of the well. The wall may have a dimension of about 0.5mm to 5mm, for example. FIG. 2 shows an example of a cartridge having such a distribution groove. In the cartridge, a sample well (1), a labeled antibody well (2), a washing well (5), (3), a washing well (4), a washing well (3), (5), a labeled antibody reaction well (6), a magnetic powder well (7), a washing well (1), (8), a washing well (2), (9), a photometric well (10), a diluting solution well (1), (11), a diluting solution well (2), (12), a luminescent substrate well (13), a diluting well (1), (14), a diluting well (2), (15), and a heat treatment well (16) are arranged linearly in this order. Furthermore, the sample well (1) is provided with the wall.

The cartridge of the present invention may have a measuring well for measuring the amount of the reaction product. For example, a photometric cell for optical measurements can be provided. Here, for example, in the case where a special measurement condition is required (for example, in the case where measurement under dark conditions is required), the measurement well may be provided in another cartridge, or may be provided as a separate measurement well. For example, only the measuring well is made of a material capable of blocking light and combined with the cartridge, thereby eliminating the need for facilities such as a dark room.

The shape and size of the cartridge of the present invention are not particularly limited. However, for the convenience of the operator's hand, the cartridge preferably has a shape such as a boat in which reagent wells, sample distribution wells, dilution wells, reaction wells and/or measurement wells are linearly arranged. Multiple slots may be used for each type of slot. Further, in order to perform measurement of a plurality of analysis items to be analyzed, two or more lines of well groups necessary for measurement of a component to be measured may be arranged in parallel. The material of the cartridge of the present invention is not particularly limited. However, in the case of storing a reagent or the like hermetically, a material having stability not reacting therewith may be selected. In addition, in the case of performing heat treatment at high temperature, a material excellent in heat resistance and heat conductivity can be selected.

In the cartridge of the present invention, the reaction between the component to be measured and the substance specifically reacting therewith is preferably an immunological reaction. That is, the component to be measured and the substance specifically reacting therewith are preferably an antibody and an antigen.

As the immunoreaction, a method of simultaneously reacting a sample, a first substance specifically immunoreactive with a component to be measured in the sample, and a second substance specifically immunoreactive with a first immunocomplex generated by the reaction (hereinafter referred to as "one-step method"), and a method of reacting the above substances in two stages (hereinafter referred to as "two-step method") are known.

In the present invention, it is preferable to use a two-step process. Specifically, for example, a component to be measured in a sample interacts with a substance specifically immunoreactive therewith to form a first immunocomplex, and then the first immunocomplex reacts with a label specifically immunoreactive therewith to form a second immunocomplex. In this case, the cartridge of the present invention preferably has a reaction well for forming the first immunocomplex and a second reaction well for forming the immunocomplex. More specifically, the cartridge of the present invention has washing wells for B/F separation corresponding to the respective reaction wells. These washing wells may be filled with a washing solution in advance, or may be dispensed from another cartridge or bottle for filling, for example.

For such an immune reaction, a method of B/F separation using a magnetic force in a liquid suction/discharge line provided in a dispensing portion of the device employs a first substance immobilized on magnetic particles that can be collected by the magnetic force. Specifically, for example, a method (e.g., patent No.3115501) of performing B/F separation by bringing a magnet into contact with a pipette tip, a hose, a stainless steel tube, or other flow line serving as a suction/discharge liquid from a test cassette well from the outside and collecting magnetic particles on the inner wall surface thereof, or the like can be used.

The reagent and/or solution for use in the present invention, which is required for the measurement of the component to be measured contained in the sample, may be previously filled in another cartridge, which may be used in combination with the cartridge of the present invention at the time of measurement. For example, the assay can be carried out by filling in advance a diluting solution of the sample, a substance and a label specifically reacting with the component to be measured in the sample, a washing solution for washing the immunocomplex in another cartridge, and the like in another cartridge, and distributing the reagent and/or the solution to each well on the cartridge of the present invention by a unified operation. By this means, the mechanism of the apparatus can be simplified, and the structure of the cartridge of the present invention can be simplified and reduced. In addition, the problem of storage stability of the reagent and/or solution to be used can be easily solved. Of course, it is also possible to fill both the cartridge of the present invention and another cartridge with reagents and/or solutions required for the assay and use the two cartridges in combination.

