JPH0614048B2 - Enzyme immunological automatic analyzer - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to an enzyme immunoassay automatic analyzer.

In recent years, it has become possible to analyze a very small amount of biological components with the progress of medical treatment, which is useful for early diagnosis of various diseases. For example, various types of incurable diseases such as malignant tumors represented by α-fetoprotein, carcinoembryonic antigen, etc., abnormal secretion of hormones represented by insulin, thyroxine, etc., immune diseases represented by immunoglobulin, etc. Not only early diagnosis of diseases, but also monitoring after treatment of those diseases,
Alternatively, recently, low molecular haptens (incomplete antigens) such as drugs can also be measured, which is useful for preparing drug administration plans.

Many of these biological components are analyzed by an immunochemical method utilizing an antigen-antibody reaction, and various methods have been conventionally proposed as an analyzing method utilizing such an immunochemical reaction. For example, there is a method of analyzing a desired biological component by detecting the presence or absence of an aggregate or the like of the antigen-antibody complex resulting from the antigen-antibody reaction by an agglutination method, a sedimentation method, a turbidimetric method, or the like. However, since these analysis methods require a large amount of antigen-antibody complex and are inferior in sensitivity, they are exclusively used for qualitative analysis or semi-quantitative analysis. Further, in order to make up for the drawbacks of such an analysis method, an antibody or an antigen is bound to fine particles such as carbon particles or a synthetic resin to cause an antigen-antibody reaction with a test substance, and then the agglutination method or turbidimetric method is used. A method of analyzing a test substance, or a labeled antibody or antigen labeled with a highly sensitive marker such as a radioisotope, a fluorescent substance, a luminescent substance or an enzyme on the antibody or antigen, is used to detect the antigen-antibody complex with high sensitivity. A method of detecting and analyzing a test substance has also been proposed. However, since the former method using fine particles is inferior in sensitivity to the latter method using a marker, recently, the latter analysis method using a marker having high detection sensitivity has become mainstream.

As an analysis method using such a marker, a radioimmunoassay method using a radioisotope as a marker, a fluorescence immunoassay method using a fluorescent substance, an enzyme immunoassay method using an enzyme, etc. are known. However, the enzyme immunoassay does not require any special equipment or measurement technique and can be easily performed using a colorimeter that is widely used, and thus has recently attracted particular attention. This enzyme-linked immunosorbent assay is a homogeneous (Homoge) method in which the amount of change in the activity of the labeled enzyme is directly determined depending on the presence or absence of an immunochemical reaction to quantify the test substance.
neous) enzyme immunoassay, and the enzyme-labeled antibody that has reacted with the antigen or the antibody or the enzyme-labeled antigen and the unreacted one are subjected to BF separation by a washing operation, and the activity amount of the labeled enzyme after the BF separation is determined. Heterogeneous enzyme immunoassay for quantifying test substances. However, the former homogeneous enzyme immunoassay method can be carried out by a simple operation, but has a drawback that it can analyze only low molecular haptens such as drugs, and cannot analyze biological components that are macromolecules. On the other hand, the latter heterogeneous enzyme immunoassay method requires a washing operation for performing B / F separation, but can properly analyze whether the test substance is a low molecule or a high molecule, It is being generalized because its analysis target is extremely wide.

As such a heterogeneous enzyme immunoassay method, a competitive method, a sandwich method and the like are known. Competitive law is the first
As shown in the figure, for example, an insoluble bead 1 made of synthetic resin such as plastic or glass is immobilized in advance with an antibody or an antigen that causes an antigen-antibody reaction with the test substance in the sample.
The beads 1 and the sample and the labeling reagent 3 which is the same substance as the test substance 2 are enzyme-labeled in a reaction container to cause an antigen-antibody reaction, and then washing is performed to compete with the beads 1 by the antigen-antibody reaction. A color-developing reagent that reacts with the labeling enzyme in the labeling reagent 3 in a reaction container containing the beads 1 after BF separation of the analyte 2 and the labeling reagent 3 that are bound to each other and those that are not bound After adding and reacting, the reaction solution is subjected to colorimetric measurement to obtain the enzyme activity of the labeling enzyme to quantify the test substance 2. As shown in FIG. 2, the sandwich method uses insoluble beads 5 on which an antibody or an antigen that causes an antigen-antibody reaction with a test substance in a sample is immobilized in advance, as shown in FIG. And the sample are housed in a reaction container to cause an antigen-antibody reaction to bind the test substance 6 in the sample to the beads 5, and then washing is performed to perform B / F separation, and then the beads 5 are housed. The reaction container containing the test substance 6 and the labeling reagent 7 in which a substance that causes an antigen-antibody reaction is labeled with an enzyme are housed to cause an antigen-antibody reaction, and then washed again to perform BF.
After separation, a coloring reagent that reacts with the labeling enzyme in the labeling reagent 7 is added and reacted, and then the reaction solution is colorimetrically measured to obtain the enzyme activity of the labeling enzyme to quantify the test substance 6. is there.

