JPH0422844A - Immune reaction component detector - Google Patents

Abstract

PURPOSE:To detect immune reaction components with simple constitution by forming the photodetecting part of a non-dimensional linear image sensor as an imposing part for a reaction liquid. CONSTITUTION:Photosensitive elements 3a are disposed in one array and a transparent glass plate 3b is provided atop the elements to form the one- dimensional CCD image sensor 3. The sensor is disposed atop a body case 2 of the immune reaction component detector 1. Aggregate is formed in the reaction liquid L subjected to an antigen-antibody reaction on cover glass G and the quantity of the received light detected by the elements 3a right thereunder decreases when the reaction liquid L is imposed on the photodetecting part of the sensor 3. Namely, the quantity of the received light of the respective elements 3a changes in accordance with the aggregation state and the number of the aggregate of the sensitized carrier particles is nearly proportional to the concn. of the immune reaction components in he specimen and, therefore, the immune reaction components can be detected, i.e. the qualitative and quantitative detection of the antigen-antibody reaction is executed by counting the number of the picture elements to be shielded of light by the agglutinate.

Description

Detailed Description of the Invention [Field of Industrial Application] The present invention relates to an apparatus for detecting immune reaction components, and more specifically, the present invention relates to an apparatus for detecting an immune reaction component, and more specifically, a method for detecting an immunoreactive component between a sample and a sensitized carrier particle in which the carrier particle is sensitized with an antibody or an antigen. The present invention relates to an immune reaction component detection device that quantitatively or qualitatively detects the presence of antibodies or antigens in a specimen by measuring the degree of aggregation of sensitized carrier particles in a reaction solution.

[Prior art] Conventionally, sensitized latex, which is obtained by sensitizing latex with antibodies or antigens, is reacted with a specimen in a liquid medium, and the reaction solution is observed with the naked eye to confirm the agglutination state of the sensitized latex. This enables qualitative detection of immune reaction components. In addition, as an immune reaction component detection device, the above reaction solution is injected into a transparent test container, mounted on a predetermined holding device, and the test container is irradiated with a light beam such as a laser (2). It is also known to quantify the state of aggregation by forming an image with a lens and measuring the intensity distribution with a phototube or the like.

[Problems to be Solved by the Invention] However, in the above-mentioned device, the optical system mechanism from the light source to the light receiving device such as the phototube becomes extremely complicated, resulting in a large-scale configuration. That is, a light source such as a laser, a lens to uniformly irradiate the test container with light, a holding device to hold the test container in a predetermined position relative to the light, and a lens to image the diffracted light. etc. These mutual positional relationships must also be arranged with precision. Also, instead of a phototube, a TV
Although it is known that a camera is used, even in this case, problems such as the complexity of the optical system mechanism and the accuracy required for the arrangement of each mechanism remain unsolved.

The objective of the immune reaction component detection device of the present invention is to solve the above-mentioned problems and perform qualitative or regular examination of immune reaction components with a simple configuration.

The structure of the present invention that achieves the above object will be explained below.

[Means for Solving the Problems] The immune reaction component detection device of the present invention detects the degree of aggregation of sensitized carrier particles in a reaction solution of a sample and sensitized carrier particles in which carrier particles are sensitized with antibodies or antigens. In an immune reaction component detection device that quantitatively or qualitatively detects the presence of antigens or antibodies in the specimen by measuring, a plurality of photosensitive elements are arranged in one dimension and the light receiving part is placed on which the reaction solution is placed. The present invention is characterized in that it comprises: a linear image sensor as a part; and a calculation means for calculating the degree of aggregation of the sensitized carrier particles based on the output value of the linear image sensor.

[Function] The immune reaction component detection device of the present invention having the above-mentioned configuration is equipped with a linear image sensor in which a plurality of photosensitive elements are arranged in one dimension, and a reaction solution is placed on the light receiving part. When aggregates are formed, the amount of light detected by the photosensitive element directly below them decreases.In other words, since the light-receiving part of the linear image sensor is used as a mounting part for the reaction solution, it is necessary to use a special light source, lens, etc. Even if it is not used, the amount of light received by each photosensitive element changes depending on the state of aggregation.The number of aggregates of sensitized carrier particles in the reaction solution is approximately proportional to the concentration of the immune reaction component in the specimen, and The number of aggregates distributed on a two-dimensional plane is 1
Since the number of aggregates dispersed in a dimension is proportional to 2, the degree of aggregation of the sensitized carrier particles calculated by the calculation means based on the output value of the linear image sensor 1 is the quantitative or qualitative value of the antigen or antibody in the sample. Respond to detection.

Note that placing the reaction solution includes placing it on the light receiving section of the linear image sensor through a transparent body such as a cover glass.

[Example] In order to further clarify the structure and operation of the present invention described above, preferred examples of the immune reaction component detection device of the present invention will be described below.

FIG. 1 is a schematic perspective view of the immune reaction component detection device of the present invention, and FIG. 2 is a schematic configuration diagram thereof.

