JPH0643726Y2 - Immunological agglutination detector - Google Patents

Description

[Detailed Description of the Invention] [Industrial field of application] The present invention relates to an immunological agglutination detection device, and in particular, it can detect various blood groups from the agglutination pattern of blood cells and detect antigens and antibodies. An immunological agglutination detection device suitable for use.

[Conventional technology]

Conventionally, in the medical field, the agglutination pattern of blood cells, latex particles, and carbon particles is discriminated, and various components (eg, blood group, various antibodies, various proteins, etc.) in blood cells and viruses are detected and analyzed. Things are widespread.

Many immunological agglutination detection devices for detecting this kind of particle agglutination pattern have been researched and developed more than before and put to practical use. Examples of this immunological agglutination detection apparatus include those disclosed in Japanese Utility Model Publication No. 61-45479, Japanese Patent Publication No. 61-8934, and Japanese Patent Application Publication No. 59-98708.

[Problems to be solved by the device]

However, in the conventional example described in Japanese Utility Model Publication No. 61-45479, an image formed on a conical inclined bottom surface of a reaction container illuminated by a point light source is projected onto an image forming surface by a lens. Then, in the case where the light receiving element formed on this image plane and receiving the image scanned by the scanning device and converting it into an electric signal with time along the scanning direction according to the intensity of light is non-aggregated, the reaction container Since it has an entrance aperture that is approximately equal to or smaller than the image of the agglomeration pattern formed in the center of the bottom surface of the reaction vessel, the scanning line is at the bottom of the reaction vessel (where the agglomerates gather). There is a disadvantage that a structure is inevitably complicated because a positioning mechanism or the like is required to pass through. Further, since the slit forming the entrance opening of the light receiving element changes the agglutination pattern after the reaction depending on the type of immunological agglutination reaction as an inspection item, the opening area, shape, etc. must be adjusted, which is troublesome. There was an inconvenience.

Japanese Patent Publication No. Sho 61-8934 and Japanese Unexamined Patent Publication No. Sho 59-98.
In the one described in Japanese Patent No. 709, a collimator lens is used as an illumination lens for converting light emitted from a fixed point light source into a parallel light flux, and the parallel light is reacted through a diffuser plate (diffuser plate). The microplate in which the container is formed is uniformly illuminated, and the image of the conical bottom surface of each reaction container is imaged on the light receiving surface of the moving light receiving element by the imaging lens. Therefore, the relative positioning of the light source and the light receiving element must be performed with high accuracy, and there is a problem of accuracy of the collimator lens and an increase in size of the illuminating part. Furthermore, the collimator lens is very expensive. There is a disadvantage that the minute device becomes expensive.

[Purpose of device]

An object of the present invention is to provide an immunological agglutination reaction detection device which can improve the disadvantages of the conventional example and can reduce the cost and downsize the device without reducing the reliability of the detection result. To do.

[Means for Solving the Problems]

In the present invention, a multiplicity of reaction vessels having at least a part of the bottom surface inclined to form a matrix on a substrate are arranged in a matrix, and the agglutination reaction inspection plate is arranged on one side through the agglutination reaction inspection plate. And a light receiving section arranged on the other side. Then, each image of the agglomeration pattern formed on the bottom surface of the plurality of reaction vessels by the irradiation light output from the light emitting section is imaged through the lens on the light receiving surface of the light receiving element constituting the light receiving section, The agglutination pattern is detected by the output signal. Further, the light-emitting part is constituted by at least one point light source facing each reaction container for each reaction container forming an arbitrary column or row among a large number of reaction containers, and these The configuration is such that auxiliary point light sources are arranged at both ends of the row formed by the point light sources.
This is intended to achieve the above-mentioned object.

[Example of device]

In this example, human AB was used as an example of immunological agglutination.
Take the O type blood group determination test as an example.

Generally, when humans are classified by ABO blood type, all humans can be classified into four types, A type, B type, AB type and O type.

In order to distinguish each blood group by this blood grouping test, usually, blood collected from a subject is first centrifuged to separate it into red blood cells and serum.