The above-mentioned heterogeneous enzyme immunoassay is performed by a conventional method, but recently, its automation is being promoted. For this automation, a method of repeatedly using a reaction container for carrying out an antigen-antibody reaction and a method of disposing it are considered. In FIGS. 1 and 2, the antibody or antigen is immobilized on beads. Therefore, when adopting the former method, it is necessary to put beads into the reaction container, take out the beads remaining in the reaction container after the analysis, and wash the reaction container for reuse. The operation is complicated and there is a risk of contamination due to reuse of the reaction vessel. Further, when the latter method is adopted, the beads and the reaction container are discarded after the analysis is completed, so that the running cost per sample becomes high.

Furthermore, in the above-mentioned heterogeneous enzyme immunoassay, the B.F. value is once in the competitive method and the one-step sandwich method and twice in the ordinary two-step sandwich method during the analysis of one test substance. Separation is required. In addition, when the reaction container is supplied to the reaction line for analysis, and after the completion of the analysis, it is discarded from the reaction line and disposed of, especially when using the one in which a predetermined antibody or antigen is immobilized on the inner wall of the reaction container. In addition, the reaction container supplied to the reaction line can be washed prior to the analysis, and in this case, one additional washing step is added, and in the above-mentioned enzyme immunoassay method, one test substance Analysis requires at least two washing steps including BF separation. In this case, it is conceivable to dispose a dedicated cleaning device for each cleaning process, but this causes a problem that the device becomes large, complicated, expensive, and complicated in control.

An object of the present invention is to solve the above-mentioned various problems, to make the apparatus small and simple, to be inexpensive, and to easily control, and to continuously analyze the test substance in the sample on the reaction line. The present invention provides an immunological automatic analyzer capable of

In order to achieve this object, in the present invention, in an enzyme immunological automatic analyzer, an apparatus for holding a plurality of reaction vessels in which a predetermined antibody or antigen is immobilized on at least a part of the inner wall, and the reaction vessel are connected in series. Device, a device for confirming the presence or absence of a reaction container, a device for dispensing a reagent into the reaction container supplied to the reaction part, a device for dispensing a sample into the reaction container supplied to the reaction part, A cleaning device for removing reaction substances, a device for measuring the reaction liquid still contained in the reaction container, a device for discharging the reaction container after the measurement from the reaction part, and a reaction container for discarding the reaction container And a storage device.

Hereinafter, the present invention will be described in detail with reference to the drawings.

FIG. 3 is a diagram showing the configuration of an example of the enzyme immunological automatic analyzer of the present invention, which employs the sandwich method described in FIG. A turntable 11 as a processing table has 37 reaction vessels (hereinafter referred to as cuvettes) 12 each having a predetermined antibody or antigen immobilized on a small portion of the inner wall at the periphery thereof and attached / detached at equal intervals on the same circumference. A cuvette holder 13 which can be freely held is provided, and the cuvette 12 held by the holder 13 is intermittently rotated at a predetermined pitch (for example, 15 seconds) in a direction indicated by an arrow in a horizontal plane. The cuvette holder 13 by the intermittent rotation of the turntable 11
The stop positions of the cuvette holding part are shown by reference symbols S _{1 to} S ₃₇ . In this example, at the stop position S ₁ , the cuvette supply device 15 continuously supplies the cuvettes 12 to the cuvette holder 13 at a predetermined pitch and holds them. Next stop position S
_{In 2} , the presence or absence of the cuvette 12 is detected by a detection means (not shown), and when it is detected that the cuvette 12 is held, the washing device 16 is used to exchange ion-exchanged water, an immunoassay buffer, physiological saline, etc. The cleaning liquid in (1) is injected and discharged to separate B and F and the cuvette 12 is cleaned.