Immune reaction component detection device (hereinafter simply referred to as detection device) 1
is a CCD image sensor 3. Sensor driver 5. Level conversion amplifier B Comparator 9゜Level adjustment knob 1
], counter 13. D/A converter]5. LCD display 17
2 display driver 19 and the like. As shown in FIG. 3, the CCD image sensor 3 is a well-known one-dimensional CCD in which photosensitive elements 38 using photodiodes are arranged in a row in the center.
This is an image sensor, and in this embodiment, the number of photosensitive elements 3a (number of pixels) is 2048, and the size of the photosensitive surface of each photosensitive element 3a is 14×14 μm. Also, CC
A transparent glass plate 3b is provided on the upper surface (light receiving section) of the D-image sensor 3, and as shown in FIG. 1, it is disposed on the same plane as the upper surface of the main body case 21 of the detection device.

The sensor driver 5 includes an oscillation circuit, a timing control circuit, etc., and drives the CCD image sensor 3. When the CCD image sensor 3 is driven, a time-series signal corresponding to the amount of light received by each pixel is output, amplified by the level conversion amplifier 7, and input to the comparator 9. The comparator 9 compares the output value (after amplification) for each pixel in scanning repeated at a predetermined period with a predetermined threshold value vO as shown in FIG. 4(b).
As shown in FIG. 2, a binarized video signal (time series signal) that becomes a high level signal when the output value is less than the threshold value VD is output to the counter 13. This threshold value vO is
Level adjustment knob 11 provided on the top surface of the main body case 21
It can be changed by operating.

The counter 13 counts the number of high-level pulses in each scan from the input video signal. That is, the number of pixels forming a dark area (or a bright area) obtained by comparing the brightness of each pixel with respect to a predetermined level is counted. Since the number of pixels of the CCD image sensor 3 is 2048, the value of the counter 13 takes a value in the range of 0 to 2048. The output of the counter 13 is input to the D/A converter 15, which outputs a voltage in the range of -2V to 2V according to the counter value. In other words, if the counter value is 0, the voltage is 2V.
, If the counter value is 2048, it is set to -2V, and this range is divided into 2048 steps to output a voltage according to the counter value. This output signal is supplied to an output terminal 23 provided on the main body case 21. The output terminal 23 is used to connect a measurement device such as an oscilloscope or a high coder to measure detailed output waveforms and the like. Also, D/A
The output of the converter] 5 is also input to the display driver 19. The display driver 19 drives the 7-segment liquid crystal display 17 provided on the main body case 21, converts the output voltage from the D/A converter 15 into a digital signal, and displays the voltage value on the liquid crystal display. display on the device 17. Therefore,
The liquid crystal display 17 digitally displays values in the range of -2V to 2V.

As shown in FIG. 1, the testing device 1 described above sends a reaction liquid obtained by causing an antigen-antibody reaction on a transparent body such as a cover glass G to the light receiving part of the CCD image sensor 3 (glass plate 3b).
) to perform immobilization and quantitative detection of antigens or antibodies. In this example, the measurement target was CRP.
(C-Reactve protein).

In addition, as measurement reagents (Table: CRP sensitized latex solution in which latex was sensitized with anti-CRP antibody and phosphate buffer (p)-17,2), and CRP standard solution as a specimen (
1.18 mg/d I) was used.

In addition, the CRP sensitized latex solution and phosphate buffer are as follows:
01 Latex agglutination reagent for CRP assay manufactured by Medical and Biological Research Institute (CRP-E kit, detection sensitivity o,
.

7mg/c11).

The CRP-sensitized latex particles of the CRP-sensitized latex solution have a diameter of 0.1 to 1. 0 μm and reacts with CRP to form larger aggregates (approximately 50 μm in diameter).

The size of each pixel of the COD image sensor is 14x14μ
m, the CRP-sensitized latex particles are CCD
Although each photosensitive element 3a of the image sensor 3 does not block the insulting light, when aggregates are formed, the light is blocked and the output of each photosensitive element 3a decreases. The number of aggregates is proportional to the concentration of the analyte (CRP), and the number of aggregates uniformly dispersed on a two-dimensional plane is proportional to the number of aggregates dispersed on one dimension. For this reason, the detection device of this embodiment] detects the concentration of the sample by counting the number of pixels that become dark areas blocked by direct light. , using normal indoor lighting.

Next, a method for measuring an antigen-antibody reaction using the apparatus of this example and its results will be explained.

First, a 50 μm CRP standard solution is dropped onto the cover glass G, and then a CRP sensitized latex solution is dropped onto the cover glass G. While stirring for 2 minutes with a suitable stirring rod or the like, the reaction liquid is spread over a glass cover glass and placed on the top surface of the CCD image sensor 3, as shown in FIG. At this time, the CCD image sensor 3 is placed so that the reaction liquid covers the entire photosensitive surface. Subsequently, the value displayed on the liquid crystal display 17 is read.

