JPH05297001A - Method and device for automatic immunity measurement using magnetic particle - Google Patents

Abstract

PURPOSE:To obtain a method for measuring immunity for judging flow-out stage of a magnetic particle at the bottom of a container by loading a magnetic particle and a specimen into the container and then allowing them to settle forcibly by magnetic force after agitation and further inclining the container and then an automatic immunity measuring device for performing the method automatically. CONSTITUTION:The title item is provided with a container 26 with a recess on the bottom, a dispensing means for loading a specimen and an antigen or a magnetic particle where an antibody is connected into the container 26, an agitation means for blending the specimen within the container and the magnetic particle, and a settling means for forcibly settling the above magnetic particle by a magnetic body which is laid out at the lower part of the container 26. Further, the title device 1 using the magnetic particle is provided with a container retention means or retaining the container 26 horizontally and slantly and a detection means for detecting the inside of the container 26 after inclination.

Description

Detailed Description of the Invention

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to an automatic immunoassay method and apparatus using magnetic particles.

[0002]

2. Description of the Related Art In JP-A-3-144367 and JP-A-3-191864, magnetic particles having an antigen or an antibody bound thereto and a specimen are put in a container and stirred, and then magnetically forced to settle. There is disclosed an immunoassay method in which the container is tilted and the flowing state of the magnetic particles at the bottom of the container is determined.

In these publications, a device for tilting the container is not specifically shown, and a container in which magnetic particles are forcibly settled is tilted by hand, and the flowing state is visually observed. The flow-out state is judged by comparing with a predetermined flow-out state reference chart.

[0004]

In the above-mentioned conventional immunoassay device, since each step is performed manually, it is difficult to measure a large amount of sample in a short time with high accuracy. The present invention has been made in view of such problems of the conventional immunoassay device, in which magnetic particles and a specimen are put in a container, and the mixture is forcibly settled by magnetic force after stirring,
Another object of the present invention is to provide an immunoassay method for inclining the container to determine the flow-out state of magnetic particles at the bottom of the container, and an automatic immunoassay device for automatically performing this method.

[0005]

The method of the present invention comprises a dispensing step in which a specimen and magnetic particles bound with an antigen or an antibody are placed in a container having a recess at the bottom, and stirring for mixing the specimen and magnetic particles in the container. A step, a settling step of forcibly settling the magnetic particles by a magnetic material, a tilting step of tilting and holding the container, and a detection step for detecting the inside of the container after tilting, This is an automatic immunoassay method using magnetic particles, which is characterized by performing these continuously.

Further, the apparatus of the present invention comprises a container having a recess at the bottom and a dispensing means for putting a magnetic particle (hereinafter referred to as a sensitized magnetic particle) bound with a specimen and an antigen or antibody into the container. A stirring means for mixing the sample in the container and the magnetic particles, a settling means for forcibly settling the magnetic particles by a magnetic substance arranged below the container, and a horizontal and inclined container An automatic immunoassay device using magnetic particles, comprising: a container holding means for holding the container and a detecting means for detecting the inside of the container after tilting.

[0007]

EXAMPLE An automatic immunoassay device using magnetic particles according to an example of the present invention will be described below. As shown in FIG. 1, the automatic immunoassay device 1 includes a container for injecting magnetic particles or a specimen, which has a conical recess having a cone angle of about 120 ° on the bottom from the left on the base 2. For example, a microplate stocker 6 made of plastic, which accommodates a plurality of microplates 4 shown in FIG. 8
A cleaning tank 12 for cleaning the dispensing nozzle 10, a water absorbing paper 14 for absorbing the cleaning liquid adhering to the cleaned dispensing nozzle 10, and a sample plate 1 containing a plurality of samples.
6 and microplate 4 containing the specimen and sensitized magnetic particles
It has a stirring device 17 that vibrates and stirs, a detection unit 18 that tilts the microplate 4 to detect the flowing state of the magnetic particles, and a recovery stocker 20 that recovers the used microplate. Micro plate stocker 6
The structure of is the same as the tray supply device disclosed in, for example, Japanese Patent Publication No. Heisei 2-39478.

The microplate 4 shown in FIG. 3 is provided with depressions in 12 columns and 8 rows, and the concentration of an object to be measured such as hemoglobin decreases in each depression from the left column to the right column. In the first row, it is determined that the sensitizing magnetic particles have not flowed out in the first to fifth columns, and the sensitizing magnetic particles have flowed out in the sixth to eleventh columns.

An operation display panel 22 is provided on the front surface of the base unit 2.
Is provided. A dispensing nozzle operating unit 24 for operating the dispensing nozzle 10 is provided above the cleaning tank 12, the water absorbing paper 14, and the sample plate 16. A container 26 containing a cleaning liquid is disposed in a higher portion behind the recovery stocker 20. As shown in FIG. 2, in the detection unit 18, two mirrors 30 and 32 are arranged above and face each other with an inclination of 45 °. Below the mirror 30, there is a ring light 34 and a CCD / TV camera 36 facing upward.
Is arranged above the ring light 34, and the opal plate 38 for diffusing the beam emitted from the ring light 34.
Are arranged. Instead of the CCD / TV camera 36,
A transmissive photosensor or line sensor can also be used.

Below the mirror 32, there is an inclined plate 39 for receiving the horizontal microplate 4 and holding it at a position shown by an imaginary line inclined by a predetermined angle, for example, 60 °. A solenoid 40 for fixing the microplate 4 is provided on the inclined plate 39, and a permanent magnet 44 for forcibly settling the specimen and magnetic particles dispensed on the microplate 4 is arranged below the solenoid 40. The permanent magnet 44 is
The rack 4 is supported from below by the rack 4
6 is a pinion 6 mounted on the drive shaft 50 of the motor 48
Moves up and down by 0.

Next, the measurement operation of the automatic immunity measuring device 1 having the above-mentioned configuration will be described. These operations are automatically controlled by a program stored in an automatic control device (not shown) such as a microprocessor arranged inside the base unit 2. The microplates 4 accommodated in the microplate stocker 6 are sequentially supplied to the positions shown in FIG. 1 by a belt conveyor (not shown) arranged inside the base 2.

The nozzle operating unit 24 cleans the dispensing nozzle 10 with the cleaning tank 12 and the water-absorbent paper 14 while the reagent tank 8 is being washed.
Of the reagent and the sample on the sample plate 16
Dispense into each reaction recess of. Then, the microplate 4
Is sent to a stirrer 17 by a belt conveyor (not shown) arranged inside the base unit 2, where it is vibrated to mix and stir the specimen in the microplate 4 and the sensitized magnetic particles. It

The microplate 4 is further sent to the detection unit 18 by a belt conveyor (not shown) arranged inside the base unit 2. In the detection unit 18, the permanent magnet 44 first approaches the microplate 4 to force the reacted sensitized magnetic particles to settle in a short time, and then the permanent magnet 44 descends and returns to the initial position. Next, the solenoid 40 is actuated and the tilt plate 39 is tilted by about 60 °, and the microplate 4 is tilted similarly. At this time, the inclination of the conical bottom surface of the microplate 4 is about 30 ° with respect to the horizontal. This tilted state of the microplate 4 is maintained for about 2 minutes, then returned to the original horizontal state, and the flowing state of the specimen and magnetic particles is imaged by the CCD / TV camera 36. An image pickup signal of the CCD / TV camera 36 is image-processed by a microcomputer (not shown) housed in the base unit 2, and a positive sample of
Negative is determined. The result of the determination is the operation display panel 22.
Will be displayed and printed out if necessary.

The determination of positive / negative of the sample will be described with reference to FIG. 4 (A) showing a positive sample flow-out state and FIG. 4 (B) showing a negative sample flow-out state.
The above-mentioned dent is formed along the dotted line aa 'shown in FIG. (A) and the dotted line bb' shown in FIG.
The TV camera 36 sweeps and images. From the image signal, obtain the absolute brightness on the sweep line, measure the length of the particles that flowed out, calculate the rate of change in brightness in the one sweep, calculate the length of the particles that flowed out, of the sensitized magnetic particles Determine the positive / negative by measuring the flow-out area. As a simple determination method, the length of the flow-out portion of the sensitized magnetic particles is measured and compared with a predetermined reference length.

The microplate 4 for which the determination has been completed is a belt conveyor (not shown) arranged inside the base portion 2.
Is sent to the collection stocker 20 and collected. As another embodiment of the present invention, the detection unit is configured to measure the flow-out state of the sensitized magnetic particles while the microplate 4 is tilted, and the CCD unit integrated with the ring light 34 is used.
The TV camera 36 is arranged so as to face the microplate 4 inclined at 60 °.

[0016]

According to the present invention, the sensitized magnetic particles and the specimen are put in a container, and after stirring, the magnetic force is forcibly settled, and the container is tilted to determine the flowing state of the reacted magnetic particles at the bottom of the container. This has the advantage that the immunoassay method can be performed automatically and with high accuracy.

[Brief description of drawings]

FIG. 1 is a perspective view of an automatic immunoassay device of the present invention.

FIG. 2 is a cross-sectional view taken along the line AA of FIG.

FIG. 3 is a plan view of a microplate, showing a state in which magnetic particles have flowed out in a recess in a predetermined measurement state.

FIG. 4 is an explanatory diagram showing a method of determining positive / negative of a sample from a state in which magnetic particles flow out.

[Explanation of symbols]

 1 Automatic Immunoassay Device 2 Base 4 Microplate 6 Microplate Stocker 8 Reagent Tank 10 Dispensing Nozzle 12 Washing Tank 14 Water Absorbing Paper 16 Specimen Plate 18 Detecting Section 20 Recovery Stocker 20 22 Operation Display Panel 24 Dispensing Nozzle Working Section 26 Container 30 Mirror 32 Mirror 34 Ring Light 36 CCD / TV Camera 40 Solenoid 44 Permanent Magnet 48 Motor

Claims (5)

[Claims] 1. A dispensing step in which a specimen and magnetic particles bound with an antigen or an antibody are placed in a container having a recessed portion on the bottom, a stirring step for mixing the specimen and magnetic particles in the container, and a magnetic material It has a settling step of forcibly settling the magnetic particles, a tilting step of tilting and holding the container, and a detecting step for detecting the inside of the container after tilting, and these are continuously performed. An automatic immunoassay method using magnetic particles. 2. A container having a recess at the bottom, a dispensing means for putting a sample and magnetic particles bound with an antigen or an antibody into the container, and a container for mixing the sample and magnetic particles in the container. Stirring means, settling means for forcibly settling the magnetic particles by a magnetic material disposed below the container, container holding means for holding the container horizontally and tilted, and after tilting An automatic immunoassay device using magnetic particles, comprising: a detection unit for detecting the inside of the container. 3. The magnetic particles according to claim 2, wherein the detecting means includes an image pickup device for picking up an image of the inside of the container, and an image processing device for image-processing the output of the image pickup device. Automatic immunoassay device. 4. The automatic immunoassay device using magnetic particles according to claim 1, wherein the detecting means detects the inside of the container after tilting the container while tilting the container. 5. The automatic immunoassay device using magnetic particles according to claim 1, wherein the detecting means detects the inside of the container by making the container horizontal after tilting.