Also, at the stop position S ₄ , the first reagent 18 is used by the first reagent dispensing device 17, the third reagent 20 is used by the third reagent dispensing device 19 at the stop position S ₅ , and the second reagent is used at the stop position S ₆ . the second reagent 22 by the reagent dispensing apparatus 21, first in samples samples from the cup 25 and stop position S _29, in the predetermined sample suction position of the sampler 24 by the sample dispensing unit 23 at the stop position S ₇ 4 Reagent dispenser 26
Thus, the fourth reagent 27 is continuously dispensed at a predetermined pitch. Here, as the first reagent 18, a buffer solution, as the second reagent 22, an enzyme labeling reagent corresponding to the test substance in the sample, and as the third reagent 20, a coloring reagent that develops a color by reacting with a labeling enzyme, As the fourth reagent 27, reaction stopping reagents that stop the reaction between the labeling enzyme and the coloring reagent are used. Further, although the sampler 24 can be of any type, it is originally 10 sample cups 25 each.
Holding a number of racks 24a arranged side by side, the racks on the left side of the column are moved sequentially downward in FIG. 3 to the sample dispensing position, and the racks on the right side for holding the sample cups after dispensing are completed. Move the rack upwards. The rack at the sample dispensing position is intermittently moved in the direction of arrow S in synchronization with the rotation of the turntable 11. When all the samples held in this rack have been dispensed, this rack is sent to the bottom of the right rack row and the bottom rack in the left row is then sent to the sample dispense position. . In this way, successive samples can be continuously sent to the sample dispensing position at a predetermined pitch.

Further, at the stop position S ₃₂ , the final test solution into which the fourth reagent 27 has been dispensed is colorimetrically measured by the colorimetric device 28, and the cuvette 12 for which the colorimetric measurement has been completed is stopped at the stop position S ₃₅ . It is removed from the cuvette holder 13 by 29. In this example, the colorimetric device 28 directly measures the test liquid through the cuvette 12, and the cuvette discharge device 29
In (1), the cuvettes 12 dropped from the cuvette holder 13 are separated from the test liquid and housed in respective discarding containers, and then the cuvettes are discarded.

Next, the operation of the enzyme immunoassay automatic analyzer shown in FIG. 3 will be described with reference to FIGS. 4 and 5.

In this example, the sandwich method is used for analysis.
Each sample is analyzed by spinning and performing one wash prior to analysis and two BF separations during analysis. Therefore, sample dispensing, first, second, third, and fourth reagent dispensing, cuvette 12 supply, discharge, colorimetric measurement, etc. are performed once while the turntable 11 moves 3 pitches. It is supposed to do. However, since the cleaning is performed three times for each cuvette 12 as described above, the cleaning is performed for each moving pitch of the turntable 11. Therefore, the presence or absence of the cuvette 12 at the stop position S ₂ is detected to detect the cleaning device. 16
Cuvette detection means for actuating the.
Also, in order to operate in this way, the cuvette holder
The number of cuvettes 12 that can be loaded in 13 is 1, 2, 3,
..., it is necessary to use 3n + 1 or 3n + 2. In this example, n = 12 is set so that 37 cuvettes 12 can be loaded in the cuvette holder 13.

In the first rotation of the turntable 11, first, at the stop position S ₁ , as shown in FIG.
The cuvette 12 in which a predetermined antibody or antigen is immobilized on at least a part of the inner wall is supplied and held in the cuvette holder 13, and the cuvette 12 is detected by the detection means (not shown) at the next stop position S ₂ and the washing device 16 To wash prior to analysis. At the stop position S ₄ , where the cuvette 12 is supplied at the stop position S ₁ and sent for 3 pitches, at the stop position S ₄ , the first reagent dispensing device 17 dispenses a predetermined amount of the first reagent 18 made of a buffer solution, and then further 3 pitches are sent. At the stopped position S ₇ , the sample dispensing device 23 dispenses a predetermined amount of the sample, and the first antigen-antibody reaction is started. In FIG. 5, the operation dimming performed on the cuvette is indicated by a diagonal line descending to the left.

Next, the turntable 11 enters the stop position S in the second rotation.
_{In FIG. 2} , after the washing by the washing device 16, that is, the first B / F separation is performed, the stop position S ₆ is reached, and the cuvette 12 is filled with the enzyme-labeled reagent by the second reagent dispensing device 21. A certain second reagent 22 is dispensed in a predetermined amount, and the second antigen-antibody reaction is started.

Further, the turntable 11 reaches the stop position S ₅ after cleaning by the cleaning device 16 at the stop position S ₂ in the third rotation, that is, the second BF separation.
Here, the third reagent dispensing device 19 dispenses a predetermined amount of the third reagent 20, which is a coloring reagent, into the cuvette 12 to start the coloring reaction. Next, the cuvette 12 is stopped at the stop position S _29.
The fourth reagent dispensing device 26 dispenses a predetermined amount of the fourth reagent 27, which is a reaction stop solution, to start the color reaction, and then the cuvette is stopped at the stop position S ₃₂ after three pitches. The test solution contained in 12 is directly measured by the colorimetric device 28 through the cuvette 12. This cuvette 12 has 3 pitches of the turntable 11 that has been fed 3 pitches.
At the final stop position S ₃₅ of the second time, the cuvette discharge device
The cuvette is discharged from the cuvette holder 13 by 29.
After separating 12 and the test solution into their respective disposal containers, discard them. In this way, the cuvette holding portion from which the cuvette 12 has been discharged at the stop position S ₃₅ is further fed by 3 pitches and reaches the stop position S ₁ , where the cuvette supply device 15 receives the supply of a new cuvette.
The operation dimming for this new cuvette is shown by the diagonal line to the lower right in FIG.