When erythrocytes and serum of each of the above four blood types are mixed, as shown in Table 1 below, an aggregation phenomenon in which erythrocytes and serum stick to each other partially occurs. This is intended to discriminate each of the above blood types.

Here, in Table 1 above, x indicates non-aggregation reaction, and ∘ indicates aggregation reaction.

As can be seen from Table 1 above, O type red blood cells are AB type, B type, A type
Unlike the properties of type B red blood cells, type AB red blood cells are type A, type B
It can be said that they also have the properties of red blood cells of each type.

In this example, two sample liquids were prepared by injecting a diluent into each erythrocyte of each blood type, and anti-A serum (type B serum) and anti-B serum (type A serum) were added dropwise to each as a determination liquid. The blood type of the sample is determined by the method described above.

In this case, the blood type of the subject was A type and was agglutinated by the addition of anti-A serum, but was not agglutinated by anti-B serum. The sample blood was A type and was not agglutinated by anti-A serum but anti-B serum. The thing aggregated by is a B type. It can be determined that the one aggregated with both anti-A and anti-B serum was AB type, and the one not aggregated with both anti-A and anti-B serum was O type.

An embodiment of the present invention will be described below with reference to FIGS. 1 to 10.

In the embodiment shown in FIG. 1, a large number of reaction vessels 1 each having a conical bottom surface are arranged in a matrix as shown in FIG. 2 (see FIGS. 4 and 5). Microplate 3 as an agglutination test plate
And a light-receiving portion which is disposed on one side (upper side in FIG. 1) through the microplate 3 and a light-receiving portion which is disposed on the other axis (lower side in FIG. 1). Unit 10 and.

The light-emitting part 4 is an arbitrary column (here, the column i is referred to as appropriate) and a column adjacent to the column among a large number of reaction vessels 1, 1, ... Arranged in a matrix on the microplate 3 described above. Light emission as point light sources provided facing the reaction vessels 1 _1i , 1 _2i , 1 _3i , 1 _4i , 1 _5j , 1 _6j , 1 _7j , 1 _8j that form (here, j columns for convenience). Diode 2D, 2C, 2
It is composed of B, 2A, 2D ', 2C', 2B ', 2A'. Furthermore,
In this embodiment, both ends of each row formed by the light emitting diodes 2A, 2B, ..., That is, the light emitting diodes 2A, 2D, 2A ', 2
Light emitting diodes 2E, 2F, 2 as auxiliary point light sources outside D '
G and 2H are provided. Diffusing plates 31A and 31B are arranged between the light emitting section 4 and the microplate 3 in parallel with the microplate 3 at a constant interval. For this reason, in this embodiment, it is possible to effectively prevent a shortage of light quantity from occurring at both ends of each row formed by the light emitting diodes 2A.2B ....., and to irradiate the microplate 3 with substantially uniform parallel light. It has a structure that enables it.

On the other hand, as shown in FIG. 2, the light receiving unit 10 includes a lens holder 5 equipped with condenser lenses 6 at regular intervals so as to be able to face each reaction vessel 1, 1 ,. It is composed of a one-dimensional CCD sensor 7 held at the bottom and the condenser lenses 6, 6 ,.

More specifically, the lens holder 5 actually has the appearance shown in FIG. 3, and has a plurality of holes at intervals equal to the distance between the reaction vessels 1 and 1 adjacent to each other along the longitudinal direction. (4 in this embodiment) 5a, 5a, ...
Each condenser lens 6 is fixed to the peripheral wall of each hole 5a. At the bottom of this lens holder 5, the one-dimensional
A CCD sensor 7 is held below the condenser lens 6 at a fixed distance, that is, substantially the same distance as the focal length of the condenser lens 6, and parallel to the microplate 3. Therefore, for example, in FIG.
By the irradiation light from A, 2B, ..
Each image of the agglomeration pattern formed on the bottom surface of the four reaction vessels 1 _1i , 1 _2i , 1 _3i , 1 _4i arranged in a matrix and arranged in a matrix on the primary An image is formed on the original CCD sensor 7, and four reaction vessels 1
The image of the agglomeration pattern formed on the bottom surface of _1i , 1 _2i , 1 _3i , 1 _4i has a structure that can be detected at one time by the one-dimensional CCD sensor 7. Further, in this case, since each condenser lens 6 is held in each hole 5a formed in the lens holder 5, it is hardly affected by the light transmitted through the adjacent reaction vessel 1. It is like this.

Two light receiving units 10 are provided in this embodiment,
The reaction vessels 1 _1i , 1 _2i , 1 _3i , 1 _4i , 1 _5j , 1 _6j , in the i-th row and the j-th row are overlapped with each other on the upper surface of a movable plate 16 which will be described later. They are disposed respectively along the 1 _7j, 1 _8j. These light receiving units 10 and 10 are actually connected by a connecting member 10A shown in FIGS. In this case, the light receiving units 10 and 10 are arranged as shown in FIG. 4, and four holes 5a and 5a are formed at intervals equal to the distance between the reaction vessels adjacent to each other in the longitudinal direction. , ……, but the reaction vessels 1 _1i , 1 _2i , 1 _3i , 1 _4i , 1
It matches _5j , 1 _6j , 1 _7j , 1 _8j . In this case, the reaction vessels 1 are actually formed on the microplate 3 in a matrix of 8 rows and 12 columns, and the translucent light forming a part of the immunological agglutination reaction detection apparatus 20 shown in FIG. It is used by being placed on the horizontal plate 11 made of a flexible member.

The immunological agglutination reaction detection device 20 includes a horizontal plate 11, one support member 12A that supports the horizontal plate 11 from below, and the other support member 12B. Of these, support members 12A, 12B
A reinforcing plate 12C for connecting and fixing both of them is installed between the two. A guide shaft 13 is installed between the support members 12A and 12B along the longitudinal direction of the horizontal plate 11 as shown in FIG. Further, between the support members 12A and 12B, another shaft 14 having a male screw of a ball screw formed over its entire length is arranged parallel to the guide shaft 13 and rotatably equipped.

On the other hand, on both shafts 13 and 14, a box 15 shown in FIGS. 6 to 7 is equipped so as to be capable of reciprocating along the shafts 13 and 14. Specifically, the box
The hole 15 is provided with a hole 15a having a diameter substantially the same as the diameter of the shaft 13 and a hole 15b having a diameter substantially the same as the diameter of the shaft 14. Further, inside the box 15, a female screw portion of a ball screw in which a female screw (not shown) is formed which is opposed to the above-mentioned male screw via a ball (not shown) is built in.

On the upper surface of the box 15, a movable plate 16 for mounting the above-described light receiving unit 10 is arranged in parallel with the horizontal plate 11 and fixed. On the upper surface of the movable plate 16, support plates 18A and 18B for supporting the upper plate 17 having the above-mentioned light emitting diodes 2A, 2B, ...
Fixed orthogonal to 6. On the lower surface of the upper plate 17,
The diffuser plates 31A and 31B described above are integrally held. On the lower surface of the upper plate 17, the LED driver circuit (not shown) for driving the above-mentioned light emitting diodes 2A, 2B, ...
Is provided. Further, in this embodiment, the LED driver circuit is provided with light quantity adjusting means 22A to 22D for adjusting each light emitting diode in the relationship shown in FIG.

A substrate 19 arranged in parallel with the movable plate 16 is fixed to the upper surface of the movable plate 16.

This substrate 19 has a one-dimensional CCD sensor 7 composed of an IC or the like.
A CCD driver circuit (not shown) for driving is mounted.

Outside the support member 12A, a motor 21 that applies a rotational force to the shaft 14 through a gear mechanism (not shown) is installed. Therefore, in the present embodiment, when the motor 21 is driven, the movable plate 16 and the upper plate 17 integrally move the sixth plate while sandwiching the horizontal plate 11 and the micro plate 3 from above and below.
In the direction of arrow A in the figure, that is, along the row of reaction vessels 1, 1, ...

The one-dimensional CCD sensor 7 is a general-purpose one-dimensional CCD sensor in this embodiment.
CD sensor is used. The one-dimensional CCD sensor 7
Has a plurality of photoelectric conversion elements as photosensors arranged in a line on the light-receiving surface thereof, whereby the reaction vessels 1, 1, ...
The image of the agglomeration pattern formed on the bottom surface of the ... Is subdivided by the plurality of photoelectric conversion elements, and is converted into an electric signal from each photoelectric conversion element according to the intensity of light. In this embodiment, this electric signal is sent to a CPU (not shown) via an A / D converter (not shown), and the CPU determines the aggregation pattern.

Next, the operation of the immunological agglutination reaction detection device 20 configured as described above will be described.

First, prior to the detection of the aggregation pattern, the light amount adjustment means 22A, 22B, ... Specifically, for example, in the state of the first and fourth diagram, light emitting diodes 2A, and the reaction vessel 1 _4i, 1 _5j portion opposed to the 2D 'is brighter than the other (only the portion of three light-emitting diodes Since it receives light), the light amount of the light emitting diodes 2A, 2D 'is reduced, and at the same time, the light amount of the light emitting diodes 2E, 2H as auxiliary light sources is reduced. Next, the light emitting diodes 2B, 2C, 2
The light quantities of D, 2A ', 2B', and 2C 'are adjusted to this portion at the same illuminance, and the light emitting diodes 2F and 2G as auxiliary light sources are adjusted accordingly.

When the motor 21 is driven, the movable plate 16 starts to move, the positioning means (not shown) is controlled by the CPU (not shown), and the light receiving units 10, 10 shown in FIGS. When it is set to move down any column of 1, 1, ..., Light from the light emitting diodes 2A, 2B ,.
The microplate 3 is irradiated with light through A and 31B, and the light receiving unit 10 is irradiated by the light emitted from the light emitting diodes 2A, 2B, ....
A total of eight reaction vessels located above the
1 _1i , 1 _2i , 1 _3i , 1 _4i , 1 _5j , 1 _6j , 1 _7j , 1 _8j ...... Each image of the agglomeration pattern formed on the bottom surface of one-dimensional CCD through each condensing lens 6. An image is formed on the sensors 7, 7. The output signal from the one-dimensional CCD sensor 7, 7 is sent to a CPU (not shown) via an A / D converter (not shown), and in the CPU, the movement amount of the movable plate 16 is changed from the feed amount (rotation amount) of the motor. It is designed to calculate which column of the reaction vessels is being inspected and automatically determine the agglutination pattern of the analyte in each reaction vessel.

Hereinafter, an example of the determination in this case will be described.

In the above ABO blood type determination method, when an agglutination reaction occurs, a mass of blood cell particles bound to each other via serum is uniformly deposited on the conical bottom surface of the reaction vessel 1 like snow. When the agglutination reaction does not occur, the blood cell particles settle down in a discrete manner without interposing serum, reach the conical bottom surface, roll down the slope, and collect and accumulate at the center of the bottom surface.

FIG. 10 is an enlarged view of the bottom surface of the reaction container 1.

This shows a state in which the radius of the bottom surface of the reaction container 1 is 6 mm, the depth of the inclined portion is 1.5 mm, and the inclination angle is 30 °, and the particles are aggregated and uniformly deposited on the conical bottom surface. ing.
Such a uniform deposition pattern is, for example, in the ABO blood typing test, anti-A
Obtained when serum (type B serum) is added and spontaneously precipitated. That is, in this case, since red blood cells are bound to each other by serum, they do not roll down the inclined surface and are almost uniformly deposited on the bottom surface. When this uniform deposition pattern is observed in detail, it is fairly thick at the lowermost point A in the center, whereas it is slightly thinner at the peripheral point C, and the intermediate portion B between them is slightly thicker. At the point, the thickness changes almost continuously. In this case, the light transmission amount has a minimum value at the point A, gradually increases from the point A toward the peripheral portion, and reaches a maximum near the point C. Because of this, one-dimensional
The output of the CCD sensor 7 changes correspondingly, so the OPU
Then, the uniform deposition pattern is determined (here, the blood of the sample is A type).

