JP3290730B2 - Apparatus and method for developing a reaction configuration - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to an apparatus for analyzing the presence of a specific substance based on the dispersibility of reactive fine particles. More specifically, the present invention relates to an apparatus for analyzing a specific substance in a test sample in a clinical test or research. The present invention relates to an apparatus and a method for developing a reaction form for developing aggregates and non-agglomerated particles formed in the presence of a different development form that can be distinguished from each other.

[0002]

2. Description of the Related Art A technique for judging the presence of a predetermined test substance based on the presence or absence of agglutination based on an agglutination reaction has been known for a long time. A microtiter method for optically measuring the morphological characteristics of the deposited fine particles may be used. In the microtiter method, usually, a reagent solution containing a predetermined amount of fine particles having a particle size of 1 to 20 μm and a test sample are dispensed into a concave well having a hemispherical or conical bottom, and the fine particles are placed on the bottom. The presence of a specific antigen or antibody in a test sample is detected by morphologically discriminating the state of deposition on the sample. Since the reaction mode developed on the bottom of the concave well shows concentric expansion, the presence, size,
The reaction result can be determined based on morphological features such as clarity and lightness. Japanese Patent Publication No. 2-16875 describes an automatic analyzer for performing a microtiter method using a microplate having a concave well.

[0003] As a conventional improvement technique relating to the microtiter method, there is a technique in which a concave well is centrifuged or fine particles having a large specific gravity are sedimented under natural gravity, thereby accelerating the sedimentation speed and shortening the analysis time. . When making fine particles heavy or creating high gravity, the force in the direction of gravity acting on the fine particles is averagely amplified at any position, so that all the fine particles sink in the reaction solution regardless of aggregation or non-aggregation. As well as rolling easily on the inclined bottom surface. Therefore, in order to develop different reaction modes depending on the presence or absence of aggregation, various conditions such as the shape or inclination angle of the inclined bottom surface, the specific gravity or particle size of the fine particles, and the centrifugal speed are restricted, and the analysis time is reduced. It cannot be expected to be significantly reduced.

In Japanese Patent Application Laid-Open No. Hei 2-124464, an agglutination reaction is caused by bringing a magnet having a flat magnetic pole surface oriented in a horizontal direction close to a lower part of a concave well having a hemispherical or conical inclined bottom surface. A method of magnetically attracting magnetically responsive fine particles toward an inclined bottom surface is disclosed. Here, the magnetic pole surface oriented in the horizontal direction emits a magnetic flux along the direction of gravity substantially toward the bottom of the well, while the magnetic attractive force is directed from the high position to the low position of the inclined bottom surface. A magnetic force gradient is formed such that the magnetic force gradually increases. Therefore, in such a magnetic field environment, the aggregated particles attracted to the entire surface on the inclined bottom surface are appropriately stopped, and the tendency of the non-aggregated particles that are easily rolled to efficiently gather at the lowest part is brought about. Each reaction mode developed with the fine particles can be clearly different. The agglutination reaction using magnetic attraction surprisingly surpassed the conventional measurement sensitivity. Moreover,
The time until the reaction form is completed by magnetic attraction is
Compared to the technology using centrifuges and high specific gravity particles, it has achieved a speed of less than one-fourth. Furthermore, by utilizing magnetic attraction, even when using a blood sample containing red blood cells,
A reaction form based on agglutination can be developed before red blood cells sediment.

On the other hand, there is also known a technique in which an agglomeration phenomenon between fine particles by an agglutination reaction is performed on a flat slide glass. Are determined as non-aggregation if the fine particles are monodispersed uniformly over the entire plane, and aggregation if the aggregates are sparsely scattered. JP-A-4-2088836 discloses a reaction vessel in which two flat transparent plates are inclined at a predetermined angle while facing each other at an interval such that a capillary phenomenon occurs, and the reaction vessel contains fine particles subjected to an agglutination reaction. A small amount of the reaction solution is held between the flat plates, and only the non-aggregated fine particles are rolled on the inclined surface, thereby developing a different band-like reaction mode between the aggregation and the non-aggregation. At this time, the capillary phenomenon dramatically increases the contact efficiency of the reaction solution with the inclined surface,
It realizes highly sensitive analysis even with a small amount of reaction solution.

The present inventors have already disclosed in Japanese Patent Laid-Open No. 4-36947.
No. 7 discloses a technique in which a reaction form of magnetically responsive fine particles accompanying an agglutination reaction is developed on a flat wall surface of a reaction support along the horizontal direction. That is, a magnet having a hemispherical or conical convex surface as a magnetic pole is positioned below the flat surface. Specific configurations of the convex magnetic pole surface include a case where the magnetic pole surface of one permanent magnet is molded and a case where a plurality of magnetic pole surfaces of the electric magnet are arranged in a step shape. In addition, as a method of not performing such molding, a flat magnetic pole surface is inclined at a predetermined angle with respect to a reaction support having a flat surface, thereby developing a strip-shaped reaction mode that can be determined. On a horizontal flat surface, it is possible to develop a reaction mode depending only on the magnetic attraction, so that it is possible to relatively easily obtain a result based on the agglutination reaction.

[0007]

In the technique of depositing predetermined fine particles on the concentric inclined bottom surface, the aggregated fine particles are liable to collapse to the lower inclined portion due to their own weight with time, and are erroneously determined to be non-aggregated. Sometimes. Further, when the shape of the inclined bottom surface varies depending on the manufacturer, product number, lot, etc., the rolling characteristics of the fine particles fluctuate, so that it is difficult to obtain reactivity under a certain condition, and the determination accuracy is reduced. . In addition, in the prior art which promotes the moving speed of the fine particles, if the inclination angle of the bottom surface of the reaction support varies, a stable reaction form cannot be obtained. Further, in the conventional method of performing an agglutination reaction on a flat surface, it is difficult to develop an easily identifiable reaction form between agglomerated particles and non-agglomerated particles. Further, in the prior art, since all particles are deposited in the direction of gravity, erythrocytes or other contaminants that can be deposited obscure the reaction form, which makes it extremely difficult to identify the reaction form. Some may show false positives and / or negatives. Moreover, although the strip-shaped reaction mode developed on the horizontal plane is relatively stable against gravity, it is unknown how to obtain a reaction mode that is easy to determine.

An object of the present invention is to provide an apparatus and a method for developing a reaction mode which has a stable reactivity and a high determination accuracy even for reaction supports of various different shapes. Another object of the present invention is to provide an apparatus and a method for developing a reaction mode that can always be accurately and easily determined without being affected by various sedimentary contaminants.

[0009]

SUMMARY OF THE INVENTION In view of the above-mentioned problems, the present invention has a reaction support having a plane and / or curved surface which is inappropriate for discriminating the presence or absence of an agglutination reaction. Therefore, the present invention provides an apparatus and a method for solving the above-mentioned object by providing a point-symmetrical, and preferably concentric, reaction form even when the reaction is performed.

[0010] That is, the present invention develops a required amount of fine particles.
Curved or flat surface that has an area large enough to fit and contains a straight line
A reaction support having a development region, and a magnetic generator whose magnetic flux direction is inclined with respect to a vertical axis of the development region , wherein at least two of the magnetic generators are used for the reaction. The same deployment area, including the straight part of the support
A magnetic force gradient symmetrical with respect to the region is provided . Distance magnetic generator and a straight line on the reaction support, when configuring gradually away orientation relationship radially from the center of the point symmetry, the virtual plane perpendicular to the shape or flux of the pole faces, It has a roof shape or a polygonal pyramid or cone shape. In addition, the present invention
A song that has an area enough to spread the fine particles and includes a straight line
For reactions that have a flat or flat deployment area
A support, and a magnetic generator;
Acts on the same deployment area, including the line sections
And the two magnetic generators are connected to each other so that the magnetic flux is connected.
Point symmetry with respect to the deployment area with different magnetic poles
It is characterized by being arranged so that a strong magnetic gradient occurs.
It is. At this time, the magnetic generators of different diameters
It is good to arrange | position between the expansion | deployment area | regions of the support body for use. Further
Has an area enough to spread the required amount of fine particles, and
A development area that is a curved or flat surface containing a straight line
A reaction support having a magnetic generator
A magnetic part in which the generator is positioned near the deployment area
Material and a magnet for magnetizing the magnetic member,
These magnetic member and magnet are used as the support for the reaction.
A point-symmetric magnetic gradient is created for the same deployment area.
It is characterized by being arranged as follows.

Further, the present invention provides a method for developing a required amount of fine particles.
Curved or flat surface that has an area large enough to fit and contains a straight line
A reaction support having expansion area as is, a magnetic generator which is tilted magnetic flux direction relative to the vertical axis of the expansion area, the linear portion of the reaction support the magnetic generator
Point-symmetric rotation about the same unfolding area including the minute
Moving means for moving in an inclined state along the trajectory .

Further, the present invention provides a step of reacting a group of magnetically responsive fine particles having a fixed amount of a substance that specifically binds to a test substance with a sample in which the presence of the test substance is unknown. A step of magnetically moving the group of microparticles after the reaction toward a predetermined wall surface, and a step of expanding the group of microparticles moved to the wall surface in an area for expansion along the predetermined wall surface; and Test substance based on morphology
And a step of identifying the presence, absence or degree of reaction with,
The expanding step includes a step of forming a point-symmetric magnetic force gradient with respect to the same developing area of the wall surface, and unreacted fine particles that have reached the wall surface are expanded toward the point of symmetry of the magnetic force gradient. It is characterized by the following.