As described above, the analysis operation for one cuvette is completed by the turntable 11 rotating approximately three times. In this example, sample dispensing, first, second, third and fourth reagent dispensing are performed. For supplying, discharging, and colorimetric measurement of cuvettes, the turntable 11 moves once by moving 3 pitches, and 37 (3 × 12 + 1) cuvettes 12 can be mounted on the turntable 11 at equal intervals, so stop, for example. The cuvette positioned when the sample dispensing apparatus 23 operates at the position S ₇ is displaced by one for each rotation of the turntable 11. Since such a situation can be said for all the operations that are performed once for three pitches, sample dispensing can be continuously performed once for every three pitches. Therefore, the ID control of the sample, the processing of the analysis result, and the like can be performed at a constant cycle, and various controls are facilitated. Furthermore, the reaction line is made endless, and the cuvette 12 is circulated and conveyed to one cleaning device 16 provided in the reaction line to repeatedly perform cleaning including BF separation, so the overall structure of the device is small and simple. And can be made inexpensive. In addition, since the antibody or antigen is immobilized on a small part of the inner wall of the cuvette 12, it is necessary to use beads on which the antibody or antigen is immobilized as shown in FIGS. 1 and 2 separately from the reaction vessel. Compared with this, running cost can be reduced.

FIG. 6 is a diagram showing the construction of another example of the enzyme immunoassay automatic analyzer of the present invention, and the same components as those shown in FIG. 3 are designated by the same reference numerals. In this example, the competitive method described with reference to FIG. 1 is adopted, and in this case, a sample and an enzyme labeling reagent were added to a cuvette to which a predetermined antigen or antibody was bound to carry out an antigen-antibody reaction. Since the subsequent BF separation is performed and then the colorimetric measurement is performed after adding the enzyme coloring reagent, the cuvette is washed twice before the analysis. Therefore, in the example shown in FIG. 6, 2n + 1 cuvettes 12 can be loaded in the cuvette holder 13 of the turntable 11, and the sample dispensing, reagent dispensing, cuvette supply and discharge, colorimetric measurement, etc. are performed by the turntable 11. Is operated once every two pitches are moved, the sample can be dispensed continuously, that is, every two pitches.

In FIG. 6, the cuvette 12 in which a predetermined antibody or antigen is immobilized is supplied to a small part of the inner wall by the cuvette supplying device 15 at the stop position S ₁ , and the presence or absence of the cuvette 12 is detected at the stop position S ₃ . Then, the cuvette 12 supplied by the cleaning device 16 is cleaned and BF separation is performed. Further, at the stop position S ₅ , the first reagent dispensing device 21
The first reagent 22, which is an enzyme labeling reagent, is dispensed by a predetermined amount, and the sample is dispensed by the sample dispensing device 23 by a predetermined amount. Also in this example, the sample is sequentially supplied to the sample dispensing position from the sampler 24 having a plurality of racks 24a holding a plurality of sample cups 25. Further, at the stop position S ₆ , the second reagent dispensing device 19 dispenses a predetermined amount of the second reagent 20 which is a coloring reagent, and at the stop position S ₃₂ , the third reagent dispensing device 26 serves as a reaction stop solution. Third
A predetermined amount of reagent 27 is dispensed. Furthermore, at the stop position S ₃₄ , the colorimetric device 28 directly measures the final test liquid through the cuvette 12, and at the stop position S ₃₆ , the cuvette discharge device 29 finishes the colorimetric measurement.
Remove the 12 from the cuvette holder 13
Then, the test solution is separated from the test solution, housed in each disposal container, and then discarded.