All reagents and/or solutions necessary for the determination of the component to be measured in the sample can be filled in the cartridge of the present invention. All necessary reagents such as a diluting solution of the sample, a substance and a label specifically reacting with the component to be measured in the sample, and a washing solution for washing the immunocomplex, etc. may be filled in the cartridge of the present invention in advance. Thus, all procedures for one component to be measured can be handled in one cartridge, which can reduce the waste of reagents. The supply or discharge of water becomes unnecessary, so that the measuring apparatus can be further simplified and the time required for the measurement can be reduced.

In another preferred embodiment of the present invention, the depth of the solution in liquid form that affects the measurement value according to its capacity in the tank may be set to be smaller than the depth of the diluting solution or the washing solution. Here, the term "depth" refers to the distance from the bottom surface of the tank to the surface of the solution. In the apparatus in which the cartridge of the present invention is to be disposed, a dispensing portion in which a pipette tip or the like can be mounted is provided, and a liquid is injected into or aspirated from a well of the cartridge through the pipette tip. At this time, when the depth of the solution affecting the measurement value according to its capacity in the well is large, the solution will adhere to the outside of the pipette tip and be taken into another well, which makes it impossible to perform accurate measurement. The inventors of the present invention have noted this problem and have proposed the following ideas: by setting the depth of the solution to be smaller than that of the diluting solution or the washing solution, even if the solution is brought into another well, the solution adhering to the pipette tip can be removed with the diluting solution or the washing solution when the pipette tip is inserted into the well containing the diluting solution or the washing solution. Further, it is preferable that the depth of the tank containing the solution affecting the measurement value according to the capacity thereof is set to be small according to the depth of the solution. With this structure, the loss of the trace amount of the solution adhering to the cell wall or the like can be reduced, and the measurement can be performed with high accuracy. Examples of the solution that affects the measurement value according to its capacity in the well include a solution containing a sample, a labeled antibody, a magnetic particle, and the like.

When the cartridge of the present invention is previously filled with solutions such as reagents and/or diluting solutions, labels, washing solutions, etc., it is preferable that the top surface of the cartridge is sealed with aluminum foil, plastic film, etc. to prevent contamination of foreign substances and evaporation/deterioration of reagents. The sealing of the aluminum foil is particularly preferred because such sealing is easily opened automatically by a piercing mechanism in an automatic measuring device or by the leading end of a pipette tip or the like. In the case where a reagent and/or a solution or the like is filled in another cartridge and measurement is performed in conjunction with the cartridge, the cartridge is preferably sealed as well.

In another embodiment of the cartridge of the present invention, information, such as sample information, analysis item information, reagent management information, and information of a calibration curve for measurement, is recorded on the cartridge. For example, such information may be recorded by printing or attaching a barcode on the cartridge. Further, a magnetic recording device, an IC chip into which information is input, or the like may be connected. For example, by providing a barcode or the like to the cartridge, if an automatic measuring device that can recognize the barcode of the cartridge and automatically select an analysis item is used, the operator only needs to select the cartridge, and thus an arbitrary analysis item can be easily and efficiently measured by using one automatic measuring device. Further, it is possible to easily measure a plurality of analysis items to be analyzed without fail without operating a work chart, which is a main cause of errors in setting of analysis items by a conventional ordinary automatic measuring apparatus. In addition, storage and management of the reagent are also easy. Further, even if the operator does not input the information on the calibration curve into the automatic measuring device, the information on the calibration curve can be automatically input into the device. The record of information, such as a bar code, may be arranged to be broken when the cartridge is used, so that it can be determined whether the cartridge has been used. For example, when information is recorded on a sealing member for sealing the well, the sealing member may be broken during use by a punching mechanism of the automatic measuring apparatus, a tip of a pipette tip, or the like, and thus the recording body may be broken. For this reason, whether or not the cartridge has been used can be automatically determined.

