JPH0815259A - Antigen or antibody detecting apparatus, detecting method, and kit for detection - Google Patents

Abstract

PURPOSE:To provide an antigen or antibody detecting apparatus, a detecting method and a kit for the detection which can detect an antigen or an antibody easily and quickly with excellent producibility and for which no specific consideration for storage and transportation is needed. CONSTITUTION:Regarding an apparatus to detect an antigen or an antibody by separating coagulated masses formed by coagulation by an antigen-antibody reaction from a sample and determining either the coagulated reaction product or the not coagulated reaction product, the apparatus is characteristically provided with a reaction container 10 in which a sample is to be stored and a formed member 12 produced by sintering to separate coagulated masses from the sample.

Description

Detailed Description of the Invention

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to an antigen or antibody detection device, a detection method and a detection kit using a sintered molded member.

[0002]

2. Description of the Related Art Recently, a gel centrifuge method has been attracting attention as a new test method which is excellent in simplicity and speed in blood tests such as blood group and antiglobulin. In the gel centrifugation method, for example, in the case of blood group test, centrifugation is performed by causing an antigen-antibody reaction in a tube containing the gel, and if aggregation occurs due to the antigen-antibody reaction, the aggregate cannot pass through the gel. The presence or absence of agglutination and its strength can be objectively determined by confirming that erythrocytes stop at the upper part and, if agglutination does not occur, pass through the gel and fall into the bottom of the microtube as clumps.

JP-A-1-163662 discloses the use of inert porous particles such as polyacrylamide gel particles to form a carrier-bound antibody-antigen complex or carrier-bound antigen-antibody complex in an aqueous medium. Methods and test kits for detecting antigens and / or antibodies of the type that optically visualize the complex have been described. Further, JP-A-4-285858 describes a method and an apparatus for confirming the presence of a blood group antigen or an antibody against the same, using a matrix composed of essentially incompressible microparticles. . The matrix used herein can be generated by, for example, centrifugal force, magnetic force, electric force, hydrostatic pressure, negative or positive pressure force, or long-term storage under normal gravity,
It is a non-porous particle that resists changes in shape or size.

[0004]

Whether the inert porous particles or non-porous particles disclosed in the above-mentioned prior art have a region (aggregation detection region) for trapping an agglutination reaction product by an antigen-antibody reaction. Not fixed. Therefore, when it is stored for a long period of time or shakes during transportation, or when the container filled with particles falls, the shape of the container flows to change its shape. In that case, it will take time and effort to put it back.
In addition, the gap between particles becomes non-uniform, which may deteriorate the reproducibility.

The present invention has been made in view of the above, and requires no special consideration for storage or transportation, obtains good reproducibility, and is a simple and rapid detection apparatus, detection method, and detection method for an antigen or antibody. The purpose is to provide a kit for use.

[0006]

The present invention is characterized in that a sintered molded member is used to detect an agglutination reaction to detect an antigen or an antibody in a measurement sample.

In the present invention, a detector equipped with a sintered molded member is used to separate the agglomerated reaction product from the non-agglomerated reaction product, but the value of the nominal filtration accuracy of the member allows unaggregated particles to pass through. Any size may be used. Further, members having different nominal filtration accuracy may be combined. Here, the nominal filtration accuracy is the minimum particle size that can be captured when actually filtering using a member. Further, the member can be impregnated with a solvent in order to facilitate the movement of the measurement sample. As the solvent, any known solvent for agglutination test can be used. For example, water, physiological saline, various buffer solutions (phosphate buffer solution, Tris buffer solution, glycine buffer solution), albumin solution, dextran solution, normal human and animal serum solution, synthetic polymer solution, surfactant aqueous solution And combinations thereof. Further, the sintered molded member may contain an antibody (or an antigen) corresponding to the antigen or the antibody in the measurement sample, respectively.

The material of the member used for filtration is not particularly limited as long as it can be sintered and molded. Normal,
Olefinic high polymers, polypropylene, soft polyethylene and hard polyethylene are used. In the present invention, the device for detecting the agglutination reaction can have various shapes. The sintered molded member may be placed in a container such as a transparent test tube or a transparent cell, or may have any other desired shape. Further, the sintered molded member itself may be covered with an exterior member such as a transparent plastic film except for the surface that directly contacts the sample. The container or the plastic film and the sintered molded member are welded or bonded by an appropriate method so that the sample does not pass through the gap. The material of the container and the exterior member is preferably the same as that of the sintered molded member.

The method of the present invention is a method for detecting an antigen or an antibody by separating an aggregate that aggregates by an antigen-antibody reaction from the sample and measuring one or more of the aggregate reaction product and the non-aggregation reaction product, (A) An antigen or an antibody is brought into contact with the antigen or an antibody corresponding to the antibody or a carrier particle having the antigen on the surface to form an agglutination reaction product, and the aggregation reaction product is formed in the step (a). A sample containing the agglutination reaction product is passed through the sintered molding member, and the agglomeration reaction product is captured in the sintering molding member, while the non-aggregation reaction product is allowed to pass therethrough, and A method for detecting an antigen or an antibody, which comprises detecting the presence or absence of one or more of an object. In the method of the present invention, it is possible to form the agglomeration reaction product in advance and then pass the sample through the sinter-molded member, but it is also possible to form the agglomeration reaction product in the sinter-molded member. In that case, an antibody or an antigen respectively corresponding to the antigen or the antibody in the measurement sample is included in the sintered molded member.