FIG. 7 shows the operation timing of the automatic analyzer shown in FIG. A certain cuvette supplied at the stop position S ₁
Looking at the analysis operation for 12, first, the washing device 16 performs washing prior to the analysis at the stop position S ₃ sent by 2 pitches, and then the first reagent dispensing device 21 at the stop position S ₅ sent by 2 pitches. The sample dispensing device 23 dispenses the first reagent 22 and the sample, which are enzyme labeling reagents, respectively, and the antigen-antibody reaction is performed. Next, in the second rotation, the cuvette is detected by the detection means (not shown) at the stop position S ₃ , and the cleaning device 16 performs cleaning, that is, BF separation, and then the second reagent dispensing at the stop position S ₆ . The device 19 dispenses the second reagent 20, which is a coloring reagent, to carry out the coloring reaction. This color reaction is stopped at the stop position S _{32 by} the third reagent dispensing device 26 dispensing the third reagent 27, which is the reaction stop solution, and the cuvette 12 reaches the stop position S ₃₄ and the test solution is obtained. Is colorimetrically measured by the colorimetric device 28 through the cuvette 12. After that, at the stop position S ₃₆ which is fed by two pitches, the cuvette 12 is discharged from the cuvette holder 13 by the cuvette discharging device 29. In FIG. 7, the operation timing performed on the cuvette is shown by a downward-sloping diagonal line, and the operation timing performed on the cuvette newly supplied to the cuvette holding portion after discharging the cuvette is lowered to the right. Is indicated by the diagonal line. In this way, by rotating the turntable 11 about two times for each cuvette, a predetermined analysis of each sample can be performed. Also, in this example, the turntable 11
Since 37 cuvettes 12 can be loaded in the cuvette holder 13 of the above, and sample dispensing, reagent dispensing, cuvette supply, discharge, and colorimetric measurement are performed once every 2 pitches, 2 consecutive samples are collected. It is possible to dispense continuously in a pitch cycle. However, the cleaning device 16 must operate at each pitch after the turntable 11 makes one rotation.

Specific configurations of the respective parts of the embodiment shown in FIGS. 3 and 6 will be described below.

8A to 8C show an example of the structure of the cuvette 12. FIG. 8A is a perspective view and FIG. 8B is I-I of FIG. 8A.
FIG. 8C is a sectional view taken along line II-II of FIG.
In this example, the cuvette 12 is a molded product of transparent synthetic resin, has a flat box shape as a whole, and has an opening 12 at the top.
While a is formed, a predetermined antibody or antigen is immobilized on at least a part of the inner wall. One major surface 12
A T-shaped protrusion 12c is formed integrally with b, and the cuvette 12 is held in the cuvette holder 13 by utilizing the elastic restoring force of the protrusion 12c as described later. Cuvette
The side walls 12d and 12e of 12 are arranged perpendicular to the side optical axis, and the lower side of these side walls are slightly recessed to form an entrance window 12f and an exit window 12g. By thus forming the entrance window and the exit window by denting from the side wall, it is possible to prevent this portion from being soiled or scratched by hand, and to reduce the incidence of stray light from the surroundings. Also, FIG. 8C
As clearly shown in FIG. 2, the lower part of the cuvette 12 has a semi-cylindrical shape. Therefore, if an antibody or an antigen is immobilized on this part, effective photometry can be performed with a small amount of test solution. The antibody or antigen can be immobilized on the cuvette 12 made of such synthetic resin by a physical adsorption method or a chemical bonding method. The cuvette 12 is not limited to synthetic resin and may be made of glass, and in this case, the antibody or antigen can be immobilized by a chemical bonding method such as a covalent bonding method. Further, when the antibody or the antigen, or the antigen or the protein such as the antibody or the labeling enzyme which binds to the antibody or the antigen binds to the measurement light, the immobilization site of the antibody or the antigen has a portion of the entrance window 12f and the exit window 12g. It should be excluded.

FIG. 9 is a colorimetric device using the cuvette 12 shown in FIG.
28 shows an example of the configuration of 28. In this example, the light from the light source lamp 28a is condensed by the lens 28b, is made incident on the entrance window 12f of the cuvette 12 via the diaphragm 28c, and the light emitted from the exit window 12g is made by the diaphragm 28d and the optical filter.
The light is incident on the detector 28f via 28e. The cuvette 12 has a notch 13 formed on the periphery of the cuvette holder 13.
It is elastically fitted and held in a.

FIG. 10 is a perspective view showing the structure of an example of a magazine that stores a large number of the cuvettes 12 described above. The cuvette 12 is not stored in the magazine 30 by the user, and the user can obtain the magazine in which the cuvette 12 is stored.
12 damage and fingerprints can be more reliably prevented. The magazine 30 can be a synthetic resin molded product or a metal product, and in this example, ten magazines are arranged in the lateral direction indicated by the arrow A, and ten magazines are formed in the vertical direction indicated by the arrow B, which is a total of ten magazines. An array of 100 cuvettes 12 is stored. An outlet 31b having a width substantially equal to the thickness of the cuvette 12 is formed at one end of one side wall 31a of the magazine 30. Cuvette 12 is this exit
A narrow front side wall so that it can be discharged from 31b in the correct posture
A protrusion 31 having elasticity in a portion adjacent to the outlet 31b of 31c
form c. The magazine 30 is further formed with three grooves 31f extending from the upper wall 31d to the rear wall 31e. Further, a pressing plate 32 is interposed between the cuvette array and the rear side wall 31e, and the pressing plate 32 is moved in the arrow B direction as described later, so that the cuvette array can be moved in the B direction at the same time. ing. Further, a notch 31g is formed at the left end of the side wall 31a, and when the magazine 30 is loaded in the cuvette feeder, the protrusion of the cuvette feeder is notched 31g.
To prevent reverse insertion.