In the measurement method of the present invention, in the case where a sample contains a plurality of components to be measured, it is preferable to simultaneously measure the plurality of components to be measured by using a plurality of cartridges or cartridges in which two or more lines of cell groups are provided in parallel. In this case, it is preferable to use an automatic measuring apparatus capable of measuring a plurality of analysis items to be measured simultaneously in parallel and in which a plurality of the cartridge of the present invention having wells corresponding to the plurality of analysis items to be measured (two or more rows of well groups are provided in parallel) can be set, or an automatic measuring apparatus in which the cartridge of the present invention can be set.

In the automatic apparatus in which the cartridge of the present invention is set in use, there may be used respective known mechanisms such as a mechanism for sucking a predetermined amount of liquid from one well and dispensing it into another well, a mechanism for mixing the contents in the wells, a mechanism for performing B/F separation, a mechanism for measuring the amount of a reaction product or a labeled substance, a mechanism for calculating the amount of a component to be measured from the measurement result of the amount of the reaction product or the labeled substance, a mechanism for controlling the temperature of the cartridge, a mechanism for recognizing a barcode, a mechanism for simultaneously performing measurements of a plurality of cartridges, and the like.

The invention will be described below with reference to an immunoassay, more particularly to an example of a chemiluminescent enzyme immunoassay (CLEIA), as an example of a preferred aspect.

The cartridge according to a preferred aspect is a cartridge for automatic measurement by being set in an automatic measuring device which automatically measures the amount of a component to be measured in a sample. This cartridge has: a reaction tank for reacting the component to be measured with a substance specifically immunoreactive therewith; a plurality of reagent tanks for filling reagents, each to be used in a reaction; a sample distribution groove for distributing the sample; a dilution tank for diluting a sample; a heat treatment bath for heating the sample; a washing tank for performing B/F separation; and/or a photometric cell. As described above, the reagent tank may also be used as a reaction tank. These slots are preferably used as described below. A sufficient amount of the diluting solution sufficient to dilute a predetermined amount of the sample to a desired dilution is filled into the diluting well. The plurality of reagent tanks are filled with a solid-phase carrier for specifically performing an immunoreaction, a labeled antigen or antibody, a reagent for performing measurement of the amount of a label, and the like, respectively. The washing solution for washing the immunocomplexes is filled in the washing tank.

In addition, in the reagent well of the cartridge, for example, a solid phase carrier (sensitized solid phase) to which an antigen or an antibody is bound can be placed, and thus the well can also be used as a reaction well. The solid phase carrier may include polystyrene beads, magnetic particles, etc., which are conventionally used for immunoassays. Alternatively, instead of adding the solid-phase carrier to the tank, an antibody or an antigen may be used by attaching the antigen or the antibody to the inner wall of the tank.

The immunoassay used in this example is preferably a chemiluminescence enzyme immunoassay (CLEIA) method which is superior in sensitivity. The solid phase carrier is preferably a magnetic powder, by which B/F separation necessary for CLEIA can be easily performed by a magnet. The B/F separation can also be performed by applying a magnetic field to the cartridge from the outside of the cartridge by using a permanent magnet, an electromagnet, or the like. In addition, according to the method disclosed in Japanese patent JP11-262678A, a magnetic field can be applied by using magnets provided on the aspirating side and the dispensing side of a pipette tip or the like of the dispenser.

Other reagent wells may be used as the reaction well by adding a labeled antigen or antibody thereto. Examples of the labeling substance include, for example, enzymes, radioisotopes, coloring substances, fluorescent and luminescent substances, and various colored particles. In chemiluminescent enzyme immunoassays (CLEIA), it is preferred to use a plurality of enzymes. Examples of such labeling enzymes include alkaline phosphatase, peroxidase, galactosidase, and glucose oxidase. As the substrate for labeling the enzyme, those corresponding to the respective enzymes can be suitably used. For example, adamantyl methoxyphenyl-phosphoryl dioxane (AMPPD) can be used for alkaline phosphatase, luminol/peroxide can be used for peroxidase, and adamantyl methoxyphenyl- β -D-galactosyl dioxane (AMPGD) can be used for galactosidase.