The test samples treated in the present invention mainly include blood,
Biological materials suitable for obtaining medical information in animals (especially mammals including humans), such as urine and tissue pieces, are included. When the surface antigen of particle-like cells having dimensions, colors, and shapes that can be easily detected optically, such as red blood cells, is to be tested, a magnetic substance in an amount sufficient to provide magnetic responsiveness to the particle-like cells And a reagent linking the antibody to the surface antigen. Optical detection, such as hormones, enzymes, viruses, bacteria, allergens, and various antibodies, when the substance itself is smaller than fine particles and the presence or absence of aggregation is a size that is difficult to detect with the naked eye or optical measuring instruments Aggregates of magnetic microparticles are formed by adding a reagent that has a possible particle size and is a liquid containing a predetermined amount of magnetically responsive microparticles (hereinafter referred to as magnetic microparticles) bound to a predetermined antigen or antibody. Is done. For cell samples that have dimensions that allow easy optical detection, such as platelets, various lymphocytes, tumors, and cultures, but have poor particle and light absorption properties, fix them on a predetermined reaction support. A cell layer may be formed, and magnetic fine particles may be reacted on the cell layer.

The agglutination reaction to which the present invention is applied includes a direct agglutination method in which the test object itself is agglutinable particles and a specific particle (for example, artificially synthesized particles, treated blood cells) Etc.) are largely divided into the indirect agglomeration method of forming agglomerates. As a reaction system, a known amount of a competitor capable of agglutinating with the magnetic fine particles in the reagent as well as the test substance is reacted with the reagent together with the unknown test sample, A sandwich system using a reaction support on which a specific reactive antigen or antibody is immobilized may be used. In particular, the sandwich system is also called a solid phase agglomeration method because it includes a step of binding an object to be tested in a sample to a support to form a solid state, and for example, an indirect Coombs method, a mixed passive agglutination method (MPHA) ) Etc. correspond to this.

The shape of the reaction support is such that the area where the aggregates of the aggregated magnetic fine particles are distributed by magnetic attraction is substantially larger than the area occupied by the group of magnetically aggregated non-aggregated magnetic fine particles. Any shape, size and material can be applied as long as it is a non-magnetic material having an area and a magnetic permeability. However, in order to more efficiently identify the reaction mode formed on the reaction support, it is preferable to apply a member that imparts light transmittance or light reflectivity to the reaction support. However, variously selecting or designing a reaction support so as to be advantageous in terms of economy, easiness of measurement, and the like, appropriately implements the configuration and operation and effect of the present invention within an obvious range. It does not depart or limit the technical scope of the invention.

For example, when using a reaction vessel having a concave space capable of storing a fixed volume of liquid, such as a known test tube, cuvette, microtiter plate, etc. The magnetic particles can be distributed by positioning a predetermined magnet at the position.
When a hollow body having an inlet opening and an outlet opening, such as a flow cell or a capillary vessel, is used as a reaction support, a predetermined reaction solution is introduced into the hollow body through the inlet opening and a predetermined amount of magnetic material is introduced. A similar distribution can be formed by positioning the magnet outside the hollow body in the presence of the fine particles. When using a plate-like reaction support consisting of a flat glass or a curved surface or a flat surface having no side surface capable of holding a liquid, the plate-like reaction support is placed in a liquid containing a predetermined amount of magnetic fine particles. By immersing and incubating under a magnetic force distribution as described above for a predetermined time, a distribution of magnetic fine particles can be formed on at least one surface of the plate surface. When the wall surface of the reaction support has a porous filter or a large number of fine projections, the movement of non-agglomerated particles and / or aggregates is selectively restricted during magnetic movement, and This is preferable because the morphological difference between aggregation and aggregation can be increased.

The magnetic fine particles used in the present invention can be arbitrarily selected from various known and commercially available materials and shapes, but preferably contain a magnetic material which does not substantially generate remanence. Here, the amount of the magnetic substance contained in the magnetic fine particles as a reagent can be adjusted to such an extent that the aggregate to be analyzed is magnetically moved in the reaction solution, so that a reaction form based on aggregation can be created. It is preferable to set the optimum amount as needed in consideration of the balance of the moving time, the distribution forming time, and the like. Only when the magnetic fine particles are moved along the direction of gravity, the specific gravity of each magnetic fine particle or agglomerate may have a sedimentation speed exceeding the speed of movement by the magnet. If the behavior of the microparticles is under magnetic conditions that are determined solely by magnetic force rather than gravity, the desired form of reaction is obtained.

As a typical example, when the method and apparatus of the present invention are carried out using a commercially available microtiter plate, the configuration of the magnetic fine particles may be spherical or polygonal having an average particle diameter of 0.1 to 50 μm. And a polymer or a coacervate containing a predetermined amount of ultrafine particles having a particle size of 0.1 μm or less. However, the average particle diameter of the magnetic fine particles is 3 to 15 μm in consideration of the contrast of the reaction form due to aggregation and non-aggregation.
It is particularly preferable to select from the range of 5 to 12 μm. The specific gravity of the magnetic fine particles is preferably in the range of 1.1 to 3.0, particularly preferably 1.1 to 1.5. When using magnetic particles having a low specific gravity or a buoyancy such as ultrafine particles or hollow particles having a particle size of 1 μm or less, a relatively strong magnetic force (preferably 3,000
Gs or more) to avoid the influence of buoyancy. On the other hand, in the case of magnetic fine particles having a high specific gravity exceeding 1.5, a relatively weak magnetic force (preferably 500 to 1,000
0 Gs or less). On the other hand, when the magnetic fine particles are magnetically moved in a direction other than the direction of gravity, a strong magnetic force (for example, 1,500 Gs or more) is applied at least so as not to be affected by the weight of the fine particles.

The amount of the magnetic substance contained in the magnetic fine particles is usually such that a light-transmissive reaction support provides a dark brown color which is convenient for detection by the naked eye or an optical measuring instrument after the formation of an aggregated or non-aggregated particle distribution. I do. However, if the magnetic fine particles in the reagent used for the reaction with the test sample have a small particle size and / or low particle concentration for micro-measurement or high-sensitivity analysis, the reaction and distribution formation may be hindered if necessary. The content of the magnetic substance in the reagent may be increased to such an extent as not to exceed, or magnetic fine particles containing an appropriate commercially available dye or a dark material (such as carbon powder) as an additive may be prepared. In addition, in order to improve detection sensitivity, an appropriate tracer of a fluorescent substance, a luminescent substrate, a chromogenic substrate, an enzyme, a radioisotope, etc. is bound to the magnetic fine particles at the latest before detection of the agglutination reaction. Thus, the S / N ratio at the time of detection may be improved. A specific example of such a tracer may be any commercially available one. However, when a colored substance such as red blood cells is contained as a reaction component, it is preferable to encapsulate a minimum necessary magnetic substance in the fine particles.

The magnetic generator according to the present invention means a permanent magnet or an electric magnet having a magnetism that attracts magnetic fine particles for reaction at least to the extent of overcoming gravity. When a permanent magnet is used, it may be appropriately selected from commercially available or known magnets having a predetermined coercive force, such as ferrite and rare earth cobalt, in consideration of economy, workability, magnetic properties, and the like. . The magnetic force required for the magnet can be the same as the effect provided that the magnetic attraction reaches all the magnetic fine particles provided to the reaction support, so that the desired reaction time and moving time can be obtained. May be changed as appropriate. In order to move magnetic fine particles having an average particle size of 0.1 to 50 μm in a liquid, a magnetic force in an effective range of about 50 to 10,000 gauss only needs to act on the fine particles, but the time is reduced while securing the required reactivity. 200 to 5,000 gauss, especially 500 to 3,000 gauss, is suitable. The angle of inclination when the magnetic pole surface to be employed is inclined may be determined by appropriate experiment according to various conditions such as the viscosity of the liquid containing the magnetic fine particles, the surface state of the reaction support, and the morphological characteristics before and after the aggregation of the magnetic fine particles. Change the settings. Further, the separation distance between the magnet and the reaction support may be arbitrarily selected so that the reaction time becomes appropriate as long as the prepared magnetic force gradient sufficiently acts.

When an electric magnet consisting of a conductive coil and an appropriate electromagnetic circuit is used in place of the permanent magnet, the magnetic gradient can be changed by electric control without changing the shape of the magnet. This is preferable because it can effectively cope with item inspection. Further, it is suitable for automation in that application of magnetic attraction can be reduced only by turning on / off the power without moving the magnet. If necessary, a configuration for adjusting the temperature of the surface of the electric magnet by an appropriate cooling means or a heat insulating member may be added so that the Joule heat generated from the electric magnet does not change the reactivity of the reaction solution. .

The magnetic pole of the magnet may be either the N pole or the S pole.
No. as disclosed in the above item. However, when multiple magnets are arranged in the same time with respect to the reaction support, the arrangement relationship of each magnet and the application timing should be changed in design in consideration of the case where the magnetic flux of the magnets may interfere with each other. It is preferable to secure a desired magnetic force gradient by, for example, sufficiently separating a plurality of reaction supports. In addition, when a plurality of magnetic pole surfaces are combined, various magnetic flux lines are distributed depending on whether the magnetic poles are the same magnetic pole or different magnetic poles. Therefore, the magnetic flux lines inclined in a point-symmetric arrangement on a plane or a curved surface of the reaction support. If the distribution occurs, the operation and effect of the present invention can be obtained. In addition, the magnetic force gradient accompanying the inclination of the magnetic flux lines forms a magnetic attractive force that converges toward the center if it is convex with respect to the reaction support, and diffuses radially from the center to the periphery if it is concave. Since such magnetic attraction can be formed to form a point-symmetric reaction mode in any case, a convex or concave shape may be selected as appropriate in consideration of the type of the reading means and the ease of identification. Furthermore, if the inclination angle formed by each of the plurality of magnetic pole faces is experimentally selected so as to be an effective angle range appropriately according to the measurement item, the property of the test sample, the rolling characteristics of the fine particles, etc., the optimum reaction is always obtained. It can be set to complete a reaction mode that can be determined after a lapse of time. For example, in order to make it easier to clearly identify the reaction mode of the magnetic fine particles accompanying the agglutination reaction by the presence or absence of the reaction, the direction of the magnetic flux should be at least with respect to the wall surface of the reaction support that is to form the distribution of the agglomerates. What is necessary is just to apply a symmetric magnetic force gradient to one point.