FIGS. 11 and 12 show an example of the configuration of a cuvette supply device 15 for loading the cuvettes 12 housed in the magazine 30 into the notches (cuvette holding portions) 13a of the cuvette holder 13 one by one. Yes, FIG. 11 is a plan view, and FIG. 12 is a sectional view. The turntable 11 is rotated in a direction indicated by an arrow at a predetermined pitch, and a cuvette holder 13 having a large number of notches 13a formed at equal intervals is provided on the periphery of the turntable 11.

The cuvette supply device 15 includes a substrate 40, and a magazine storage portion 41 is fixed to the bottom surface of the substrate 40 as shown in FIG. A magazine receiver 42 is arranged in the magazine storage unit 41 so as to be vertically movable. The magazine receiver 42 is a pulley.
One end of a wire 44 laid over 43 is fixed, and the other end of the wire is fixed to a weight 46 movably arranged in a cylindrical guide 45. The guide 45 supports the magazine receiver 42 via a linear bearing 47. Therefore magazine receiving 42
Will always be biased upwards. Magazine compartment 41
As shown in FIG. 12, a plurality of magazines 30 accommodating a large number of cuvettes 12 can be loaded therein.

A first opening 40a through which the magazine passes is formed in the substrate 40 above the magazine housing portion 41. On the upper surface of the substrate 40, a pair of L-shaped levers 48 and
49 are provided so as to be rotatable about axes 48a and 49a, respectively. Ring-shaped stoppers 50 and 51 are attached to these levers 48 and 49 by shafts 50a and 51a, respectively, and rollers 52 and 53 for rotating these levers are attached by shafts 52a and 53a as described later. Further, the free ends of these levers 48 and 49 have engaging projections.
Form 48b and 49b. L-shaped columns 54 and 55 are further provided on the side of the first opening 40a of the substrate 40, and stoppers 54a and 55a are attached to the tips of these columns.

The substrate 40 is further formed with a second opening 40b through which the magazine passes. A pair of magazine receiving levers 56 and 57 are provided on the sides of the second opening 40b so as to rotate about shafts 56a and 57a, respectively. Pins 56b and 57b are planted near the free ends of these levers 56 and 57, respectively, and these pins are fitted to the engaging projections 48 of the levers 48 and 49.
engage b and 49b. Pins 56c and 57c are further planted on the levers 56 and 57, and the pins of the push levers 58 and 59 which rotate about axes 58a and 59a extending parallel to the plane of FIG. Engage. A spring is attached to each of these levers 48, 49, 56, 57, 58 and 59, and the levers are biased to the positions shown by the solid lines. The push levers 58 and 59 rotate in a plane perpendicular to the plane shown in FIG. 11 when the levers 56 and 57 are displaced as shown by an imaginary line.

Two guide shafts 60a and 60b extending above the first and second openings 40a and 40b are fixed to the substrate 40 via leg portions 61a and 61b. These pair of guide shafts 60a
And 60b are provided with a first slider 62 slidably via a linear bearing, and a wire is attached to the first slider 62.
One end of 63 is fixed, and this wire is passed over the pulley 64 attached to the leg 61a, the pulley 66 attached to the drive shaft of the motor 65 and the pulley 67 attached to the leg 61b, and the other end is also fixed to the slider 62. To do. Therefore, by reversibly rotating the motor 65, the slider 62 is moved to the guide shafts 60a and 60b.
It is possible to reciprocate in the B direction along. This movement is carried out by the magazine 30 located above the first opening 40a.
For moving the cuvette 21 in the magazine 30 in the B direction while moving the cuvette 21 to the loading position above the second opening 40b. Therefore, as shown in FIG. 12, under the slider 62, it is possible to enter the groove 31f formed in the magazine 30.
Attach book arm 62a.