When a dilution tank is used, a predetermined amount of a diluting solution required for each analysis item is preferably injected into the dilution tank in advance. For example, in the case of measuring two different analysis items, i.e., Hepatitis C Virus (HCV) antibody and hepatitis B surface antigen (HBsAg), the measurement of the above two analysis items can be simultaneously performed in the same analysis step by using two cartridges having the same amount of sample, the same amount of reagent solution of solid-phase carrier, the same amount of reagent solution of labeled antigen or antibody, the same amount of washing solution, the same amount of measurement conditions of label, and the like for the two analysis items, but having different amounts of dilution solution filled in a dilution tank for the above two analysis items, in an automatic measuring apparatus provided with two or more mechanisms capable of performing a series of immunoreaction processes in parallel.

In the case where the sample is to be highly diluted, it is preferable to provide two or more dilution wells on the cartridge so as to perform the dilution in two or more steps. Fig. 1 and 2 show examples of such a cartridge.

Further, as described above, the cartridge of the present invention may be used in combination with another cartridge containing a reagent and/or a solution required for the assay component to perform an assay task. For example, the cartridge of the present invention having a sample distribution well, a dilution well, a reaction well, a washing well and a photometric well can be used without filling any reagent and/or solution in each of the above-mentioned wells. On the contrary, in another cartridge, a diluting solution, a solid-phase carrier, a labeled antigen or antibody, a reagent for detecting the amount of the label, and the like are contained, and these substances are dispensed from the cartridge into the cartridge of the present invention, so that the measurement can be similarly performed.

In the sample dilution step, the sample may be pretreated by adding an acid, an alkali, an organic solvent, a protein denaturant, a detergent, or the like to the dilution solution. For example, in the case of using blood (whole blood) as a sample, since blood contains a large amount of interferon and for other reasons, it is preferable to add any desired detergent or the like for pretreatment. By simultaneously diluting and pretreating the sample in this manner, highly accurate detection can be conveniently performed even when blood is used as the sample. Therefore, doctors and nurses can preferably use the present invention in emergency trials and when care testing (POCT) is to be performed.

When the washing solution is supplied from a part of the apparatus in the automatic measuring apparatus as seen in the conventional automatic measuring apparatus, it takes much time and labor to prepare the washing solution for washing the unreacted sample and the label from the immunocomplex (B/F separation), and to constantly replenish the washing solution and treat the waste liquid during the measurement. In addition, the washing solution used in the conventional automatic measuring apparatus is standardized in composition and amount of liquid regardless of the items to be analyzed, so that it is impossible to use the washing solution having the optimum composition for each item to be analyzed. From the above viewpoint, it is preferable that the cartridge also contains a washing solution. However, in the case where the components or the amount of the washing solution are the same, for example, the washing solution is uniformly supplied by a part of the devices in the automatic measuring apparatus as described above.

For example, in the chemiluminescence enzyme immunoassay, the measurement of the label can be performed by mixing an immunocomplex and a substrate for a labeling enzyme, and then directly measuring the label in a photometric well using a photomultiplier tube or the like. For example, in the case where an assay well composed of a material capable of blocking light is used in combination with a cartridge, the assay can be performed by bringing one end of a photomultiplier tube into direct contact with the upper portion of the well, whereby the immunoassay can be conveniently performed in the absence of facilities such as a dark room. Further, in the case of an enzyme immunoassay, the immunocomplex is mixed with an enzyme substrate solution, and then measurement light having a measurement wavelength is irradiated from the bottom or side of the measurement well to measure light propagating through the measurement well.

The automatic measuring instrument of the present invention comprises at least a cartridge housing portion for housing a cartridge, a dispensing portion for dispensing a reagent and/or a sample into each well on the cartridge contained in the cartridge housing portion, a measuring portion for measuring a reaction product on the cartridge contained in the cartridge housing portion, and a temperature control portion for controlling at least a heat-treatment well and a reaction well in the cartridge at predetermined temperatures different from each other.

The above-mentioned cartridge bearing portion may be the same as a conventional cartridge bearing portion except that the cartridge bearing portion is made to have a structure capable of housing the cartridge of the present invention. The dispensing portion is constituted by a conventional mechanism such as a liquid drawing/dispensing mechanism or the like corresponding to the kind and properties of the reagent and/or the sample. The term "assign" as used herein is meant to include: transferring reagents and/or samples from one well to another well on the cartridge, and transferring reagents and/or samples from outside the cartridge to a well on the cartridge. The measuring part is constituted by a conventional mechanism such as a photometric mechanism, depending on the kind and properties of the reaction product. In the case of performing an assay using the cartridge of the present invention having two or more rows of parallel well groups or using a plurality of cartridges of the present invention, the automatic assay device is preferably one in which: having a plurality of mechanisms disposed in parallel therein for performing a series of immune reactions and capable of simultaneously operating and controlling, for example, sample dispensing, sample dilution, reagent dispensing, B/F separation, and photometric processes. Thus, even in the case of immunoassay, it is possible to simultaneously measure a plurality of analysis items using an instrument capable of performing only one analysis process without significantly increasing the man-hours required for measurement even when different analysis items are measured.

The portion of the dispensing portion that contacts the reagent and/or sample (pipette tip, etc.) is preferably replaceable. By replacing this portion for each test, contamination of the cartridge used in the subsequent measurement can be easily prevented. Further, as described above, it is preferable to perform B/F separation using a magnetic force in a pipette tip or the like.

The temperature control section controls at least the heat-treatment bath and the reaction bath at predetermined temperatures different from each other. Specifically, for example, the temperature control section controls the heat treatment bath at 50 ℃ to 100 ℃ and the reaction bath at 25 ℃ to 40 ℃. In a preferred embodiment, the temperature control section is composed of two separately controlled heating blocks.

Further, as described above, it is preferable that a barcode or the like is provided on the cartridge of the present invention, and measurement is performed by using a measuring instrument having a mechanism for recognizing the barcode. By using a detector that can recognize a barcode and can automatically select an analysis item, it is not necessary to individually set, for example, whether or not heat treatment, reaction temperature, and photometric conditions should be performed; and the assay results can be easily analyzed; the operator does not have to enter information about the calibration curve, etc. Thus, automatic measurement can be performed more easily and efficiently. More specifically, all measurement steps can be automatically performed only by the operator selecting a cartridge corresponding to a desired analysis item, so that measurement can be accurately performed with reduced human error.

Examples of the invention

The invention is explained in more detail below by way of examples. However, the following examples are merely illustrative, and the scope of the present invention should not be considered to be limited by the following examples. It will be apparent to those skilled in the art that any changes, improvements or modifications may be made without departing from the spirit of the invention.

< example 1> Cartridge for two-step reaction

FIG. 1 shows an example of a cartridge of the present invention. In this cartridge, a sample well (1), a labeled antibody well (2), a washing well (5), (3), a washing well (4), a washing well (3), (5), a labeled antibody reaction well (6), a magnetic powder well (7), a washing well (1), (8), a washing well (2), (9), a photometric well (10), a diluting solution well (1), (11), a diluting solution well (2), (12), a luminescent substrate well (13), a diluting well (1), (14), a diluting well (2), (15), and a heat treatment well (16) are arranged linearly in the stated order. This cartridge is sealed with an aluminum foil at the upper portion, and analytical items are recorded with a bar code on the aluminum foil.

In the case of performing measurement using the above-described cartridge and using an automatic measuring apparatus provided with a 6-pack type suction/discharge mechanism and a magnetic particle separation mechanism, the operation in this case will be described below (fig. 3). In this example, after the sample reacts with the magnetic particles to which the antibody specifically reacting with the component to be measured is bound, the reactant and the labeled antibody react with each other (two-step method). Further, this example includes a step of heat-treating the sample. In FIG. 3, the arrows represent the movement of the solutions, and the reference numerals on the arrows represent the order of the movement of the solutions.

1. The sample is dispensed into the sample well (1).

2. The reagent cartridge to which the sample is dispensed is loaded into the automatic measuring apparatus. At most six reagent test cartridges can be loaded at the same time. These cartridges can be used to measure a plurality of items to be analyzed which are different from each other and arranged in an arbitrary manner.

3. The automatic measuring device is started.

4. The automatic measuring machine reads the barcode attached to the reagent cartridge and recognizes the selected analysis item.

5. The diluted solution is sucked from the diluted solution tank 2(12), and the whole is discharged to the diluted solution tank 1 (14). Further, the diluted solution is sucked from the diluted solution tank 1(11), and the whole is discharged to the diluted solution tank 2 (15).

6. The diluting solution is aspirated from the diluting solution reservoir 2(12), and then the sample is aspirated from the sample reservoir (1). All of them are discharged to the dilution tank 2(15) to be mixed, thereby completing the first dilution step.

7. The sample diluted in the first step is aspirated from the dilution wells 2(15), and the whole is discharged to the dilution wells 1(14) to be mixed, thereby completing the second dilution step.

8. The sample diluted in the second step is aspirated from the dilution well 1(14) and discharged in its entirety into the heat-treatment well (16), thereby being heat-treated at a desired temperature for a predetermined time.

9. The sample is sucked from the heat treatment bath (16) and discharged into the magnetic powder bath (7) to be mixed with the magnetic powder, thereby allowing the sample to react with the magnetic powder for a predetermined period of time.

10. After the reaction, the sample is aspirated from the magnetic particle well (7), and the magnetic particles are separated by a permanent magnet in the pipette tip from which the sample has been aspirated (the magnetic particles are adsorbed on the inner wall of the pipette tip, and the solution is discharged), and moved to the washing well 1 (8). After washing the magnetic particles, the magnetic particles are separated by a permanent magnet and moved to the washing bath 2 (9). The magnetic particles were similarly washed again, and the magnetic particles were separated using a permanent magnet.

11. A labeled antibody solution is aspirated from the labeled antibody well (2) in a state where the magnetic particles are adsorbed on the inner wall of the pipette tip, and discharged together with the magnetic particles into the labeled antibody reaction well (6) to be mixed with each other, so that they react with each other for a predetermined period of time.

12. After the reaction, the sample is aspirated from the labeled antibody reaction well (6), and the magnetic particles are separated by a permanent magnet in the pipette tip from which the sample has been aspirated, and moved to the washing well 3 (5). After washing, the magnetic particles are further separated by a permanent magnet and moved to a washing tank 4 (4). Further, after washing, the magnetic particles are separated by a permanent magnet and moved to the washing tank 5 (3). The magnetic powder was washed again and separated with a permanent magnet.

13. A luminescent substrate is aspirated from a luminescent substrate well (13) in a state where the magnetic particles are adsorbed on the inner wall of the pipette tip, and discharged into a photometric well (10) together with the magnetic particles to be mixed with a luminescent substrate solution. After a reaction for a predetermined period of time has elapsed, the amount of luminescence is measured from the upper portion of the photometric well (10) by means of a photomultiplier tube (PMT).

< example 2>

Another example of performing an assay using the cartridge described in example 1 will be shown below (FIG. 4). In this example, the measurement was performed using the same apparatus as in example 1 except that the sample, the magnetic powder and the labeled antibody were simultaneously reacted with each other (one-step method). In FIG. 4, arrows represent the movement of the solution, and reference numerals on the arrows represent the order of the movement of the solution.

1. The sample is dispensed into the sample well (1).

2. The reagent cartridge to which the sample has been dispensed is loaded into the automatic measuring apparatus. At most six reagent test cartridges can be loaded at the same time. These cartridges can be used to assay a plurality of items to be analyzed which are different from each other and arranged in an arbitrary manner.

3. The automatic measuring device is started.

4. The automatic measuring instrument reads a bar code attached to the reagent measuring cassette and recognizes the selected analysis item.

5. The diluted solution is sucked from the diluted solution tank 2(12), and the whole is discharged to the diluted solution tank 1 (14). Further, the diluted solution is sucked from the diluted solution tank 1(11), and the whole is discharged to the diluted solution tank 2 (15).

6. The diluting solution is aspirated from the diluting solution reservoir 2(12), and then the sample is aspirated from the sample reservoir (1). All of them are discharged to the dilution tank 2(15) to be mixed, thereby completing the first dilution step.

7. The sample is aspirated from the dilution wells 2(15), and the whole is discharged to the dilution wells 1(14) to be mixed, thereby completing the second dilution step.

8. The sample is aspirated from the dilution well 1(14) and discharged in its entirety into the heat-treatment well (16), thereby performing heat treatment at a desired temperature for a predetermined time.

9. The sample is aspirated from the heat-treatment well (16), and then the labeled antibody solution is aspirated from the labeled antibody well (2), and discharged into the magnetic powder well (7) to be mixed with magnetic powder, thereby reacting them for a predetermined time.

10. After the reaction, the sample is aspirated from the magnetic particle well (7), and the magnetic particles are separated by a permanent magnet in the pipette tip from which the sample has been aspirated (the magnetic particles are adsorbed on the inner wall of the pipette tip, and the solution is discharged), and moved to the washing well 3 (5). After washing the magnetic particles, the magnetic particles are separated again by the permanent magnet and moved to the washing tank 4 (4). The magnetic powder is similarly washed, separated by a permanent magnet, and moved to a washing tank 5 (3). The magnetic powder is washed again and separated with a permanent magnet.

11. A luminescent substrate is aspirated from a luminescent substrate well (13) in a state where the magnetic particles are adsorbed on the inner wall of the pipette tip, and discharged into a photometric well (10) together with the magnetic particles to be mixed with a luminescent substrate solution. After reacting for a predetermined time, the amount of luminescence is measured from the upper part of the photometric well (10) using a photomultiplier tube (PMT).

Industrial applicability

According to the present invention, a measuring process including heat treatment of a sample can be automatically performed using an automatic measuring apparatus and a cartridge for automatic measurement used for the automatic measuring apparatus. Further, in a preferred embodiment of the cartridge of the present invention, the sample may be heated without affecting the reaction.

Claims (10)

1. A cartridge for automatic assay used by being set in an automatic assay device which automatically performs assay of a component to be assayed in a sample and which includes a dispensing portion for dispensing a reagent and/or a sample, comprising:

at least one heat treatment bath for performing heat treatment of the sample;

a reaction tank for reacting a component to be measured in the sample with a substance specifically reacting therewith;

a dispensing well configured for dispensing a sample;

a labeled antibody tank containing a labeled antibody solution;

a diluting solution tank containing a diluting solution for diluting a sample; and

a washing tank containing a washing solution for washing the reaction product;

the depth of the labeled antibody tank is smaller than that of the diluted solution tank or the washing tank;

the depth of the labeled antibody solution is less than the depth of the dilution solution or wash solution;

a well selected from among a labeled antibody well, a diluting solution well and a washing well is sealed with a sealing member; and is

Information selected from the group consisting of sample information, analysis item information, reagent management information, and calibration curve-related information for measurement is recorded on the sealing member, and the information recording body is configured to be disconnected upon use of the cartridge.

2. The cartridge according to claim 1, wherein the depth of the distribution well is smaller than that of the diluting solution well or the washing well.

3. The cartridge according to claim 1 or 2, wherein the reaction between the component to be measured and the substance specifically reacting therewith is an immune reaction.

4. The cartridge according to claim 1 or 2, further comprising an assay well configured to assay a reaction product.

5. The cartridge according to claim 1 or 2, wherein said respective wells are linearly arranged, and said heat-treatment well is located at either end of said cartridge.

6. The cartridge according to claim 1 or 2, wherein a well not containing a solution or a substance that is thermally affected is provided adjacent to the heat treatment well.

7. The cartridge according to claim 1 or 2, wherein the heat-treatment well is separated from a well disposed adjacent thereto.

8. The cartridge according to claim 1 or 2, wherein the information is recorded with a barcode.

9. The cartridge according to claim 1 or 2, wherein a calibration mark showing the amount of a sample required for measurement is provided to the dispensing well.

10. The cartridge according to claim 1 or 2, wherein the sealing member is an aluminum foil or a plastic film.