Similarly, change the concentration of the CRP standard solution (dilute up to 32 times) and repeat the above operation.

FIG. 5 is a graph showing measurement results in which output values (voltages) read for each concentration are plotted. The measured value for each concentration fluctuates with a slight range because it is a digital display, but the average value of this fluctuation range A (indicated by ○J mark) shows almost linear characteristics up to 8 times dilution. Naf-,
In the case of 16-fold dilution, the concentration of CRP is 0.0737 m
g/d l (1°18/16=0.0737), and the detection sensitivity of the latex agglutination reagent for CRP quantification (0,
07 mg/d1), it can be said that this measurement result accurately detects the concentration of CRP. That is, the output value obtained by measurement can be regarded as a function of the concentration of CRP. This confirms that the number of aggregates is proportional to the concentration of the analyte (CRP), and the number of aggregates uniformly distributed on a two-dimensional plane is proportional to the number of aggregates distributed on one dimension. ing. Moreover, it can be said that the diffusion of light between the aggregate and the photosensitive element 3a of the CCD image sensor 3 can be almost ignored.

As explained above, the immune reaction component detection device 1 of this embodiment
In this method, a reaction liquid is placed on the top surface of the CCD image sensor 3, and the number of pixels whose light is blocked by aggregates of the reaction liquid is counted.
We perform qualitative and quantitative detection of antibody reactions. Therefore, unlike conventional systems, complicated image processing and optical system mechanisms are not required, resulting in a very simple configuration, and the immune reaction components can be collected by simply placing the reaction liquid on the top surface of the CCD linear sensor 3. can be detected, making the operation extremely easy. Moreover, since the number of aggregates uniformly distributed on a two-dimensional plane is proportional to the number of aggregates dispersed on a one-dimensional plane, a one-dimensional CCD image sensor 3 can be used, and - a simple layer configuration can be used. It becomes.

In addition, in the detection device 1 of this embodiment, the level adjustment knob 1
1 allows the setting of the threshold value VO of the comparator 9 to be changed, so it is possible to cope with brightness that changes depending on the location to be measured. That is, depending on the brightness of the measurement location, the threshold value vO can be set in advance to the best value using a standard reagent, and the immune reaction component can be detected based on the data obtained from the standard material. .

Furthermore, in this example, the CRP standard solution and the CRP sensitized latex solution were stirred using a stirring rod, etc., but the aggregation reaction can be accelerated by applying a high voltage pulse as shown below to the mixed solution. be able to. That is, as shown in FIG. 6, on both ends of the cover glass G on which the mixed solution K was placed,
The electrodes 25 and 27 are installed so as to be immersed in the mixed liquid K, and the high voltage pulse generation circuit 29 generates a voltage between the electrodes 25 and 27.
Apply a high voltage pulse (8K Hz) of 00V.

When a high voltage pulse is applied to the mixed liquid K, the particles in the mixed liquid are dielectrically polarized by the electric field, and collisions between the particles are repeated to promote the aggregation reaction. Moreover, if the application of the high voltage pulse is stopped, only unreacted particles are dispersed.

Although the embodiments of the present invention have been described above, the present invention is not limited to these embodiments.
It goes without saying that CD image sensors 3 having different numbers of pixels and different pixel sizes may be used, and that the present invention can be implemented in various forms without departing from the gist of the present invention.

Furthermore, the object to be tested is not limited to antigens, but may also be antibodies, and a variety of reagents can be used, including not only sensitized latex but also sensitized hemocyte-sensitized gelatin particles.

Effects of the Invention As detailed above, according to the immune reaction component detection device of the present invention, a one-dimensional linear image sensor is used, and the light receiving part is used as a placement part for the reaction liquid, so that a complicated optical system mechanism is required. The present invention has an extremely excellent effect in that it is possible to detect immune reaction components, that is, to perform qualitative or quantitative detection of antigen-antibody reactions with a simple configuration without the need for the following.

[Brief explanation of the drawing]

Figure 1 is a schematic perspective view of the immune reaction component detection device, Figure 2 is a schematic diagram of its configuration, Figure 3 (a) is a plan view of the CCD image sensor, and Figure 3 (b) is its x-X sectional view. , FIG. 4 is a graph showing the output waveform, FIG. 5 is a graph showing the measurement results, and FIG. 6 is an explanatory diagram showing the method of promoting the agglutination reaction. 1... Immune reaction component detection device 3... CCD image sensor 3a... Photosensitive element 3b... Glass plate (light receiving section)
9... Comparator 3... Counter G... Cover glass L... Reaction liquid

Claims (1)

[Scope of Claims] 1. Antigen or antibody in the specimen is determined by measuring the degree of aggregation of the sensitized carrier particles in a reaction solution of the specimen and sensitized carrier particles in which carrier particles are sensitized with antibodies or antigens. In an immunoreactive component detection device that quantitatively or qualitatively detects the presence of An immune reaction component detection device comprising: calculation means for calculating the degree of aggregation of the sensitized carrier particles based on the output value of the sensor.