JP3484180B2 - Reaction vessel - Google Patents

Description

Description: BACKGROUND OF THE INVENTION [0001] 1. Field of the Invention [0002] The present invention relates to a reaction vessel used for a method for measuring a biological substance. 2. Description of the Related Art The measurement of a substance related to a living body is carried out as a daily test in the field of environmental hygiene and the field of medicine, for example. In particular, in the medical field, many types of biologically related substances are measured in many facilities for the purpose of identifying a disease, determining a therapeutic effect on the disease, and the like. [0003] Such a measurement of a bio-related substance is performed by using a reaction with a specific affinity substance having a specific affinity for the bio-related substance. For example, acquired immunodeficiency syndrome (hereinafter referred to as AI), which has recently become a public health problem
Infectious diseases such as DS) and cancer-related substances that have been difficult to identify in the past can be measured by using antigen-antibody reaction, which is one of the reactions of biological substances and specific affinity substances. Has become. It is also possible to measure DNA or RNA, which is a gene of an infectious microorganism, using a complementary nucleic acid that binds to a characteristic portion of the nucleic acid. The reaction between the nucleic acid and the complementary nucleic acid is one of the reactions of the biologically relevant substance and the specific affinity substance. In addition, for example, a reaction of insulin, which is one of the hormones, and an insulin receptor is also one of the reactions of a biologically relevant substance and a specific affinity substance. [0004] As described above, many methods for measuring biological substances using a reaction with a specific affinity reaction are known.
In any of the measurement methods, the amount of a biologically-related substance (hereinafter, referred to as a substance to be bound) to which a specific affinity substance has bound must be measured. [0005] Such a method for measuring a biological substance includes the following:
The method is roughly divided into the following two methods. One is a homogeneous measurement method for determining a substance to be bound by utilizing the fact that the specific affinity substance and the substance to be bound bind to each other, thereby changing the properties of the specific affinity substance itself or the tracer bound thereto ( (Homogeneous method). The other is to make the complex of the specific affinity substance and the substance to be bound insoluble by any means, and then to separate the substance to be bound and the free substance B (B
This is a heterogeneous measurement method (heterogeneous method) that requires an operation of (round) / F (Free) separation. [0006] In the heterogeneous measurement method, a complex of a specific affinity substance and a substance to be bound is further combined with a second specific affinity substance that specifically binds to the complex, thereby converting the complex into a large molecule. There is known a method of making a complex insoluble by performing the method. However, this method is not preferable because it lacks reliability in measurement accuracy. Therefore, in recent years, a specific affinity substance is bound to an insoluble substance, and after reacting the specific affinity substance with the biological substance, a complex of the biological substance, the specific affinity substance, and the insoluble substance is converted from the sample solution. In general, B / F separation is performed by separation.
As such an insoluble substance, for example, a reaction tube, beads, filter paper and the like are used. However, in the above-mentioned heterogeneous measurement method, when the reaction tube itself is used as an insoluble substance, it is necessary to wash the reaction tube because it is used only once. However, there is an advantage that a relatively small amount of washing water is required, but there is a disadvantage that a large amount of waste is generated, which is not suitable for processing a large amount of a sample. Further, since it is considered that the reaction between the substantial bio-related substance and the specific affinity substance occurs on the surface of the reaction tube, there is a problem that the specimen and the reagent are unnecessarily consumed. On the other hand, when a substance other than the reaction tube, such as beads or filter paper, is used as the insoluble substance, the reaction tube can be used repeatedly. It becomes complicated and the device becomes large when the operation is automated. In addition, there is a problem that the insoluble substance is liable to carry over the sample (carry over). Furthermore, the accuracy and efficiency of the heterogeneous measurement method depends on how effectively the B / F separation is performed. On the other hand, when the above-mentioned reaction tube is used for an insoluble substance, the reaction tube and the sample solution can be separated relatively easily, but when an insoluble substance other than the reaction tube is used, for example, a filter is used. Insoluble substances are separated from a sample solution by using such a method, or insoluble substances are collected and separated by, for example, centrifugal force or magnetic force. However, it is difficult to accurately and quickly carry out the separation. As means for solving the drawbacks of the conventional non-uniform measurement method, the present applicant has proposed a sample introduction path having a cross-sectional area capable of aspirating a sample into a reaction vessel by capillary action, and a sample introduction path having the same. The sample is dropped into the sample introduction path by using a reaction container composed of a dent provided on the inner wall of the container and a transparent plate provided above the dent and having a flat surface that regulates the upper limit of the reaction part. A method has been proposed in which a sample is aspirated by a certain amount into a reaction part by capillary action to analyze a biologically relevant substance that forms a pattern of aggregated particles in a depression. This method is suitable for a trace measurement method in which measurement is performed on a small amount of a sample. But,
Since the reaction between the bio-related substance and the specific affinity substance generally requires a reaction time of several minutes to several tens of minutes, it is necessary to keep the sample solution in the reaction vessel for a certain period of time.
On the other hand, in the operation of washing the reaction vessel, it is not necessary to make the washing liquid stay in the reaction vessel as in the case of the sample solution. As described above, in the method for measuring a biological substance using a reaction container as described above, it is required to efficiently hold and remove the solution in the reaction container in order to make the operation more efficient. In order to achieve this demand, in a method for measuring a biological substance using a reaction container having the above-described structure, it is considered that a suction device is provided at one end of a sample introduction path to suck a cleaning liquid. However, in order to completely clean the reaction container, it is necessary to continuously supply the cleaning liquid for a certain period of time, and it is necessary to provide a cleaning liquid supply device capable of continuously supplying the cleaning liquid at the other end. As a result, the size of the reaction vessel itself can be reduced, but the size of the entire apparatus increases, and it is not possible to provide a method for measuring a biological substance using a small apparatus that can be used even in a small-scale laboratory. SUMMARY OF THE INVENTION The present invention has been made in view of the above circumstances, and provides a method for measuring a biological substance and a reaction container used therefor, which can remove a sample solution and a washing solution from the reaction container by simple means. The purpose is to: [0013] The reaction vessel of the present invention comprises a reaction vessel.
A reaction vessel main body and at least one reaction vessel main body.
One end also becomes an opening exposed on one end surface of the reaction vessel body
So that it is formed through the inside of the reaction vessel body,
A reaction section for holding a sample solution or a washing solution;
A transparent member provided on at least one of the side surfaces of the container body.
A light portion, provided on one end surface side of the reaction vessel body,
A water absorbing member capable of absorbing a sample solution or a washing solution is
A first position at which the aqueous member is separated from the opening;
The water absorbing member is moved from the first position by
Movably supported between the opening and a second position in contact with the opening
And a supporting member. According to the reaction vessel of the present invention, the supporting member
When holding the sample solution or the washing solution in the reaction section, the water absorbing member is separated from the solution. On the other hand, when removing the sample solution or the washing solution, the water absorbing member is brought into contact with the solution. Thus, the sample solution or the washing liquid is absorbed by the water absorbing member. Embodiments of the present invention will be described below with reference to the drawings. FIG. 1 is an explanatory view showing an example of a reaction vessel used in the method for measuring a biological substance according to the present invention. 11 in the figure
Is a substantially rectangular flat base material. On the main surface of the base material 11,
The two spacers 12 and 13 are arranged facing each other at an appropriate interval to form a groove 14. Further, a substantially U-shaped cutout 15 having a width larger than the width of the groove 14 is formed at a portion corresponding to one end of the groove 14 on the main surfaces of the spacers 12 and 13. A cover 17 in which a concave portion 16 having a width substantially equal to the width of the groove portion 14 is formed in a portion corresponding to the portion. Here, the base material 11, the spacers 12, 13 and the cover 17 are adhered to each other. The reaction container body 20 having the above-described structure is used.
Is a substrate 11, spacers 12, 13 and a cover 17
Reaction part 21 surrounded by a notch, notch part 15, base material 11 and injection part 22 defined by spacers 12 and 13
And a removal portion 23 defined in the concave portion 16 and the groove portion 14. In the reaction vessel body 20 as described above,
It is preferable that the thickness of the spacers 12 and 13 and the width of the groove 14 be set so that when the sample solution or the cleaning solution is injected into the injection section 22, the sample solution or the like is diffused into the reaction section 21 by capillary action. Specifically, spacer 1
Preferably, the thickness of the layers 2 and 13 is about 1 mm. The width of the groove 14 is preferably slightly larger than the tip of a commonly used pipette, for example, 3 to 5 mm. When the measurement of a biological substance is performed by measuring fluorescence or luminescence induced by the presence of the biological substance itself or a substance bound to the biological substance,
One of the base material 11 and the cover 17 needs to be made of a transparent material. In addition, a specific affinity substance is
1. When binding to the spacers 12, 13 or the cover 17, it is preferable to select a material according to the specific affinity substance. On the other hand, a support member 24 made of an elastic member is connected to the end of the reaction vessel main body 20 where the removed portion 23 is formed as follows. That is, the support member 24
Is a fitting groove 25 for fitting the reaction vessel body 20 to the inner edge.
Are formed. A substantially semicircular water-absorbing member 28 is provided at a position facing the end surface of the reaction container body 20. And the arm portions 26, 2
The reaction vessel main body 20 is fitted and adhered to the fitting groove 25 at the distal end of the reaction vessel 7. At this time, the water absorbing member 28 is set so as to be located at a predetermined distance from the removing section 23. Here, examples of the absorbent member 28 include absorbent cotton, pulp, cloth, sponge, paper, cellulose,
Carboxymethylcellulose, generally used in sanitary products and agriculture
Highly water-absorbing polymers (eg, grafted starches, polymerized polyacrylates, etc.) used as soil water retention agents in the horticultural field and the like can be used. The super-water-absorbing polymer is particularly preferable because the solution is hardly discharged even when pressure is applied from the outside. A method for measuring a biological substance using the reaction container 10 described above will be described. First, a specific affinity substance is disposed inside the reaction section 21. For example, the base material 11, the spacers 12, 1
A specific affinity substance is chemically or physically bound to at least one surface of the cover 3 or the cover 17 according to a conventional method. Further, the insoluble substance to which the specific affinity substance is bound may be arranged inside the reaction section 21 so as not to hinder the movement of the solution. Next, a sample solution in which a body fluid such as blood, urine, or cerebrospinal fluid is used as it is or subjected to an appropriate pretreatment is dropped into the injection section 22 with a pipette. The sample solution is supplied to the reaction section 21
Drop enough to fill. The dropped sample solution is quickly diffused into the reaction part 21 by a capillary phenomenon. In this manner, the sample solution is left inside the reaction section 21 for a predetermined reaction time to bind the biologically relevant substance in the sample solution to the specific affinity substance. At this time, if necessary, the reaction vessel 10 is heated to about 37 ° C.
It may be cooled to 0 ° C. or lower. Further, if the reaction unit 21 is left open for a long time, the sample solution evaporates and accurate measurement cannot be performed. Therefore, if necessary, the reaction container 10 may be stored under high humidity conditions. After the reaction, the outer surface of the support member 24 corresponding to the water-absorbent material 28 is pressed in the direction of the reaction vessel body 20 to deform the support member 24 and remove the water-absorbent material 28 from the removal portion 23. To contact the sample solution held in the reaction section 21. As a result, the sample solution is absorbed by the water absorbing material 28 and is removed from the reaction section 21. Thus, the water absorbing member 28 is pressed until the sample solution is completely removed from the reaction section 21. After removing the sample solution, in order to perform B / F separation, an appropriate amount of a washing liquid is dropped into the injection section 22 and the reaction section 21 is separated.
To spread. Thereafter, in the same manner as the sample solution described above,
The water absorbing material 28 is brought into contact with the cleaning liquid in the removing unit 23, and the cleaning liquid is absorbed by the water absorbing material 28 and removed from the reaction unit 21. This operation is repeated several times to completely separate the biologically relevant substance bound to the specific affinity substance from the free substance. Through the above operation, the biological substance can be bound to the reaction section 21 or the insoluble substance disposed inside the reaction section 21. Next, in order to measure the bound biological substance, for example, a substance that binds to the biological substance and induces a luminescence reaction (eg, luminol, lucigenin,
A binding substance (for example, antigen or antibody) holding peroxidase, glucose oxidase, etc.
2 and diffused into the reaction part 21. After reacting for a certain period of time, the binding substance is removed from the water-absorbing material 28 in the same manner as described above.
And removed from the reaction section 21. By the above operation, the bio-related substance, the binding substance, and the substance that induces the luminescence reaction are bound to the reaction section 21 or the insoluble substance. Next, a measurement of a biological substance is performed. First, an appropriate amount of a substance (for example, aqueous hydrogen peroxide, catalytic metals) that causes a luminescence reaction corresponding to the substance that induces a luminescence reaction is dropped into the injection unit 22 and diffused into the reaction unit 21. Here, firefly luciferin, a benzothiazole derivative, or the like may be added to enhance the luminescence reaction. After reacting for a certain period of time, the substance causing the luminescence reaction is absorbed by the water-absorbing material 28 and removed from the reaction section 21 in the same manner as described above. Thereafter, a detection unit of a measuring device such as a photomultiplier tube is positioned above the cover 17 covering the reaction unit 21, and the amount of light emitted from the reaction unit 21 is measured. At this time, it is preferable to perform complete light shielding in order to measure a relatively small amount of light emission. From the luminescence amount measured in this way, the amount of the biological substance in the sample solution is determined from a calibration curve created in advance. Here, instead of using the luminescence reaction, a fluorescent substance is used to measure the fluorescence emitted from the luminescent substance bound to the biological substance. it can. As described above, according to the method for measuring a biological substance of the present embodiment, the sample solution and the washing solution can be easily removed from the reaction section 21 in the reaction vessel 10 having a relatively simple structure. Further, without using a suction device or a continuous supply device of the cleaning liquid, etc., B
/ F separation can be performed. In the present embodiment, the reaction vessel provided with the support member made of an elastic member has been described as an example, but the configuration of the reaction vessel is not limited to this. Further, in the reaction vessel of the present embodiment, the reaction section is formed between two flat plates, but the structure of the reaction section is not limited to this. However, since the best effects are obtained when the present invention is applied to a reaction vessel having a reaction section capable of sucking or holding a predetermined amount of a sample solution or the like by capillary action as in the present embodiment, such a requirement is satisfied. A reaction vessel having a reaction section of any shape is preferred. In addition, the measurement of a biological substance is not limited to the method of measuring luminescence or fluorescence, and a method which can be generally used for the measurement of a biological substance can be adopted. further,
The above-described method for measuring a biological substance and the reaction container can be automated using an automatic analyzer to further improve work efficiency. As described above, according to the reaction vessel of the present invention, the water absorbing member is brought into contact with the sample solution or the washing liquid held in the reaction part by the support member, if necessary. These solutions can be efficiently removed with a simple structure. As a result, it is possible to reduce the labor required for the measurement operation of the bio-related substance, and to achieve effects such as easily achieving downsizing of the entire measurement device.

BRIEF DESCRIPTION OF THE DRAWINGS FIG. 1 is an explanatory view showing an example of a reaction vessel used in the method for measuring a biological substance of the present invention. [Description of Signs] 10: reaction vessel, 11: base material, 12, 13 ... spacer, 17 ... cover, 20 ... reaction vessel main body, 21 ... reaction section, 22 ... injection section, 23 ... removal section, 24 ... support member , 2
8 ... water absorbing member.

Claims (1)

(57) Claims 1. A reaction vessel main body, and at least one end provided in the reaction vessel main body has a reaction vessel.
The reaction vessel so as to form an opening exposed at one end face of the vessel main body.
A sample solution or
A reaction part for holding the cleaning liquid and at least one of the side parts of the reaction vessel main body.
A transparent portion, and the sample solution provided on one end surface side of the reaction vessel main body.
Alternatively, a water-absorbing member capable of absorbing a cleaning liquid is provided.
A first position where is separated from the opening, and the water absorbing member
Can be moved between the opening and a second position in contact with the opening.
And a supporting member for supporting the reaction container.