As one of such methods, the magnetic fine particles after the reaction are provided in at least two directions by applying a magnetic flux density on the wall surface of the reaction support so as to increase or decrease in at least two directions. There is a method of forming a magnetic attraction such that the magnetic attraction is developed symmetrically. Such magnetic attraction is achieved by inclining the magnetic flux direction with respect to the reaction support. Specifically, FIGS. 1A and 1B
As shown in the figure, the magnetic pole surfaces of two magnets 1 and 1 ′ having the same area and emitting the same density of magnetic flux vertically are separated at the same angle so that they are adjacent to each other as one composite magnet in line symmetry. The attached composite magnet 2 is exemplified.
Since the two magnetic pole surfaces that have been joined emit magnetic fluxes inclined in different directions in a line-symmetric manner, an attractive force that gradually converges in a line-symmetric manner is applied to the magnetic fine particles on the reaction support. At this time, the aggregated magnetic fine particles hardly roll on the reaction support, while the non-aggregated magnetic fine particles roll almost without hindrance, and converge toward the linear converging region.
Therefore, if the state of distribution of the magnetic fine particles that have undergone the agglutination reaction develops on the support for reaction is morphologically distinguished, it can be easily identified whether or not they have aggregated.

In FIGS. 1A and 1B, even if the composite magnet is divided into a plurality of magnets and then joined, each magnet has an N pole and an S pole, and therefore, one magnet Can produce a plurality of composite magnets. Also,
The same tendency is basically exhibited regardless of whether the magnetic pole surface to be bonded is a combination of an S pole and an N pole or a configuration having only the same magnetic pole. However, considering the magnetic interference between the magnetic poles of the composite magnet, it is better to arrange different magnetic poles adjacent to the joining part, because the magnetic flux between the adjacent magnetic poles is connected to form a high-density magnetic field This is preferable in that the reaction force can be more clearly identified since the reaction form can be more clearly identified.

Similarly, a composite magnet in which three or more magnetic pole faces are arranged point-symmetrically has a polygonal pyramid shape on the magnetic pole face. Bring to the support. Therefore, when a composite magnet having three or more magnetic pole faces is opposed to a flat or curved surface of the reaction support, three or more magnetic pole faces are required depending on the number of magnetic pole faces to be joined.
A reaction mode of magnetic fine particles radially symmetric with respect to the above directions is formed. It is preferable that the reaction form be radially symmetrical in more directions, since it becomes obvious to the observer.

When a magnetic generator having a line-symmetric magnetic gradient is opposed to a reaction support having a line-symmetric wall surface, a concentric magnetic focusing action acts on the magnetic fine particles. For example, FIG. 2A shows that the composite magnet 2 shown in FIG.
3b is a state where the rectangular cuvette 4 is disposed so as to be orthogonal to the right-angled side surface. In such an arrangement, the magnetic fine particles contained in the cuvette 4 are concentrically converged and moved by the synergistic action of the right-angled cuvette wall surfaces 5a and 5b and the right-angled magnetic pole surfaces 3a and 3b. Therefore, the non-aggregated magnetic fine particles converge at the intersection of the orthogonal side surfaces as shown in FIG. 3A and develop so as to form a button-shaped reaction form, while the aggregated magnetic fine particles are formed as shown in FIG. 3B. Develop a reaction pattern that spreads out in a circle. The concentric reaction form facilitates both the naked eye and the judgment by the judgment device. FIG. 2 (B) and FIG.
FIG. 2C is an example in which the configuration of FIG. 2A is modified, in which a columnar magnet 7 formed of one hemispherically shaped magnetic pole surface is orthogonal to a hemispherical side surface of a cylindrical test tube 6. FIG. 2 shows a case where the composite magnets 8 are joined so that the magnetic pole surfaces 9a and 9b are adjacent to each other at an obtuse angle, and FIG.
The same operation and effect as in (A) are shown.

As shown in FIG. 2, when utilizing the synergistic effect of the side surface shape of the concave container as the reaction support and the magnetic force gradient of the composite magnet as the magnetic generator, the side surface of the concave container is formed. The angle can be changed variously. That is, if the side surface angle of the concave container is less than 180 degrees at an obtuse angle of less than 90 degrees or greater than the spherical surface, if the angle formed by the pole faces of the composite magnet is also obtuse or the curvature is increased, the Since the convergence force of the magnetic particles on the magnetic fine particles is reduced, the method is suitable for detecting a test substance that forms an aggregate having a weak binding force or a relatively small aggregate. Conversely, if the side angle of the concave container is more acute than 90 degrees or the curvature is smaller than the spherical surface, if the angle formed by the magnetic pole surfaces of the composite magnet is made more acute or the curvature is smaller, the reaction solution contains Since the convergence to the magnetic fine particles is emphasized, the method is suitable for speeding up the detection of a test substance that forms a relatively large aggregate, particularly an aggregate that is difficult to roll on the container side wall. In any case, as shown in FIG. 2 (A), if both the reaction support and the magnetic generator are a combination of angular shapes, a conical focusing force is generated synergistically. It is possible to develop a reaction mode substantially equivalent to that of the concave well having the bottom surface. Further, as shown in FIG. 2B, if both the reaction support and the magnetic generator are a combination of curved surfaces,
Since a spherical converging force of at least a part of a true sphere or an elliptical sphere is generated synergistically, a reaction mode substantially equivalent to that of a concave well having a conventional hemispherical bottom surface can be developed.

A point-symmetric magnetic gradient can also be formed by rotating one inclined magnet about the point of symmetry. Specifically, FIG. 4 (A),
As shown in FIG. 1B, a rotating plate 11 located immediately below a concave reaction vessel 10 having a flat bottom surface is connected to a motor 12 controlled to a desired rotating behavior by drive control means (not shown). For example, a device in which an arc-shaped magnet 14 having a magnetic pole surface 13 inclined with respect to the rotating portion is fixed to an arc portion of the rotating plate 11. Here, the upper surface 13 of the arc-shaped magnet 11 is a magnetic pole of either N or S, and the magnetic flux generated from this magnetic pole extends substantially parallel while maintaining a constant inclination at least until passing through the bottom surface of the reaction vessel 10. Shall be Further, the rotating plate 11 is rotated at a desired speed by the motor 12 and drive control means (not shown), for example, at an appropriate number of rotations of 10 to 200 rotations per minute. According to such a rotary moving unit, the strongest magnetic force acts on the central portion of the bottom surface of the reaction vessel 10 near the center of the rotary plate 11 corresponding to the uppermost portion of the arc-shaped magnet 11, and concentric circles with the rotating operation. A magnetic force gradient that gradually decreases toward the outer periphery acts on the bottom surface of the reaction vessel 10. Therefore, the group of magnetic fine particles in the reaction liquid mixed or stirred in the reaction vessel 11 in the stationary state is immediately drawn to the bottom of the vessel after the agglutination reaction, and the non-magnetic particles among the fine particles that reach the bottom of the vessel are non-magnetic. The coagulated material further rolls along the magnetic gradient on the bottom of the container and gathers in the center of the container to exhibit a button-like dense reaction form, whereas the coagulated material no longer moves on the bottom of the container. It exhibits a circular reaction pattern that spreads almost uniformly without moving. Such a difference in the reaction mode can be sufficiently discriminated by the naked eye. The rotation speed may be minimized by arranging a plurality of arc-shaped magnets 14 at the same interval on the circumference of the rotating plate 11. In addition, the angle of the arc of the arc-shaped magnet 14 can be set to an acute angle as the number of magnets arranged on the rotating plate 11 increases, or as the rotation speed of the rotating plate 11 increases,
This case is preferable in that the rotation trajectory can be more approximated to a conical shape.

As a modification, a plurality of magnetic pole surfaces of the same material may be arranged in steps along the inclined surface. For example,
As shown in FIGS. 5A and 5B, a plurality of circular or polygonal ring-shaped magnets 15 and 16 having different diameters are placed on conical or polygonal pyramid-shaped supports 18 and 19 made of nonmagnetic material. It is also possible to use a magnetic generator in which concentric or radial magnetic force gradients are formed such that the magnetic force gradually increases by gradually changing the height and arranging them stepwise. Also support
A central magnet 17 having a minimum required circular upper surface as a magnetic pole is mounted on the tops of the 18, 19 so that non-agglomerated particles are easily collected. Such a step-shaped magnetic generator is mainly effective when approaching a flat reaction support, and can develop a point-symmetric reaction mode by magnetic fine particles on a flat surface. Note that each ring-shaped magnet 15, 16
Whether the upper surface of the center magnet 17 is made of different magnetic poles or the same magnetic pole may be appropriately selected according to the distance between the magnets and the magnetic flux density.