As described above, according to this embodiment, as shown in FIGS. 3 to 4, the light receiving units 10 are connected by the connecting member 10A in such a state that the two light receiving units 10 partially overlap each other in the longitudinal direction. The one-dimensional CCD sensor 7 formed in a so-called crank shape and mounted on the bottom of each light receiving unit 10 is arranged along the vertical column of the reaction vessels 1, 1, ... arranged in a matrix on the microplate 3. To detect 8 images of the agglomeration pattern formed at the bottom of the reaction vessels 1, 1, ... Can be done. Therefore, the agglutination reaction of the specimens in all the reactors 1 of the microplate 3 can be detected only by scanning in one direction,
There is an advantage that the positioning accuracy can be improved and the inspection time can be remarkably shortened as compared with the case of two-direction vertical and horizontal scanning. Further, the light emitting diodes 2E, 2F, 2G, 2H as auxiliary point light sources arranged at both ends of the row formed by the light emitting diodes 2A, 2B, ... There is an advantage that the light quantity can be made uniform very easily, and since the illumination lens is not used, the cost can be reduced accordingly and the apparatus can be downsized. Further, since a light emitting diode is used as a light source, it has advantages that reliability and stability can be improved and power consumption can be reduced.

The light quantity adjusting means 22C may be replaced with light quantity adjusting means 22E, 22F, 22G as shown in FIG. 9, and the adjusting function of the light quantity adjusting means 22C may be divided into three parts. In this case, finer adjustment is possible.

[Effect of device]

Since the present invention is configured and functions as described above, according to the function of the auxiliary point light source, the agglutination reaction test can be performed by using the diffuser plate as in the above embodiment without using a lens for illumination. It is possible to irradiate each reaction container formed on the application plate with light of uniform illuminance.
For this reason, the illumination lens such as the collimator lens can be omitted without reducing the reliability of the inspection result, and the optical system can be simplified accordingly and the apparatus can be downsized and the cost can be reduced. It is possible to provide a practical immunological agglutination detection device that is not possible in the past.

[Brief description of drawings]

FIG. 1 is a block diagram showing the main part of one embodiment of the present invention, and FIG.
The figure is an explanatory view showing the internal structure of the light emitting part and the light receiving part of FIG.
3 is an external perspective view of the light receiving unit of FIG. 1, FIG. 4 is an explanatory view showing an example of the arrangement of the light receiving unit of FIG. 1, and FIG.
FIG. 7 is an overall view showing the microplate of FIG. 1, FIG. 6 is an external perspective view showing the entire embodiment of FIG. 1, and FIG.
FIG. 8 is a diagram showing a state seen along the line III-III in FIG. 8, FIG. 8 is an explanatory diagram showing the relationship between the light amount adjusting means and the light source in the embodiment of FIG. 1, and FIG. 9 is a diagram showing other relationships. , FIG. 10 is a diagram for explaining the operation of the embodiment shown in FIG. 1 ... Reaction container, 2 ... Substrate, 2A, 2B, 2C, 2D ... Light emitting diode as point light source, 2E, 2F, 2G, 2H ... Light emitting diode as auxiliary light source, 3 ... For agglutination reaction inspection Micro plate as a plate, 4 ... Light emitting unit, 10 ... Light receiving unit as light receiving unit.

Claims (1)

[Scope of utility model registration request] 1. An agglutination reaction inspection plate in which a large number of reaction vessels having at least a part of the bottom surface inclined are formed in a matrix on a substrate, and the agglutination reaction inspection plate is arranged on one side through the agglutination reaction inspection plate. And a light-receiving unit disposed on the other side, and the images of the agglomeration patterns formed on the bottom surfaces of the reaction vessels by the irradiation light output from the light-emitting unit are formed on the lens. In the immunological agglutination reaction detection device, which forms an image on the light receiving surface of the light receiving element that constitutes the light receiving portion via the output signal and detects the agglutination pattern by the output signal, , Each of the reaction vessels forming any column or row is constituted by at least one point light source facing each reaction vessel, and both ends of the row formed by these point light sources Immunological agglutination reaction detecting apparatus characterized in that a supplementary point light source.