The particles used in the present invention are particles having an antibody or an antigen on the surface, and the particles are preferably red blood cells, antigen (or antibody) -sensitized red blood cells or antigen (or antibody) -sensitized latex. To be When a latex is used, it is convenient to use a colored material or a colored material because the agglutination reaction can be visualized. The particle size of the latex is not particularly limited, and those generally used in the art can be used, but 0.
1-10 μ is convenient.

In the method of the present invention, the formed agglutination reaction product can be separated from the non-aggregation reaction product by natural sedimentation, but centrifugation is preferred in order to save time. Moreover, you may utilize a capillary phenomenon.

As a preferred embodiment of the present invention, (a) a latex suspension prepared by sensitizing an originally colored or colored latex with an antibody corresponding to an antigen in a measurement sample or an antigen corresponding to an antibody in a measurement sample. A detection kit comprising a suspension and (b) a detection device provided with a sintered molding member for detecting an agglutination reaction can be provided. The kit may also include a diluent. Any known solvent for agglutination test can be used as the diluent.

To carry out the measurement using the kit, for example, the following is carried out. A sample containing an antigen and an antibody-sensitized latex suspension corresponding to the antigen in the measurement sample are mixed and reacted. The sample may be diluted with a suitable diluent prior to mixing. Conditions under which the mixed sample is put into a transparent tube provided with a sintered molding member, or mixed on a transparent tube provided with a sintered molding member in advance, and after the reaction, the agglutination reaction product and the non-aggregation reaction product can be separated. Centrifuge at. The centrifugation conditions may be selected as appropriate. After centrifugation, the presence or absence of one or more of the agglutination reaction product and the non-aggregation reaction product is visually confirmed.

In carrying out the present invention, the object to be measured is not particularly limited, and any object can be used as long as it can be measured by utilizing the particle aggregation reaction. For example, blood group test, antiglobulin test, HBs, TPHA, CMV, H
Examples include virus infection tests such as CV, HIV, HTLV, and tumor marker tests such as AFP, CEA, PSA, CA19-9.

[0015]

The sintered molded member used in the present invention is usually used to remove particulate impurities from air or liquid.
The member has a porous structure, and particles larger than the nominal filtration accuracy value are trapped on the surface of the member or while passing through the member. According to the method of the present invention, the agglutination reaction product due to the antigen-antibody reaction is trapped while passing through the sintered molded member, and the non-aggregated particles pass through. As a result, it becomes possible to objectively determine the presence or absence of aggregation and its intensity.

[0016]

EXAMPLES The present invention will now be described with reference to examples.

Example 1 As a sintered molded member, filter lens R (Kanken Co., Ltd.)
To use. 1 to 3 show an example in which a transparent tube, a rectangular parallelepiped transparent cell, and a triangular transparent cell are used as a reaction container, and the sintering molded member 12 is provided in the container. A shape other than the shapes shown here can be freely selected.

Example 2 FIG. 4 shows an example in which the sinter-molded member of Example 1 is used, a reaction section 14 is provided on the top of the sinter-molded member, and the reaction container and the sinter-molded member are integrally formed. Show.

Example 3 FIG. 5 shows an example in which the same sintered molded member as in Example 1 was used and a void 16 was provided in the lower portion of the container. The provision of the void facilitates the determination. The sintered molded member 12 and the void 16 may be filled with the aqueous solution. Further, as shown in FIG. 6, a threshold 15 may be provided inside the container, the sintered molded member 12 may be placed in one of the chambers, and the unaggregated matter may move to the other chamber 17. .

Example 4 FIG. 7 shows members having different nominal filtration accuracy (filter lens R
▼) An example in which 18, 20, 22 are combined is shown. The combination makes it easier to determine the presence or absence of aggregation and the degree of aggregation.

Example 5 FIG. 8 shows an example of detecting an agglutination reaction product by utilizing the capillary phenomenon. Sample liquid 28 containing aggregates reacted in the tube
The tip of the separation material 24 for detecting agglutination reaction is dipped in it and sucked up for determination. At this time, if the sample amount is small, the sample is not sufficiently sucked up. Therefore, after the sample is absorbed, the sample is immersed in a buffer solution or the like for further absorption. For this absorption, it is convenient to combine a polymer absorbent 32 as shown in FIG. As the polymer absorbent, a conventionally known one is used. In addition, in order to facilitate the initiation of the capillary phenomenon, FIG.
As described above, it is preferable to combine the sample introducing member (filter material having coarse mesh) 34. The material of the introducing member is not particularly limited.

[0022]

[Effect] According to the present invention, it is possible to provide an agglutination reaction detection device that does not require special consideration during storage or transportation. In addition, since it is molded, it can have any shape.
Further, if the particle diameters of the materials are made uniform, reproducibility is improved and the difference between lots can be suppressed. In addition, a separation material suitable for various particle sizes can be manufactured by changing the type of material (particle size or material). Further, the capillary phenomenon can be easily realized.

[Brief description of drawings]

FIG. 1 is a vertical sectional view of an antigen or antibody detection device provided with a sintered molded member in a transparent tube in Example 1 of the present invention.

FIG. 2 is a vertical cross-sectional view of an antigen or antibody detection device in which a sintered molded member is provided in a transparent cell of a rectangular parallelepiped in Example 1 of the present invention.

FIG. 3 is a vertical cross-sectional view of an antigen or antibody detection device provided with a sintered molded member in a triangular transparent cell in Example 1 of the present invention.

FIG. 4 is a vertical cross-sectional view of an antigen or antibody detection device in which a reaction container and a sintered molded member are integrally formed in Example 2 of the present invention.

FIG. 5 is a vertical cross-sectional view of an antigen or antibody detection device provided with a void in the lower portion of the container in Example 3 of the present invention.

FIG. 6 is a vertical cross-sectional view of an antigen or antibody detection device having a threshold inside a container in Example 3 of the present invention.

FIG. 7 is a vertical cross-sectional view of an antigen or antibody detection apparatus in which sintered molded members having different pore diameters are combined in Example 4 of the present invention.

FIG. 8 is a diagram for detecting an agglutination reaction product by utilizing a capillary phenomenon in Example 5 of the present invention.

FIG. 9 is a vertical cross-sectional view of a separation material for detecting an agglutination reaction, which is a combination of a polymer absorbent and a sintered molded member in Example 5 of the present invention.

FIG. 10 is a vertical cross-sectional view of a separation material for detecting agglutination reaction in which a coarse member and a sintered molded member are combined in Example 5 of the present invention.

[Explanation of symbols]

 10 Reaction Vessel 12, 18, 20, 22 Sintered Molded Member 14 Reaction Part 15 Threshold 16 Gap 17 Unaggregate Moving Part 24 Separation Material for Agglutination Reaction Detection 26 Exterior Member 28 Sample Liquid Containing Aggregation Reaction Product 30 Lid Material 32 Polymer absorbent 34 Introduction member

Claims (16)

[Claims] 1. An apparatus for detecting an antigen or an antibody by separating an agglutinate aggregated by an antigen-antibody reaction from a sample and measuring one or more of the agglutination reaction product and the non-aggregation reaction product, wherein the sample is contained. 1. An apparatus for detecting an antigen or an antibody, comprising: a reaction container according to claim 1, and a sintered molding member for separating aggregates in a sample contained in the reaction container. 2. The apparatus according to claim 1, wherein the reaction vessel and the sintered molded member are integrally formed. 3. The apparatus according to claim 1, wherein the sintered molded member is a combination of filter media having different nominal filtration accuracies. 4. The apparatus according to claim 1, wherein the sintered molded member is impregnated with a solvent. 5. The device according to claim 1, wherein the sintered molded member contains an antigen or an antibody. 6. The apparatus according to claim 1, wherein the sintered molded member is at least one selected from olefin high polymers, polypropylene, soft polyethylene and hard polyethylene. 7. A method for detecting an antigen or an antibody by separating an agglutinate aggregated by an antigen-antibody reaction from a sample and measuring one or more of the agglutination reaction product and the non-aggregation reaction product, wherein (a) the antigen Alternatively, the agglutination reaction product is formed by contacting the antibody with the antigen or an antibody corresponding to the antibody or a carrier particle having an antigen on the surface to form an agglutination reaction product, and (b) the agglutination reaction product formed in step (a). The sample containing is passed through the sintered molded member to capture the agglomerated reaction product in the sintered molded member, while allowing the unaggregated reaction product to pass therethrough, and (c) one of the agglomerated reaction product and the unaggregated reaction product or A method for detecting an antigen or an antibody, characterized by detecting the presence or absence of many. 8. A method according to claim 7, wherein step (a) is carried out in a sintered molded part. 9. The method according to claim 7, wherein step (b) is performed by centrifugation. 10. The method according to claim 7, wherein the step (b) is performed by capillarity. 11. The method according to claim 7, wherein the carrier particles are red blood cells. 12. The method according to claim 7, wherein the carrier particles are antigen-sensitized red blood cells or antibody-sensitized red blood cells. 13. The method according to claim 7, wherein the carrier particles are an antigen-sensitized latex or an antibody-sensitized latex. 14. The method according to claim 13, wherein the latex is originally colored or colored. 15. A test kit for detecting an antigen or an antibody in a measurement sample using a particle agglutination reaction,
The kit comprises (a) a latex suspension obtained by sensitizing an originally colored or colored latex with an antibody corresponding to an antigen in a measurement sample or an antigen corresponding to an antibody in a measurement sample; A detection kit comprising a detection device equipped with a sintered molded member for detecting an agglutination reaction. 16. The detection kit according to claim 15, which contains a diluent.