Further, a pair of guide shafts 68a and 68b extending in a direction orthogonal to the guide shafts 60a and 60b along the side edge of the second opening 40b are attached to the substrate 40 via leg portions 69a and 69b. A second slider 70 is slidably provided on the guide shaft, one end of a wire 71 is fixed to the slider 70, and the wire 72 is provided on a leg portion 69a.
It is passed around a pulley 74 attached to the drive shaft of the motor 73 and a pulley 75 attached to the leg portion 69b, and the other end is fixed to the slider 70. Therefore, the slider 70 can be moved in the direction A along the guide shafts 68a and 68b by rotating the motor 73 forward and backward, whereby the cuvettes 12 from the magazine 30 are inserted into the notches 13a of the cuvette holder 13 one by one. Can be loaded. For this purpose, the slider 70 has a pin 70
A is provided, a pushing claw 70c is fixed to the tip thereof, and a coil spring 70b is inserted into the pin 70a so that the pushing claw 70b can push the side wall of the cuvette 12 at the end.

A coil spring 76 is inserted between the guide shaft 68b and the leg portion 69a, and the guide shafts 68a and 68b are slidable with respect to the leg portion 69b. Therefore, the guide shafts 68a and 68b are shown in FIG. Bias to the left in the plane. Further, as shown in FIG. 12, the slider 70 is slidably inserted into the guide shaft 68a through a linear bearing, but is frictionally engaged with the guide shaft 68b by a coil spring 77 and a ball 78. Therefore, the slider 70 and the guide shaft 68b move together over a certain range. An L-shaped rail receiver 79 is fixed to the other end of the guide shaft 68b, and a guide rail 80 having a recess having a width substantially equal to the thickness of the cuvette 12 is fixed to the rail receiver. The guide rail is guided by guide rollers 81a to 81d and slightly reciprocates in the A direction. A pressing spring 82 for pressing the cuvette at the tip is attached near the tip of the guide rail 80.

Next, the operation of the cuvette supply device 15 will be described. For convenience of explanation, several magazines 30 are provided in the magazine storage section 41.
The top of the magazine in the uppermost position is loaded with the stopper 50 and the stopper 50 attached to the levers 48 and 49 in the solid line position.
It shall be engaged with 51. The magazine 30 is located above the second opening 40b, and the lever is in the solid line position.
It is supported by 56 and 57 so that it does not fall downward. That is, the magazine 30 is in the cuvette loading position, and the cuvettes 12 accommodated therein can be sequentially inserted into the notches 13a of the cuvette 13. In other words, by rotating the motor 73 forward,
71 moves counterclockwise in FIG. 11, and the slider 70 moves in the A direction accordingly. At this time, the guide shaft 68b is also A
The rail receiver 79 and the guide rail 80 also move in the A direction. At this time, the cuvette row (the cuvettes arranged horizontally in the uppermost direction in FIG. 11)
Also moves in the A direction. Next, the nut 68c at the left end of the guide shaft 68b
When the abuts on the leg portion 69a, the guide shaft 68b no longer moves, and only the slider 70 moves in the A direction. As a result, the cuvette row is further pushed out in the direction A, the cuvette at the right end is disengaged from the guide rail 80, and the notch of the cuvette holder 13 is cut out.
It is inserted into 13a. As described above, since the cuvette 12 is formed with the elastic protrusion 12c, it is elastically fitted in the notch 13a. Next, when the motor 73 is rotated in the reverse direction, both the slider 70 and the guide shaft 68b return in the direction opposite to the A direction, and the rail receiver 79 is brought into contact with the leg portion 69b. During this time, the push claw 70c of the slider 70 remains in contact with the cuvette. By repeating the above operation at a predetermined timing, successive cuvettes can be inserted into the notches 13a of the cuvette holder 13. When the insertion of the uppermost cuvette row is completed, the motor 73 is reversed and the slider 70 is returned to the left end position. Next, the motor 65 is rotated forward by a predetermined amount, and the wire 63
Is rotated clockwise to move the slider 62 in the B direction by a predetermined pitch (equal to the thickness of the cuvette). As a result, the cuvette in the magazine 30 is pushed by the push plate 32 and moves upward at the 11th position.

Next, when the motor 65 is normally rotated, the slider 62 moves upward in FIG. 11, and the levers 48, 49, 56, 57, 58 and 59 return to the positions indicated by the solid lines. In this way, a new magazine can be transferred to the cuvette loading position.

The cuvette discharging device 29 described above uses the solenoid 13 to cut the cuvette 12 that is elastically fitted and held in the cutout 13a of the cuvette holder 13, for example.
The cuvette 12 is discharged from a, and the discharged cuvette 12 is dropped while rolling along an inclined surface made of a mesh, so that the cuvette 12 and the test solution are separated and housed in respective disposal containers. it can.