As another modified example, as shown in FIG. 6A, a pair of magnetic pole surfaces 21a of N-pole and S-pole having different areas are sandwiched by a reaction support 20 composed of a flat surface. , 21b facing each other. Here, in the space between the magnetic pole faces, a distribution of magnetic flux lines inclined in a point symmetry as shown by a dotted line is formed. The magnetic flux density provided by such a magnetic flux line distribution is low on the side where the magnetic pole surface is relatively large, and becomes asymptotically high as it goes toward the relatively small magnetic pole surface. Regarding the magnetic poles to be opposed, either side may be an N (or S) pole, or one set of magnetic poles may be constituted by magnetic poles of one magnet. The inclination angle of the magnetic flux can be easily determined by the ratio of the facing magnetic pole faces and / or the separation distance.
In addition, when a pair of magnetic pole faces are circular in shape and have different diameters and face each other in parallel so that the centers coincide, a point-symmetric magnetic flux converging in a conical shape is formed. In general, since the center of gravity of the magnetic balance is at the center of the circular magnetic pole surface, in the conical magnetic flux distribution, the magnetic fine particles move substantially along the conical magnetic flux and converge toward the center of the conical cross section. Go to If the reaction support holding the reaction solution containing the magnetic fine particles is positioned closer to the magnetic pole surface having a relatively small opposing area among the pair of opposing magnetic pole surfaces, it converges to one point of the reaction support. Magnetic attraction. At this time, since the area of the small magnetic pole also affects the aggregation area of the non-aggregated particles, the design may be changed so that the non-aggregated particles are aggregated as small as possible by appropriately tapering the tip. In order to process a plurality of reaction supports at the same time, the large side of the magnetic pole surface is constituted by a single large plate-like magnet covering all the supports, and the magnetic pole surface is sandwiched between the supports. May be opposed to each other for each corresponding reaction region. The plate-like magnet is preferably practically excellent in workability, elasticity, and thinness, such as a rubber magnet or a plastic magnet obtained by mixing rubber or plastic with magnet powder.

As shown in FIG. 6 (B), when the side having the larger magnetic pole surface is the ring-shaped magnet 22 and the small magnet 23 having the smaller magnetic pole surface is located near the reaction support 20, Since a hemispherical repulsive force is generated in the ring inner space 24, as shown by a dotted line, a conical magnetic flux forms a magnetic flux that is connected in a substantially hemispherical shape. Since the magnetic fine particles in the substantially hemispherical magnetic flux move substantially along the outer periphery of the magnetic flux and finally converge to the center of the circular magnetic pole surface, more magnetic fine particles are supported for reaction. After rolling around the body 20, it gathers at the center of the aforementioned magnetic balance. This means that more aggregated particles are more likely to stay around the support for reaction, which is preferable in that it contributes to improvement of the inspection sensitivity. In addition, if an optical system of an optical reading means is appropriately arranged on the inner space 24 of the ring-shaped magnet 22, the reaction mode can be read immediately during or after the expansion of the reaction mode of the magnetic fine particles. Also, it is preferable in that it can read a reaction support that is oriented in a direction other than the direction of gravity without being affected by gravity and stably maintaining the reaction mode of the magnetic fine particles. Conversely, if it is positioned closer to the pole face side of a relatively large area of a set of pole faces, along with a magnetic attraction such that a predetermined wall surface of the reaction support is radially diffused from one point,
Eventually, a magnetic attraction is brought about such that the magnetic flux converges at the center of the vertical section.

The direction of the magnetic attraction of a pair of opposing magnetic pole surfaces changes depending on which of the large and small magnetic poles the reaction support is closer to at an intermediate position where the magnetic attraction to the magnetic fine particles is equal. Therefore, when the reaction support is in the form of a single plate, a magnet having a large magnetic pole surface is disposed above the support and a small magnet is disposed below the support, and the support is relatively positioned. The magnetic fine particles may be attracted to the upper surface of the support by being positioned closer to the lower magnet. Here, the closer the reaction support is to the magnetic pole, the stronger the magnetic attraction to the support becomes.Therefore, by appropriately selecting the position of the reaction support positioning means, the test substance can be obtained. The desired detection sensitivity can be set at any time according to the characteristics, reaction conditions, and the like. When a pair of opposing magnetic pole surfaces are formed by an electric magnet, it is easy to increase or decrease the magnetic force of both magnetic poles at a constant ratio, so that the reaction of various volumes can be performed while maintaining the gradient of the magnetic flux. This is advantageous in that it can handle liquids.

On the other hand, of the opposing magnetic pole faces, a relatively large magnetic pole face side is made of a ferromagnetic material having little residual magnetism (for example,
Also in the case of using stainless steel, iron, etc., a conical magnetic flux distribution is generated by magnetism generated from a magnetic pole surface magnetizing a magnetic material to form a connection of magnetic flux lines. In this case, unlike a case where a pair of magnetic pole faces are opposed to each other, the magnetic attractive force acts to move exclusively in the direction of the magnetic pole face. The magnetic pole on the magnetic pole surface facing the magnetic body may be either N or S.

As another method, a magnetic pole surface of a magnet that generates a substantially uniform magnetic force in the magnetic flux direction is formed on the wall surface of the reaction support by molding the magnetic pole surface from the magnetic pole surface to the wall surface of the reaction support. It is molded so that the distance is symmetrical to at least one point, and this is opposed to a predetermined wall surface of the reaction support. The magnet may be formed by filling a powdery ferromagnetic material into a mold or shaping a permanent magnet manufactured in a predetermined shape at an appropriate angle or curvature. The angle or curvature of the magnetic pole surface to be molded may be kept constant or may be changed intermittently or continuously according to the wall shape of the reaction support to be used. However, since the magnetic flux emitted from the molded magnetic pole surface does not substantially incline with respect to the reaction support, the magnetic attraction in the oblique direction on the support is exclusively performed by the molded magnetic pole surface and the reaction support. Must be formed only by the distance relationship of Therefore,
In order to obtain the magnetic force gradient required for the present invention only by molding the magnetic pole surface, it is necessary to mold the magnetic pole surface with a relatively acute molding angle or curvature while taking into account the variation in magnetic force on the magnetic pole surface. .

Further, a plurality of parts made of different materials are used.
In the case of using a permanent magnet or a plurality of electric magnets magnetized under different voltage conditions, the point-symmetric magnetic force distribution as described above can be applied to the reaction support even if the arrangement of the individual pole faces is relatively arbitrarily configured. Can be acted upon. For example, a plurality of ring-shaped permanent magnets having different diameters and made of different materials so that the diameter and the magnetic force are correlated are formed by flattening an area corresponding to a predetermined plane or curved surface of the reaction support. Place concentrically on the surface. As described above, the use of the composite magnet in which the point-symmetric magnetic force distribution is formed while being arranged on a flat surface makes it easy to position the reaction magnet on the reaction support and also makes it possible to reduce the size of the entire apparatus. preferable. In particular, if the flat magnetic pole surface of such a composite magnet is appropriately provided with a light-reflecting color such as white or silver or a flat mirror is fitted, and the reaction support is formed of a transparent member, the action of the magnet can be achieved. Since it becomes possible to observe the reaction form of the magnetic fine particles below as a sequentially reflected image,
It is preferable because special observing means such as a photoelectric element and a CCD need not be arranged, and the space can be clearly detected in a small space in real time.

As still another method, two magnets having a line-symmetric magnetic force gradient are arranged at right angles to each other by a magnetic pole arrangement such that magnetic fluxes are connected to each other. Gradients can be synthesized. FIG. 8 (A) is described with reference to FIGS. 1 (A) and 1 (B) with a reaction support 26 in which two non-magnetic flat plates are opposed to each other with a space narrow enough to cause capillary action. A configuration is shown in which two of the composite magnets described above face each other such that the magnetic pole faces are orthogonal to each other. Here, one composite magnet 27 is adjacent to each other with N poles, and the other composite magnet 28 is adjacent to each other with S poles. In addition, each composite magnet
Reference numerals 27 and 28 have the same magnetic flux density and are arranged so as to be in contact with the approximate center of the reaction support 26 made of two parallel transparent plates, or to be spaced equidistantly in the vicinity. Further, the adjacent angle between the magnetic poles of each composite magnet is shown in FIG.
In (A), the angle is set to 90 degrees, but it may be appropriately selected from a range of more than 80 degrees and less than 180 degrees. When the rectangular composite magnets are made to cross at right angles, a magnetic force gradient substantially equivalent to the gravity acting on the conical bottom surface acts on the wall surface of the reaction support.

Here, the individual composite magnets 27,
The magnetic force gradient acting on the reaction support 26 is shown in FIG.
As shown in (1), the magnetic field has magnetic attraction that converges line-symmetrically in directions orthogonal to each other. Therefore, if these perpendicular magnetic attractive forces act simultaneously on the reaction support 26,
As a magnetic attraction is exerted on the magnetic fine particles in the reaction solution held in the support so as to concentrically converge, the magnetic fine particles in FIG.
Reaction forms as shown in (B) and (C) are obtained. In FIG. 8A, the composite magnet 27 having N poles and S
By facing the composite magnets 28 of the poles at right angles to each other, there is substantially no attractive force or repulsion between the composite magnets. This is preferable because unnecessary load is not applied and the handling becomes easy. In addition, it is preferable that each of the two composite magnets has a magnetic pole configuration as shown in FIG. Further, when the reaction is simultaneously performed on a plurality of reaction supports 26, the plurality of reaction supports are appropriately spaced and arranged in a row or in a grid pattern, and the plurality of composite magnets are vertically identical. It should just be lined up in the direction.

FIG. 9 shows an example in which upper magnetic pole surfaces 29a and 29b, which are molded so that the cross section is partially circular, and lower magnetic pole surfaces 30a and 30b are opposed to each other at right angles. . These upper and lower magnetic pole surfaces are respectively adhered to a support portion 31 supported on a rotating shaft 32 to form two rollers. Here, the rotating shaft 32 is connected to a drive source (not shown), and the lower magnetic pole surface 30a faces the upper magnetic pole surface 29a, and the lower magnetic pole surface 30b faces the upper magnetic pole surface 29b. It is assumed that the rotation is controlled in the same cycle as described above. The facing magnetic pole faces may be the same pole or different poles. The rotation speed can be arbitrarily selected from, for example, 5 to 300 rotations per minute.
In some cases, the rotation speed may be appropriately changed during the agglutination reaction, or the rotation speed may be intermittently stopped or reduced. In the case where the partially cylindrical composite magnet is made to cross at right angles, a magnetic force gradient substantially equivalent to that of gravity acting on the hemispherical bottom surface due to the magnetic synergistic effect similar to that described in FIG. Acts on the wall of the reaction support.

The distribution form of the magnetic fine particles formed on the reaction support can be identified by directly observing the reaction support with the naked eye, or by appropriately reading means known in the art, such as a CCD, a photoelectric device, a magnetic probe, or the like. It can be read as an electric signal.
The reading means may be configured to measure the amount of transmitted light passing through the reaction support or the amount of magnetization, or may be configured to measure the amount of reflected light reflected on the reaction support or the amount of magnetism emitted by magnetized fine particles. It may be. When the photometric light is transmitted through the reaction support, the reaction support is preferably made of a material having excellent light-transmitting properties.

In the case where the material of the reaction support is a non-magnetic material, as in a general reaction vessel, an appropriately designed and arranged electromagnetic generating means is used, and a single or a plurality of reaction supports are used. By applying an AC magnetic field with a high frequency of 10,000 Hz or more that penetrates the support for an effective time to induce a thermal rise of the magnetic fine particles, the reaction form of the fine particles spread on the reaction support is received by the infrared irradiation device. Since it is possible to read as a heat distribution using an apparatus, it is preferable in that a reaction form can be effectively identified even when a reaction support, a sample, a reagent, or the like contains a visible component.
As described above, when the reaction mode is developed on the side surface of a concave reaction support such as a cuvette or a test tube, a scanning type reading means such as a bar code reader is effectively used. Attempts are acceptable.

The signals read by the reading means correspond to drawings, images, etc. showing the respective forms of agglutination and non-aggregation obtained in each reaction using positive and negative samples as standards in advance in the comparison circuit. It is collated with the electric signal, and further, in the determination circuit, the presence or absence or the degree of aggregation is determined based on the comparison result from the comparison circuit. In some cases, the determination may be made by the operator directly on the screen or by performing arithmetic processing by an appropriate determination circuit. Further, the image, the light amount data and the judgment result read by the reading means are appropriately replaced by a tabulation table or a display symbol, and then stored by the storage means for a desired period of time, and are not displayed on the display screen or output paper. Is output.

In determining the reaction mode read by the reading means, the extent of the point-symmetric spread may be determined. The criterion at this time is applied to a reaction mode showing two or more different development lengths for each cohesion strength, starting from the point at which the strongest magnetic force acts on the reaction support.
Usually, it is roughly classified into two types: non-aggregated; (-), aggregated; (+). When the reaction form shows various development lengths or development densities in accordance with the cohesive strength, the maximum level of the development length may be appropriately divided and a plurality of criteria may be set in stages.
However, when a plurality of criteria are used, the difference in the development length between non-aggregation and aggregation is delicate, and for convenience, a criterion indicating determination suspension; Is also good.

The reaction conditions in the present invention can be experimentally determined in advance by the balance between the magnetic responsiveness of the fine particles and the magnetic force of the magnet, as long as the conditions described above are satisfied. Under the determined appropriate reaction conditions, the inclination angle of the pole face with respect to the reaction support and each development length level of the reaction form; (−), (±), (+), (++), (++)
+), Two typical trends are obtained as shown in Table 1 below. That is, (a) in the table shows the tendency when the separation distance between the magnet and the support for reaction is relatively short, and (b) in the table shows the tendency when the distance is relatively long. However, the tilt angle (θ) in the table refers to a tilt angle with respect to an imaginary plane such that a magnet that emits a magnetic flux in a vertical direction from the pole face is substantially parallel to the reaction support. The virtual plane here corresponds to a plane perpendicular to the shortest straight line connecting the reaction support and the magnet. An electric magnet or a plurality of different magnetic forces
When using a permanent magnet, it refers to an arrangement condition and a magnetic force condition such that the magnetic force distribution distributed on the same virtual plane shows a magnetic force gradient substantially equivalent to the magnetic force gradient provided by the single inclined magnet. .

[0044]

[Table 1]

As can be seen from Table 1, in the present invention,
The inclination angle formed by the magnetic support of the reaction support and the magnetic pole surface of the permanent magnet by the permanent magnet is such that each form level of the reaction form between the positive sample and the negative sample stably has a criterion difference of 1 or more. The effective range is 4 degrees or more and less than 50 degrees. Also,
Easiness of determination is an inclination angle between 4 degrees and less than 45 degrees that has a stable difference of 2 or more, and between 4 degrees and less than 34 degrees that has a difference of 3 or more, especially 11 degrees and less than 34 degrees It is preferable in terms of accuracy.

Hereinafter, a method and an apparatus for practicing the present invention will be exemplified. However, the present invention is not limited to these, and various combinations based on conventional techniques or prior knowledge obtained by a person skilled in the art. Design changes are possible to improve.

[0047]

Example 1 Preparation of Human IgG Immobilized Plate NUNC flat bottom module plate (2 × 8 well,
Individual wells (diameter 6 mm) were diluted with 0.01 M PBS, pH 7.4.
Human IgG (manufactured by Seikagaku Corporation) diluted to 15.6 μg / ml was added to 5
Dispense 0 μl / well and incubate at room temperature for 30 minutes. Then, it is washed with PBS and air-dried.

Preparation of Magnetic Particles Sensitized with Anti-Human IgG Antibody Fine particles containing magnetic material (average particle size: 6.1 μm, specific gravity: 1.2) were added to a 1% by weight solution in a 0.01 molar PBS solution of tannic acid (pH 7.4).
And incubate at 37 ° C for 30 minutes. The microparticles washed with the same PBS solution are incubated at 37 ° C. for 1 hour at 1% by weight in the same PBS solution containing 25 μg / ml goat anti-human IgG antibody (CAPPEL). After the antibody-sensitized particles are washed with the same PBS solution, the particles are washed with the same PBS solution containing 0.2% bovine serum albumin and 0.05% NaN _3.
Suspend to 0.25% by weight.

Preparation of Unsensitized Magnetic Fine Particles Unsensitized magnetic fine particles are prepared in the same manner as described above except that the concentration of the goat anti-human IgG antibody is 0 μg / ml.

Preparation of a deployment device A cylindrical ferrite magnet (diameter: 10 mm, magnetic flux density: approx.
2,500Gs) 125 is divided into two semi-circular magnets by the same procedure as described in FIG. 1, cut along the dotted line, and then adhered with a strong adhesive to form a substantially circular shape. A composite magnet 2 having a pole face was prepared. The adjacent angle between the adjacent north pole and south pole is 140 degrees.

Formation of Reaction Form Derived from Aggregation Reaction The prepared antibody-sensitized magnetic fine particles and unsensitized magnetic fine particles were respectively added to two wells of the prepared IgG-immobilized plate.
Add in aliquots and stir. After stirring, the plate is allowed to stand horizontally, and the top joint portion of the circular magnetic pole surface of the composite magnet prepared is positioned at the center of the lower surface of the flat bottom of the well, whereby the inclination angle of each magnetic pole surface with respect to the well bottom surface ( θ)
Were positioned at 20 degrees each. After applying the magnet,
The reaction form of the fine particles that have been developed within 3 minutes is photographed, and determined based on the above-described five-step criteria. As a result, the reaction pattern due to the sensitized particles has a distribution that spreads almost uniformly on the bottom of the well, whereas the reaction pattern due to the unsensitized particles forms a distribution gathered in the center of the well where the magnet is closest. I do. The reaction mode of the fine particles once formed on the flat bottom of the well is stably maintained for 7 days or more even after the magnet is removed. Further, in the above example, after the sensitized and unsensitized fine particles were added to each well and stirred, the disk-shaped magnet was approached parallel to the flat bottom well, so that all the fine particles were reduced to 30 regardless of the agglutination reaction. After evenly distributing the entire bottom of the well in about seconds, if it is inclined at 20 degrees, a reaction form that can be determined after 60 to 90 seconds is obtained.

In this embodiment, the results obtained by variously changing the inclination angle (θ) of the pole face with respect to the bottom of the well show the same tendency as (a) in Table 1 described above. On the other hand, when the top joint portion of the circular magnetic pole surface of the composite magnet was positioned at a distance of 5 to 10 mm from the center of the lower surface of the flat bottom of the well, the above Table 1 was used.
(B). Therefore, it is designed to always obtain the optimal reaction form by replacing it with a composite magnet adjusted to the optimum angle or adding a configuration that can adjust the angle according to the type of test object, dispensing volume, etc. can do.

[0053]

Example 2 Whole blood after blood collection was diluted to a predetermined concentration with physiological saline, and 1% by weight of magnetic particles were added to a cuvette (10 mm × 75 mm) having a square cross section shown in FIG. Is dispensed in a predetermined amount and incubated for a certain period of time, and 2 ml of a diluent is added to the cuvette after the agglutination reaction. A magnet having a pyramid-shaped quadrangular prism is formed by laminating a magnet having a cross section of a regular triangular prism so that the two magnetic pole surfaces 3a and 3b are adjacent to each other at a right angle. Face each other so as to intersect at right angles. A point-symmetric distribution pattern on the side surface of the cuvette formed after a predetermined time is read two-dimensionally by an imaging device positioned on the side opposite to the magnet. The read image signal is arithmetically processed by the arithmetic unit as a luminance distribution in the diameter direction passing through the center of the distribution form,
The reaction (positive) and the unreacted (negative) are discriminated by being compared with the luminance distribution based on the coagulated / non-coagulated image as a reference. In the above embodiment, it is needless to say that the cuvette can be used in the same manner even if the cuvette has a rectangular cross-section, and further, the cuvette can be changed in side surface angle as needed, or can be bent in multiple stages. May be used.

[0054]

[Embodiment 3] FIG. 10 is a perspective view showing a main part of an apparatus in which the method described in Embodiment 2 is implemented, and FIG.
FIG. 2 is a cross-sectional view of A ′ and a block diagram of each control unit. In a turret 50 that moves intermittently at a constant pitch, for example, every five minutes, a plurality of rectangular cuvettes 51 are housed on the circumference at equal intervals and freely removably. Where the cuvette 51
As shown in FIG. 10, the diagonal line of the cuvette 51 coincides with the diameter line. The rotation of the turret 50 is
By connecting to a rotation control unit (not shown) connected to an analysis command unit that makes it possible to change the rotation speed based on the information input by the data, control is performed according to a desired analysis item. Around the turret 50, each test sample to be analyzed,
For example, a sample container 52 containing diluted serum, whole blood, etc.
It is transported while being held at 52. The rotation of a sample dispenser 54 connected to a suction / discharge means (not shown) is controlled so as to intersect the transfer path of the sample container 52 and the transfer path of the cuvette 51 on the turret 50. A plurality of reagent containers 55 each containing a reagent corresponding to an analysis item are arranged on the circumference of a reagent table 56 whose rotation is selectively controlled. The reagent dispenser 57 connected to the suction / discharge means (not shown) includes a sample dispenser 54.
Are rotated and moved up and down between the cuvette 51b at the next stop position of the cuvette 51a that can intersect and the reagent container 55b at the predetermined stop position on the reagent table 56. Cuvette 51b crossing reagent dispenser 57
At the next stop position, a diluent dispenser 59 communicating with a diluent tank 58 containing a sufficient amount of diluent is positioned near and above the cuvette opening. The diluent dispenser 58 is controlled so that a predetermined amount of diluent is transported from the diluent tank 58 at regular intervals and supplied to the corresponding cuvette. Further, at the next stop position of the diluent dispenser 59, the prism magnet 60 created as described with reference to FIG. At this time, the orientation of the prismatic magnet 60 is fixedly arranged at an opposing position such that the joining line formed by the adjacent magnetic pole surfaces is orthogonal to the straight line of the side edge of the cuvette 51c at which the cuvette 51c stops. The separation distance between the cuvette 51c and the prism magnet 60 is adjusted so that an appropriate reaction time can be obtained from a range where substantially parallel magnetic fluxes emitted from the magnetic pole surfaces reach.

Here, as shown in FIG.
A disk 62 having a plurality of detection holes 61 provided at equal intervals on the circumference at the lower portion of the rotary shaft 50 '
The light source 63 and the light receiving element 64 are opposed to a fixed position above and below the detection hole 61, respectively. Light receiving element 64
Is connected to a position determination unit including an appropriate counting circuit, and the position determination unit is connected to the calculation unit and the rotation control unit, thereby enabling rotation control based on the stop position of the turret 50.
On the opposite side of the magnetic pole surface on which the prism magnet 60 is fixedly arranged, reading means 65 facing the cuvette 51c is provided.
For example, a CCD is positioned. This reading means 65
Is connected to a position determination unit and an appropriate display unit and / or storage unit via a calculation unit including a calculation circuit, a comparison circuit, a determination circuit, and the like (not shown) via a lead line.

At a stop position next to the stop position where the prism magnet 60 and the reading means 65 face each other, replacement means is arranged. The exchange means is a cuvette 51d on the turret 50.
And a gripper arm 67 for gripping the cuvette 51d in a lifted state by a spring force. The support rod 68 supporting the gripping arm 67 is a gripping arm.
The arm 67 and the lift member 66 are retracted from above the turret 50 to the outside of the outer periphery, and are connected to driving means (not shown) so that the arm 67 approaches the stop position of the corresponding cuvette 51d. A cuvette supply unit and a collection unit (not shown) are arranged outside the turret 50 on which the gripping arm 67 rotates,
At an appropriate rotation stop position of the arm 67, the used cuvette is pulled down to a predetermined collecting box (not shown), and the cuvette caught from the cuvette supply unit is pushed out toward the arm 67. It has a mechanism to make it grip.

According to the apparatus shown in FIGS. 10 and 11 having such a configuration, when the cuvette held by the turret sequentially stops at the sample dispensing position and the reagent dispensing position, the sample dispenser and the reagent dispenser are dispensed. Aspirates a required amount of a sample and a reagent from a desired sample container and a reagent container, respectively, and dispenses into a cuvette and mixes. After the mixing, the intermittent movement is repeated as it is for a predetermined time, and the aggregation reaction proceeds. The cuvette corresponding to the time when the agglutination reaction ends is stopped at the diluent supply position, and a certain amount of diluent is added,
The magnetic fine particles in the cuvette are suspended. The timing for adding the diluting liquid may be any time during the time when the diluting liquid is stopped at the diluting liquid dispensing position, but may be any timing convenient for developing the reaction mode by the next magnet. For example, if the sample contains blood cells with a high specific gravity, it is preferable to add the diluent sooner. In order to obtain a turbid state, the addition of the diluent may be terminated immediately before the cuvette moves to the next stop position. The cuvette to which the diluent has been added is subjected to the magnetism generated by the prism magnet at the next stop position. At this time, the magnetic fine particles in the cuvette move in accordance with the magnetic attraction that converges line-symmetrically and roll along the line-symmetrical inclined surface of the cuvette side surface. Is developed on the side surface of the cuvette.

The developed reaction form is read by the reading means facing the magnet, converted into an electric signal, and transmitted to the arithmetic unit. The arithmetic unit performs arithmetic processing on the read signal and determines the presence and / or degree of aggregation based on a predetermined determination criterion. The determined result is stored, displayed, and recorded as appropriate.
On the other hand, the cuvette read by the reading means moves to the next stop position, where it is replaced with a new cuvette. According to the present embodiment, by developing the reaction form on the side surface of the cuvette, a reading device for a plurality of conveyed reaction supports can be adopted as a measuring instrument used in the absorbance measurement. It is easy to double use for checking the properties of reagents and the dispensed amount, etc., and is excellent in economy and multifunctionality. In addition, the arrangement of the reading means can be efficiently accommodated in the apparatus as compared with the conventional arrangement in the vertical direction, which contributes to downsizing. Furthermore, since the reaction form can be developed in a direction different from gravity, even when the sample etc. contains sedimentation components such as blood cells, the reaction form is affected by deposits that obscure the reaction form. This is preferable in that it can be read without using a computer.

[0059]

Example 4 Two flat glass slides (10 mm × 10 mm) on which a predetermined antigen was uniformly coated were provided, and two glass slides were faced in parallel through a gap (0.5 mm) that would cause a capillary phenomenon. The slid reaction vessel 26 is set upright in the vertical direction to open the gap upward and downward. After a predetermined concentration of HBs-sensitized magnetic reagent solution is spotted and held in a fixed amount in the upper gap of the slide-shaped reaction vessel, one magnet having a flat surface of 10 mm × 10 mm as a magnetic pole is attached to the slide glass. The magnetic substance-containing reagent is uniformly distributed on the slide glass surface by sandwiching and facing each other in parallel from both sides. next,
While maintaining this state, whole blood of a predetermined dilution concentration is spotted and allowed to react at room temperature for a certain period of time, and then the diluent is spotted several times to sufficiently wash out blood components in the reaction vessel. A predetermined dilution of the anti-IgG antibody reagent is spotted, filled in the gap of the slide glass, and allowed to react at room temperature for a predetermined time.
By confronting the convex step magnet as shown in FIG. 5A, a concentric reaction mode is developed on the slide glass surface. Observe the distribution form after formation with the naked eye,
Identify reacted or unreacted.

[0060]

Embodiment 5 FIG. 7 shows the pole faces 25a, 25a of an electric magnet connected to a common power supply and a transformer (not shown).
b is formed into a circular shape that is parallel and equal in center, and the intersection of the diagonal lines of the pole faces 25a, 25b intersects the lower plate inner wall 26a of the reaction support consisting of the two upper and lower glass plates 26a, 26b. FIG. 2 shows a deploying device positioned so as to be focused on FIG. Here, the upper and lower glass plates 26
a, 26b are gaps that are extremely narrow enough to hold the liquid by capillary action,
For example, it is assumed that the flat surface is 0.1 to 1.0 mm and faces in parallel. When a reaction solution containing magnetic fine particles that have undergone an agglutination reaction with the test sample is spotted in the gap, the reaction solution is quickly introduced by capillary action, and a certain amount of the reaction solution is held. here,
The reaction solution can be introduced into the reaction support by simultaneously or sequentially applying the test sample and the reactive magnetic fine particles separately by separate application means, or immediately after mixing or after a certain period of time after mixing. It may be spotted by spotting means. The direction in which the circular pole tip faces should depend on the shape of the reaction support to be used, and should be substantially parallel to the orientation of the wall surface of the reaction support to form a distribution of magnetic particles. Let it. Here, the magnetic fine particles introduced into the glass plates 26a and 26b and subjected to the agglutination reaction with the predetermined sample are drawn concentrically along the inner wall 26a of the lower plate. Develop different reaction forms of aggregation.

FIG. 14 shows an analyzer to which the developing device of the fifth embodiment is applied. That is, the reaction support 81 in the form of a flat plate is accommodated at regular intervals around the circumference of the turret 80 which rotates intermittently at a predetermined pitch. It is assumed that a peptide antigen such as HIV has been coated on the reaction support 81 in advance. Techniques for purifying or synthesizing such antigens and coating techniques are well known and will not be described here. First, when the support exchanging device 91 pushes the support 93 in the standby state forward, it slides along the notch on the peripheral edge of the turret 80 and is mounted at a predetermined position. Next, after one pitch (for example, 3 to 10 minutes), the sample solution from the diluted sample container 82 is spotted by the sample dispenser 83 and is incubated for a predetermined time. After the reaction, the support 81 is washed by the washing device 84 by draining the sample solution to the waste liquid tank 85 and repeatedly supplying and draining the washing liquid (for example, PBS, physiological saline, etc.). After washing, the support 81 is sprayed with a reagent solution containing magnetic particles sensitized with anti-human IgG antibody as shown in Example 1 from a reagent container 87 by a reagent dispenser 88.
The dispenser of the sample and the reagent forms a thin tube that can move by capillary action, and when the tip of the dispenser touches the gap of the reaction support 81 or is located very close to the gap, the spontaneous dispenser can be used. It is advantageous for economy and miniaturization to move to the space in the support 81 to complete the filling of a predetermined amount of liquid. The support 81 onto which the reagent solution has been spotted, after a certain time,
It stops at a position facing the magnetic pole surface of the developing device 89 as shown in FIG. 7 (A), and a reaction form by the agglutination reaction is developed during the stopping time. The reaction form after the development is read at the next stop position by a reader 90 including a light source and light receiving means (not shown). The read reaction mode is determined in the same manner as in Example 1. The support 92 that has been analyzed in this way is transferred to the turret 80 by the support exchange device 91.
And is discarded in a collection box (not shown).

[0062]

Embodiment 6 FIG. 13 shows an embodiment in which the configuration using the ring-shaped magnet described with reference to FIG. 6B is modified. That is, below the microplate 74, there is provided a magnet table 75 on which needle-like magnets 77 (see Japanese Patent Application Laid-Open No. 2-24464) corresponding to the individual wells 73 are arranged, while above the individual wells 73. A configuration similar to that of Example 1 was implemented, except that a rubber magnet plate 78 provided with a circular opening 79 corresponding to was positioned sandwiching it in a sandwich shape. Here, the needle magnet 77 is an electric magnet composed of a predetermined power supply, a transformer, and a coil 76 so as to facilitate electrical control. According to this embodiment, not only the operation and effect according to the description of FIGS. 6A and 6B can be effectively obtained, but also it can be easily applied to an automatic analyzer such as Japanese Patent Publication No. 2-16875. It is preferred in that respect.

[0063]

[Embodiment 7] FIG. 12 shows a developing device in a reaction mode using a magnet in which a magnetic pole surface is inclined stepwise on a reaction support having a flat surface. That is, an appropriately diluted patient serum is dispensed into a microplate 69 having a flat bottomed cylindrical well with a diameter of 6 mm on which a predetermined antigen, for example, an HCV peptide antigen is uniformly coated, and the mixture is incubated for a predetermined time and diluted. Wash with liquid. Next, 25 μl of a reagent solution containing 1% by weight of anti-IgG antibody-sensitized magnetic particles (5 μm) is dispensed. At this time, a step-shaped magnet 71 in which a ring-shaped magnet having a diameter different every 1 mm and a disk-shaped magnet (diameter 1 mm) at the top are superimposed on a convex support 70 made of a conical non-magnetic material having an inclination angle of about 20 degrees. (1000 to 3000 Gs), the microplate 69 is placed. On the upper surface of the microtiter plate 69, a flat plastic magnet 72 having a weak magnetic force of about 5 to 1/10 of that of the step magnet 71 is also mounted substantially simultaneously. At this time, on the bottom surface of the well, a magnetic gradient of concentricity from the step-shaped magnet 71 is connected to the magnetic force of substantially uniform distribution of the plastic magnet 72, and gradually increases concentrically toward the center. As a result, a stabilized magnetic field is formed. In such a magnetic field, a stable reaction mode can be developed at all times. In this embodiment, the step-like magnet 71 is formed of a fibrous magnet (a mixture of a magnet and a fiber member) having a diameter of less than 1 mm and is fixed concentrically or spirally at a high density. The improvement is preferred in that a clearer and more precise reaction form can be developed.

Comparative Example FIG. 15A is a configuration diagram showing a state in which a non-aggregation reaction mode using magnetic fine particles is developed in the direction of gravity using a conventional developing apparatus, and FIG. It is the figure which looked at the reaction form from the top. That is, 50 μl of whole blood of a healthy human after blood collection was dispensed into a glass test tube (diameter 10 mm) 95 having a hemispherical bottom surface, and polymer magnetic microparticles (diameter 4.5 μm) sensitized with HBs antigen 20 ng / ml. 50 μl (particle concentration: 0.65%) was added at 37 ° C. while stirring every 10 minutes.
Incubated for 10 minutes. Next, a disk-shaped rare earth ferrite magnet (diameter of 10 m
m, 1800 Gs) 99 circular magnetic pole faces were placed and left for 5 minutes. After the application of the magnetic attraction, as shown in FIG. 15A, a part of the red blood cells 97 in the reaction solution 96 containing whole blood in the test tube 99 becomes similar to the non-aggregated magnetic fine particles 98, as shown in FIG. Test tubes already 99
Deposits on the bottom. For this reason, as shown in FIG. 15 (B), a shadow due to red blood cells deposited around the magnetic fine particles 98 showing a non-agglutination reaction mode is observed. Of course, such unwanted shadows due to red blood cells also occur in the reaction form with positive samples. Therefore, a positive (+) judgment is made in spite of being negative (false positive), or a negative (-) judgment is made in spite of being positive (+) (false positive) Therefore, the longer the application time of the magnetic attraction (for example, 5 to 10 minutes or more), the more erroneous determination may be made.

[0065]

Example 8 A test tube (diameter 10 mm × 75 mm) was used, and the configuration of the magnet was a columnar magnet 100 having a semi-circular shape (radius 5 mm) and a pole face formed in a longitudinal direction and having a pole face as a longitudinal direction.
An apparatus and an inspection process similar to those of the first embodiment are performed. That is, a reaction solution obtained by mixing a whole blood sample and a reagent solution containing magnetic fine particles.
To 96, after a predetermined reaction time, 1 to 3 ml of a high-concentration sucrose aqueous solution (20 to 50 w / w%) is added to obtain a reaction solution 96 having a high specific gravity.
Next, the lower end of the semi-cylindrical magnet 100 was adjusted to a height of 5 to 15 mm from the bottom surface of the test tube 95, and the semi-circular pole face was positioned so as to be orthogonal to the side surface of the test tube 95. The reaction pattern 98 of the healthy human blood obtained by such a configuration clearly appears in a button-like manner at a position corresponding to an orthogonal point after a lapse of 3 to 5 minutes using the semi-cylindrical magnet 100. Here, the red blood cells in the reaction liquid 96 having a high specific gravity become relatively light and float without being submerged at a relatively high water level of the reaction liquid 96. On the other hand, the thus obtained reaction form is received by a photoelectric tube 101 positioned on the opposite side of the semi-cylindrical magnet 100, as shown in FIG.
The result may be displayed on the display means after data processing of the light receiving signal by the CPU. In this embodiment, it is within the scope of design matter to variously change the combination of the side surface shape and the magnetic pole surface shape of the concave container as shown in FIG. 2 as necessary. Further, additives for obtaining a reaction solution 96 having a high specific gravity include:
In addition to high-concentration sucrose, a separating agent such as phycol solution may be used.

[0066]

Ninth Embodiment FIG. 17 is a schematic partial sectional view showing an example of a developing device using the step-like magnet shown in FIG.
That is, a magnetic pin 107 (diameter 0.4 mm, length 6 mm) made of a ferromagnetic member (eg, iron oxide, stainless steel, etc.) has a disk-shaped support member 106 (diameter 0.4 mm) arranged in a matrix on the cylindrical support 105. 7 mm), and are densely inserted at regular intervals (0.8 mm) so as to be slidable up and down. The upper end of the magnetic pin 107 is hemispherically rounded. On the other hand, the magnetic pin 10
7, a terminal plate 109 is attached to the cylindrical support 10 via a deformable elastic member (for example, sponge, porous rubber, etc.).
5 Fixed to the bottom. Here, inside the elastic member,
A conductive spring member having flexibility and being connected to the lower end of each magnetic pin 107 is helically embedded to form a coil, and is electrically connected to a magnetic force control mechanism via a terminal plate 109. ing. The magnetic force control mechanism includes a shape recognition circuit for recognizing the inclination angle of the reaction support with respect to the direction of gravity based on the height position of each magnetic pin 107, and a desired magnetic gradient in consideration of the recognized shape. An arithmetic circuit for determining the magnetic force of each magnetic pin 107 and a magnetic force distribution unit for distributing the magnetic force from an electric magnet (not shown) to each magnetic force calculated by the arithmetic circuit. When a concave well 110 (diameter 6 mm) having a conical bottom is placed on such a developing device, each magnetic pin 107 comes into contact with the back side wall of the bottom of the well and is pushed down, so that the arrangement along the wall shape of the reaction support is achieved. Become.
At this time, each magnetic pin 107 is connected to an electric magnet by a magnetic force control mechanism (not shown) so that the magnetic force gradually increases concentrically toward the center.

According to such a configuration, the agglutination reaction by the magnetic fine particles is always performed under a desired magnetic force gradient regardless of the shape of the reaction vessel, so that a stable reaction mode can be developed. According to this embodiment, each magnetic pole end surface can be made to coincide with the wall surface of the reaction support at least irrespective of the shape, so that the magnetic force of each magnet is prepared in advance so as to give a constant magnetic force gradient to each magnet. Or by electrically setting and connecting, a constant magnetic force gradient can always be applied to any wall shape. Furthermore, the inclination of the reaction support wall surface is calculated and detected from the height difference between the magnetic pins, and if it is flat, the predetermined magnetic force gradient is applied as it is; By adjusting the height or the amount of electricity of each magnetic pin so that the attractive force distribution is predetermined, a mechanism for correction can be added. If such control can be performed, desired sensitivity can be obtained for a plurality of reaction supports irrespective of the shape, and reactivity of various sensitivities can be obtained for the same reaction support. It can be easily applied to the analysis of Also,
If the magnetic force control mechanism is configured to enable time switching by appropriate control means, magnetic force such as canceling the application of magnetism or stopping the movement of fine particles except during the development of the reaction mode Maintaining the reaction morphology by providing the distribution on the support can be easily achieved.

[0068]

Embodiment 10 FIG. 18A shows an elastic bag 111 (for example,
17 except that the pressing member 114 is a magnet having a magnetic pole surface that generates a substantially uniform magnetic force. Have. The configuration of the magnetic fluid 112 may be appropriately selected from commercially available products. The pressing member 114 is configured to be able to move up and down inside the cylindrical support table 105 by operating an operator or a machine by a pressing means (for example, a spring, a syringe, etc.) not shown. In this embodiment, a template 113 having a shape (for example, a conical shape, a hemispherical shape, etc.) giving a preferable magnetic force gradient is exchangeably positioned directly above each disk-shaped support member 106. I have. Here, it is preferable that a plurality of template shapes of the template 113 are prepared as required, and the exchange operation is controlled by an appropriate exchange means. In this embodiment, a microtiter plate having a hemispherical well as the template 113 is inverted and placed horizontally. At this time, after the plate-like slide glass 115 is positioned horizontally above the template 113, the pressing member 114 performs an indirect aggregation reaction with the magnetic fine particles. The magnetic fine particles on the flat plate are
The condensed and non-coagulated reaction modes described above are developed by concentric magnetic force gradients generated near the upper surface.
As described above, according to the developing device using the exchangeable mold, even when a reaction support having various shapes is used, the corresponding most preferable template is prepared and the shape of the reaction support is prepared. Can be recognized by an identification mark (for example, a bar code) and identified by an appropriate identification means, whereby an automatic analysis can be performed flexibly.

[0069]

Embodiment 11 FIG. 18B shows an elastic member 111 filled with a magnetic fluid 112 in each opening of a support member 116 having a columnar opening (diameter: 5 mm) formed in a matrix.
But the buffer mat 118 (eg, synthetic rubber, sponge, etc.)
Is attached to a circular convex portion (diameter: 5 mm) of the magnet table 119 via the. Further, the magnet table 119 is
Is biased by a spring 117, and a circular convex portion is fitted. By placing the microplate 120 in which circular flat bottom wells (diameter 3 mm) are arranged in a matrix, the same operation and effect as in FIG. In this embodiment, the magnetic fluid 11 is provided inside the template 113.
2 is a deformable structure, which is preferable in that a uniform magnetic gradient can be formed even with a small reaction support.
Further, in FIGS. 8A and 8B, it goes without saying that an electric magnet can be used similarly.

The present invention is not limited to the above-described embodiments, and various design changes can be made without departing from the scope of the claims and the matters described in the detailed description.

[0071]

According to the present invention, the same development area can be used.
By applying a point-symmetric magnetic gradient to
Of deployment pattern by magnetically responsive fine particles in the region
Since the morphology can be improved, it is easy to distinguish between agglutination and non-aggregation, so that the determination accuracy is high.

[0072]

[0073]

[0074]

[0075]

[0076]

[Brief description of the drawings]

1A is a schematic diagram showing a composite magnet used in the present invention, FIG. 1B is a diagram for explaining a method of manufacturing the composite magnet of FIG. 1A,

FIG. 2 (A) is a view showing an arrangement for developing a reaction mode on a side surface of a square cuvette, and FIG. 2 (B) is a view for developing a reaction mode on a side surface of a cylindrical test tube. FIG. 2C is a view showing an example of arrangement, and FIG. 2C is a view showing another example of arrangement for developing a reaction mode on a side surface of a cylindrical test tube;

FIG. 3 (A) is a diagram showing a non-aggregated reaction mode developed on the side surface of a square cuvette, and FIG. 3 (B) is a diagram showing an aggregation reaction mode developed on a side surface of a square cuvette. Diagram,

FIG. 4 is a diagram showing a configuration of an apparatus of the present invention having moving means,

5 (A) is a diagram showing the device of the present invention having a concentric step, FIG. 5 (B) is a diagram showing the device of the present invention having a polygonal pyramid,

6A is a diagram for explaining the principle of another developing device of the present invention, FIG. 6B is a diagram showing a configuration of a developing device to which the principle of FIG. 6A is applied,

7 (A) is a diagram showing a configuration of another embodiment of the present invention, and FIG. 7 (B) is a diagram showing a non-aggregation reaction mode obtained by the apparatus of FIG. 7 (A). , FIG. 7 (C) shows FIG. 7 (A)
A diagram showing a reaction form in the case of causing aggregation obtained by the apparatus of

8A is a diagram illustrating a configuration of a modification of the present invention, FIG. 8B is a diagram illustrating a magnetic force gradient caused by each composite magnet according to the configuration of FIG. 8A,

FIG. 9 is a view showing another embodiment of the modification shown in FIG.

FIG. 10 is a diagram showing an automatic analyzer that implements the configurations of FIGS. 2 and 3;

11 is a sectional view taken along the line AA ′ of FIG. 10,

FIG. 12 is a diagram showing an apparatus using the magnetic generator of FIG. 5,

FIG. 13 is a diagram showing an example of implementing the configuration shown in FIG. 6 (B);

FIG. 14 is a diagram showing an automatic analyzer for a developing apparatus of the present invention using a plate-like reaction support,

FIG. 15 (A) is a diagram showing a configuration of a conventional developing device using a whole blood sample, and FIG. 15 (B) is a diagram showing a reaction mode including a whole blood sample obtained by a conventional technique. ,

FIG. 16 (A) is a diagram showing a configuration of a developing device of the present invention using a whole blood sample, and FIG. 16 (B) is a diagram showing a configuration of a reading means in a reaction mode by the device of the present invention. ,

FIG. 17 is a diagram showing an apparatus configuration of the present invention for adjusting a magnetic force gradient via a deformable pin;

FIG. 18 (A) is a view showing an apparatus configuration for obtaining a constant magnetic force gradient using a deformable pin, and FIG. 18 (B).
Is a diagram showing another configuration using a deformable magnetic fluid,

[Explanation of symbols]

 2, 7, 8 Magnet 4 Cuvette 6 Test tube 11 Rotating plate 14 Arc magnet 15, 16 Ring magnet 27, 28 Composite magnet 29a, 29b, 30a, 30b Magnetic pole surface 50, 80 Turret 71 Step magnet 77 Needle shape Magnet 107 Magnetic pin 108,111 Elastic member 112 Magnetic fluid 113 Template 114 Pressing member 119 Magnet table

──────────────────────────────────────────────────続 き Continued on the front page (58) Field surveyed (Int.Cl. ⁷ , DB name) G01N 33/53-33/579

Claims (6)

(57) [Claims] The present invention relates to a reaction mode in which the presence or absence or degree of a binding reaction between a test substance and fine particles is morphologically identified using a group of magnetically responsive fine particles on which a substance that specifically binds to the test substance is immobilized. in a development apparatus, it is curved to the flat surface and including the straight line has an area of only deploying the required amount of particulate
A reaction support having such a development region, and a magnetic generator having a magnetic flux direction inclined with respect to a vertical axis of the development region , wherein at least two of the magnetic generators are the reaction support. To the same deployment area that includes
And a point-symmetric magnetic force gradient . 2. A method for immobilizing a substance that specifically binds to a test substance.
The test substance and the fine particles
Form that morphologically identifies the presence or degree of the binding reaction
The required amount of fine particles in
Curved or flat surface with a straight area
Support for reaction having such a development region, and a magnetic generator
Comprising a linear portion of the reaction support.
At least two magnetic sources acting on one deployment area
The living body is held with different magnetic poles so that the magnetic flux is connected.
As a result, a magnetic field gradient symmetrical with respect to the deployment area is generated.
A reaction type deployment device characterized by being arranged as follows . 3. A reaction system which uses a group of magnetically responsive fine particles on which a substance that specifically binds to a test substance is immobilized to morphologically identify the presence or absence or degree of a binding reaction between the test substance and the fine particles. In the deployment device, the required amount of fine particles is deployed.
Curved or flat surface with a straight area
A reaction support having such a development region, a magnetic generator having a magnetic flux direction inclined with respect to a vertical axis of the development region , and a linear portion of the reaction support having the magnetic generator.
Point symmetrical rotation trajectory with respect to the same deployment area
And a moving means for moving the developing device along an inclined state . 4. A reaction support according to claim 1 , wherein said magnetic generators have different diameters.
Claims characterized in that it is arranged so as to sandwich the development area of
3. The developing device of the reaction mode according to 2 . 5. A method for immobilizing a substance that specifically binds to a test substance.
The test substance and the fine particles
Form that morphologically identifies the presence or degree of the binding reaction
The required amount of fine particles in
Curved or flat surface with a straight area
Support for reaction having such a development region, and a magnetic generator
Wherein the magnetic generator is located near the deployment area.
A magnetic member to be mounted and a magnetic member for magnetizing the magnetic member.
And these magnetic members and magnets
Point symmetric with respect to the same developing area of the reaction support
A developing device in a reaction mode, wherein the developing device is arranged so as to generate a magnetic force gradient . 6. A step of reacting a required amount of a group of magnetically responsive fine particles on which a substance that specifically binds to a test substance is immobilized, with a sample whose presence of the test substance is unknown, Moving the group magnetically toward a predetermined wall surface;
The fine particles moving to the wall surface are moved along the predetermined wall surface .
A step of developing the particles in the developing region, and determining whether or not there is a reaction with the test substance based on the form of the expanded fine particles.
Other and a step of identifying the extent, the deployment step includes a step of forming a point-symmetrical magnetic gradient for the same expansion area of the wall, unreacted fine particles reaching the wall surface Is developed toward the point of symmetry of the magnetic force gradient.