It should be noted that the present invention is not limited to the above-mentioned examples, and many modifications and variations are possible. For example, since the cuvette is disposable, it does not necessarily need to be washed prior to analysis. Further, in the above-described example, the finally obtained test solution is colorimetrically measured by the direct photometric method through the cuvette, but the test solution in the cuvette can be guided to the colorimetric cell for colorimetric measurement. . In this case, since the coloring reaction is stopped by introducing the test solution into the colorimetric cell, the dispensing of the reaction stop solution can be eliminated. Further, although one cleaning device is provided in the above-described embodiment, a plurality of cleaning devices may be provided. For example, in the embodiment shown in FIG. 3, the second cleaning device may be provided at a position substantially opposite in diameter to the cleaning device 16. Even in this case, the effect that the apparatus is simple and small can be obtained as compared with the case where three cleaning apparatuses are provided. Furthermore, in the above-mentioned embodiment, the same measurement item was analyzed for all samples, but a large number of cuvette supply devices and labeling reagent dispensing devices in which antibodies or light sources are immobilized according to the measurement items are provided. It is also possible to carry out multi-item analysis at the same time. In this case, the reaction line may be one line or the number of lines corresponding to the number of measurement items. In addition, various dispensing positions, cuvette supply,
The discharging position, the colorimetric measuring position, etc. are not limited to those in the above-described embodiment, and various changes can be made. Further, although the stirring of the above example is not mentioned at all, an appropriate stirring mechanism can be provided at an appropriate stop position.
Further, although the cuvette has a flat box shape in the above-mentioned example, it may have another shape. Further, in order to stably carry out the reaction in the cuvette supplied to the reaction line, the cuvette may be transported while being immersed in a constant temperature bath in the reaction line.

As described above, according to the present invention, since the reaction container is disposable, it is possible to effectively prevent contamination between samples, and the simple control of supplying and discharging the reaction container every several pitches. Since the reaction vessel is continuously supplied and discharged, sample analysis can be efficiently performed. In addition, during each sample analysis, the reaction vessel is conveyed to the same washing position in the reaction line several times, and B.
Since the washing including the F separation is performed a plurality of times, it is possible to reduce the number of washing devices, and therefore, it is possible to obtain an effect that the analyzer is small and inexpensive, and that the configuration can be simplified.

[Brief description of drawings]

FIG. 1 is a diagram showing a process of enzyme immunoassay by a competitive method using beads, FIG. 2 is a diagram showing a process of enzyme immunoassay by a sandwich method also using beads, and FIG. 3 is an enzyme immunoassay of the present invention. Diagram showing the construction of one embodiment of a biological automatic analyzer, FIG. 4 is a diagram showing its sequential operation, FIG. 5 is a timing chart diagram showing the operation of each part thereof, and FIG. 6 is the present invention. FIG. 7 is a diagram showing the configuration of another embodiment of the enzyme immunological automatic analyzer of the present invention, FIG. 7 is a timing chart diagram for explaining the same operation, and FIGS. 8A to 8C show reaction vessels used in the present invention. FIG. 9 is a diagram showing an example of the configuration, FIG. 9 is a diagram showing an example of the configuration of the colorimetric device, and FIG. 10 is a perspective view showing an example of the configuration of a magazine for accommodating a large number of reaction vessels shown in FIG. Figures 11, 11 and 12 show an example of a reaction container supply device. It is the top view and sectional drawing which show a structure. 11 …… Turntable 12 …… Reaction container (cuvette) 13 …… Cuvette holder 15 …… Cuvette supply device 16 …… Washing device 17,19,21,26 …… Reagent dispensing device 23 …… Sample dispensing device 24 …… Sampler 25 …… Sample cup 28 …… Colorimetric device 29 …… Cuvette discharge device

 ─────────────────────────────────────────────────── ─── Continuation of the front page (56) Reference JP-A-57-74662 (JP, A) JP-A-56-147067 (JP, A) New medical treatment Vol. 8 No6 (1981) P. 87-90

Claims (1)

[Claims] 1. An enzyme-immunoassay automatic analyzer for holding a plurality of reaction vessels each having a predetermined antibody or antigen immobilized on at least a part of its inner wall, and an apparatus for continuously supplying the reaction vessels. , A device for checking the presence or absence of a reaction container, a device for dispensing a reagent into the reaction container supplied to the reaction part, a device for dispensing a sample into the reaction container supplied to the reaction part, and for removing unreacted substances A cleaning device, a device for measuring the reaction solution still contained in the reaction container, a device for discharging the reaction container after the measurement from the reaction part, and a reaction container housing device for discarding the reaction container. An enzyme immunological automatic analyzer characterized by: