JPS59135367A - Immunological automatic analytical method - Google Patents

Abstract

PURPOSE:To continuously perform the distribution of a specimen, by performing the washing of a reaction tube by recirculating and passing the same through the same washing apparatus during the analysis of each specimen by recirculating and passing the reaction tube through the washing apparatus provided to a reaction line constituted in an endless form during the analysis of each specimen while continuously distributing the specimen. CONSTITUTION:In the first rotation of a reaction tue disc 12, one carrier 23 from a carrier throw-in apparatus 22 is thrown in the U-shaped tube 11 positioned at a stop position S1 from the large opening part 11a of said tube 11. At this stop position S1, a predetermined amount of a first reagent 17 comprising a buffer solution is simultaneously distributed in the tube 11 by a first reagent distributing apparatus 16. After the reaction tube 11 is sent over by three pitches, a predetermined amount of a specimen is distributed in the tube 11 at a position S4 by a specimen distributing apparatus 13. By this procedure, antigen-antibody reaction is started. The reaction tube reaches a stop position S25 at the last stage of the first rotation and washing is performed by a washing apparatus 26 while a first time B.F separation is performed. In the next step, the reaction tube disc 12 enters a second rotation and, at a stop position S3, second reaction is started. At the final stop position S25 of the second rotation, second B.F separation is performed by a washing apparatus 25.

Description

[Detailed description of the invention] The present invention relates to an automated immunological analysis method.

In recent years, advances in medical care have made it possible to analyze minute amounts of biological components, which is useful for early diagnosis of various diseases. For example, various diseases considered to be incurable, such as malignant tumors represented by α-fetoprotein and carcinoembryonic antigen, abnormal hormone secretion diseases represented by insulin and thyroxine, and immune diseases represented by immunoglobulin, etc. Not only can diseases be diagnosed early, but they can also be monitored after treatment,
Recently, it has become possible to measure low-molecular haptens (incomplete antigens) such as drugs, which is also useful in creating drug administration plans.

Many of these biological components are analyzed by immunochemical methods that utilize antigen-antibody reactions, and various methods have been proposed in the past as analytical methods that utilize such immunochemical reactions. For example, there are methods of detecting the presence or absence of aggregates of antigen-antibody complexes produced as a result of antigen-antibody reactions by agglutination methods, sedimentation methods, turbidimetry, etc., and analyzing desired biological components. However, these analytical methods require a large amount of antigen-antibody complexes and are inferior in sensitivity, so they are used exclusively for qualitative or semi-quantitative analysis. In order to compensate for the shortcomings of such analysis methods, antibodies or antigens are bound to fine particles such as carbon particles or synthetic resins, and antigen-antibody reactions are performed with the test substance, followed by agglutination or turbidimetry. The method of analyzing the test substance and the use of labeled antibodies or antigens labeled with highly sensitive markers such as 111 isotopes, fluorescent substances, luminescent substances, or enzymes to form antigen-antibody complexes. Methods for detecting and analyzing test substances with high sensitivity have also been proposed. However, since the former method using fine particles is inferior in sensitivity to the latter method using markers, recently the latter method of analysis using markers with high detection sensitivity has become mainstream.

As analysis methods using such markers, radioimmunoassay using radioisotopes as markers, fluorescence immunoassay using fluorescent substances, enzyme immunoassay using enzymes, etc. are known. Among them, enzyme immunoassay has been attracting particular attention recently because it does not require special equipment or measurement techniques and can be easily performed using a commonly used colorimeter. This enzyme immunoassay is a homogeneous method that quantifies a test substance by directly determining the amount of change in the activity of a labeled enzyme depending on the presence or absence of an immunochemical reaction.
genetic) enzyme immunoassay and an insoluble carrier,
For example, using a synthetic resin such as plastic or glass beads, the enzyme-labeled antibody that has reacted with the antigen or antibody, or the enzyme-labeled antigen and the unreacted one, are separated into B-F by a washing operation, and the labeled enzyme after this B-F separation is separated. Heterogeneous (Heterogeneo), which quantifies the test substance by determining the amount of activity.
US) It is classified into two methods: enzyme immunoassay.

However, although the former homogeneous enzyme immunoassay method can be performed with simple operations, it has the disadvantage that it cannot analyze 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 separation, but cannot properly analyze whether the test substance is a low-molecular or high-molecular substance. , is becoming popular because its analysis targets are extremely wide-ranging.

As such heterogeneous enzyme immunoassay methods, competitive methods, Sand-Deutsch methods, and the like are known. In the competitive method, as shown in Figure 1, an antibody or antigen that causes an antigen-antibody reaction with a test substance in a sample is immobilized on an insoluble carrier 1 in advance, and this carrier 1, sample, and its test substance 2 are immobilized. The same substance is subjected to an antigen-antibody reaction with an enzyme-labeled labeled reagent 3, and then washed, and the antigen-antibody reaction competes with the carrier 1 and binds to the -C-bound test substance 2 a5 and the labeled reagent 3. B. After separating them, add a coloring reagent that reacts with the labeled enzyme in labeling reagent 3 to react, and then measure the reaction solution colorimetrically to determine the enzymatic activity of the labeled enzyme. It is used to quantify test substance 2.As shown in Figure 2, the Sandermansch method is similar to the competitive method in that the antibody or antigen is pre-immobilized without causing an antigen-antibody reaction with the test substance in the sample. Using an insoluble carrier 5, first an antigenic reaction is performed between the carrier 5 and the sample 7 to bind the test substance 6 in the sample to the carrier 5, and then washing is performed to separate B-F. The carrier 5 is treated with a labeled reagent 7, which is an enzyme-labeled substance that causes an antigen-antibody reaction with the test substance 6, to cause an antigen-antibody reaction, and then washed again to separate B-F, and then the labeled reagent 7 is added. After a coloring reagent that reacts with the labeled enzyme in 7 is added and reacted, the reaction solution is colorimetrically measured to determine the enzymatic activity of the labeled enzyme, and the amount of the test substance 6 is quantified.

As mentioned above, in the heterogeneous enzyme immunoassay method, in the competitive method, once during the analysis of one test substance;
In the sandwich method, two B-F separations are required, and when a reaction vessel for antigen-antibody reactions is to be used repeatedly, the reaction vessel must be This means that a cleaning process will be added. In this way, when automating the enzyme immunoassay method, which requires at least two washing steps including B-F separation for the analysis of one test substance, it is necessary to install a dedicated washing device for each washing step. Although it is possible to do so,
If this is done, the device becomes large, complicated, and expensive. Such problems similarly occur when automating the above-mentioned radioimmunoassay, fluorescence immunoassay, etc. that use markers.

SUMMARY OF THE INVENTION An object of the present invention is to solve the above-mentioned problems and to provide an automatic immunological analysis method that can be carried out using a compact, simple-configured, and inexpensive analyzer.

The present invention provides a carrier on which a predetermined antibody or antigen is immobilized;
When automatically immunologically analyzing a test substance in a sample by carrying out an antigen-antibody reaction in a reaction container using a predetermined antibody or a labeled reagent in which an antigen is labeled with a predetermined substance, the reaction container During the analysis of the test substance in the sample contained in the reaction vessel, the sample is transported at least twice to a washing device installed in the reaction line to remove the antibody or antigen bound to the carrier and the carrier. This method is characterized in that washing including B-F separation for separating uncontained antibodies or antigens is carried out at least twice.

The present invention will be described in detail below with reference to the drawings.

FIG. 3 is a diagram showing the configuration of an example of an enzyme immunoassay automatic analyzer according to the present invention, which employs the sandwich method shown in FIG. 2. In this example, a single reaction line is used to analyze a single item. The reaction container has a large opening 11a.
and using 24 O-shaped tubes 11 having small openings 11b,
These are held on the same circumference of the reaction tube disk 12 at equal intervals.

The reaction tube disk 12 is intermittently rotated at a predetermined pitch (for example, 15 seconds) in the direction indicated by the arrow in a horizontal plane while the O-shaped tube 11 is immersed in the constant temperature bath 10 (FIG. 4). The stopping positions of the O-shaped tube 11 due to the intermittent rotation of the reaction tube disk 12 are not indicated by symbols S1 to 824. In this example, a sample cup 1 is placed at a predetermined sample suction position of a sampler 14 by a sample dispensing device 13 into a zero-shaped tube 11 located at a stop position S1.
Selectively dispense the sample from step 5. In addition, sampler 1
4 is 2, which is the same number as the number of U-shaped tubes held in the reaction tube disk 12.
Four sample cups are held on the same circumference (at equal intervals) and rotated intermittently in the direction of the arrow in synchronization with the rotation of the reaction tube disk 12. '1
The reagent dispensing device 16 selectively dispenses the enzyme labeled reagent 17 according to the analyte in the sample, and moves to the stop position S4.
The coloring reagent 19 is selectively dispensed by the reagent dispensing device 18.
One insoluble carrier 21 such as a synthetic resin such as plastic or glass beads, which is housed in a large number in a carrier injector 20, is selectively introduced into the O-shaped tube 11 located at 7 from its large opening 11a. In addition, the carrier 21 is the large mouth part 11a of the 0-shaped tube 1.
It has a large size that can be easily taken in and taken out from the mouth part 11b and does not enter the small opening 11b, and has an antibody or an antigen that causes an antigen-antibody reaction with the test substance in the sample immobilized on its surface in advance as described above. The carrier injector 20 is kept moist with a buffer solution. Further, from the O-shaped tube 11 located at the stop position S+s, the reaction liquid contained therein is selectively sucked into the colorimeter 22, and from the O-shaped tube 11 located at the stop position 821, the reaction liquid accommodated therein is selectively drawn into the colorimeter 22. The carrier 21 is selectively taken out and discharged by a carrier extractor 23. Furthermore,
A cleaning pump 21I is attached to the O-shaped pipe 11 at the stop position 822.
At the stop position 824, the buffer solution 26 is selectively injected using the buffer solution dispensing device 25. Further, each of the O-shaped tubes 11 located at the stop positions 82 to S5 has its mouth portion 11b removably connected to the common stirring air pump 27, and similarly, the O-shaped tubes 11 located at the stop positions @522d3 and S23 are removably connected to the common stirring air pump 27. Each of the 0-shaped tubes 11 has its mouth portion 11b removably connected to a common drainage pump 28.

The present inventors solved the above-mentioned problems and developed a carrier on which a predetermined antibody or antigen is immobilized as an immunological automatic analysis method that can be carried out using a small, simple, and inexpensive analyzer Q. and a labeled reagent in which a predetermined antibody or antigen is labeled with a predetermined substance, and an antigen-antibody reaction is performed in a reaction container to automatically immunologically analyze a test substance in a sample. During the analysis of the test substance in the sample contained in the reaction vessel, the reaction vessel is circulated and conveyed to a cleaning device installed in an endless reaction line to remove antibodies or antigens bound to the carrier. and washing including B/F separation to separate antibodies or antigens not bound to the carrier.
We are proposing an automatic analysis method that can be carried out repeatedly. In addition, as an apparatus for carrying out such an automatic analysis method, a large number of reaction vessels are arranged circumferentially on a turntable, and the turntable rotates three or four times for each sample, thereby performing analysis. We also propose something like this. In such an automatic analyzer, after the turntable has made one revolution and dispensed all samples from the reaction vessels, the next sample is dispensed until the turntable has rotated two or three more times. This method has disadvantages in that sample dispensing becomes irregular, and not only does it become troublesome to control the drive timing of the sample dispenser, but it also becomes troublesome to control the ID and process the analysis results. Furthermore, if sample dispensing cannot be carried out continuously in this way, there is also the disadvantage that sample processing efficiency decreases.

It is an object of the present invention to eliminate these drawbacks and to provide a mechanism for cleaning the reaction vessel by circulating it through the same cleaning device during the analysis of each sample, while also allowing continuous dispensing of the sample. The purpose of this paper is to provide an automatic immunological analysis method.

The present invention provides a carrier on which a predetermined antibody or antigen is immobilized;
In automatically immunologically analyzing a test substance in a sample by performing an antigen-antibody reaction in a reaction container using a predetermined antibody or a labeled reagent in which an antigen is labeled with a predetermined substance, the reaction described above is performed. During the analysis of the test substance in the sample contained in the reaction container, the container is circulated to a cleaning device disposed in an endless reaction line to remove the carrier (antibodies or antigens bound thereto). and washing including B and F separation for separating antibodies or antigens not bound to the carrier are performed at least twice, and the sample is continuously dispensed into the anti-Luff container. be.

The present invention will be described in detail below with reference to the drawings.

FIG. 3 is a diagram showing the configuration of an example of an automatic enzyme immunoanalyzer for carrying out the method of the present invention, which employs the Sandersch method shown in FIG. 2G.

In this example, as an anti-J core container, a U having a large opening], la, and a small opening 1], b as shown in the fourth figure.
25 tubes 11 are used, and these are maintained at equal intervals of one circle around the same circumference of the reaction tube disk 12. While soaking in the water, hold it in the direction indicated by the arrow for a specified pinch (for example, 15 seconds).
Rotate intermittently. The stop positions of the U-shaped tube 11 due to the intermittent rotation of the reaction tube TIF 12 are indicated by symbols S to S25.
Indicated by In this example, the sample is selectively dispensed from the sample cup 15 at a predetermined sample suction position of the sampler 14 by the sample dispensing device 13 from the U-shaped tube 1.14 located at the stop position S. Note that any type of sampler 144 can be used, but in this example, a large number of racks 14.a each holding 10 sample cups are held side by side, and the rank of the left column is 3rd.
In the figure, the rank in the right column that holds the sample cup that has been dispensed is moved upward to be transported to the sample dispensing position by sequentially moving downward. The rack located at the sample dispensing position G is moved in synchronization with the rotation of the reaction tube disk 12 by the arrow S.
move intermittently in the direction of

JX (D Rack G) After dispensing all the held samples, this rack will be sent to the bottom of the rack row on the right, and the rack at the bottom of the left row will be the next sample dispensing position. In this way, successive samples can be continuously sent to the sample dispensing position G at a predetermined pitch.

U-shaped tube 11 at stop position S□ of opposite/main tube disk 12
Then, the first reagent dispensing device 16 selectively dispenses the Chiku 1 reagent 17. A buffer is used as this first reagent.

U-shaped tube 11 in stop position S3. &, : is the analyte Q in the sample by the second reagent dispensing device i″i J, 8.
The corresponding enzyme reagent 19 is selectively dispensed. Also, the U-shaped tube at the stop position S2], ]i is the third reagent dispensing device 208.
Selectively dispense coloring reagent 2 from this.

Further, a U-shaped tube with a stop position of 115" SIQ]-1, G is its large part], 1. From a to the carrier loading device 22
From G, one of the insoluble carriers 23 such as synthetic resins such as plastics and glass heath, which are stored in large numbers therein, is selectively introduced. Note that the carrier 23 can be easily taken in and out from the large opening hia of the U-shaped tube 11, and the small opening 1], b
The groove is so large that no one is needed, and as described above, an antibody or an antigen that causes an antigen-antibody reaction with the test substance in the sample is immobilized on its surface in advance. Also, stop position 5
From the 2oQ U-shaped tube 11-, the reaction liquid contained therein is used as a colorimetric device! t24. The carrier 23 accommodated therein is selectively taken out and discharged from the U-shaped tube 11 at the stop position @S23 by the carrier discharge device 25. Furthermore, the stop position S2. A cleaning device 26 selectively injects and discharges cleaning solutions such as ion-exchanged water, immunoassay buffer, and physiological saline into the U-shaped tube 11 for B/F separation and the U-shaped tube 11. Perform cleaning.

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

For example, the analysis is carried out by the Sand-Deutsch method, and for each sample, the reaction tube disk 12 rotates three times to complete the analysis.

In other words, B-F separation is performed twice (this U-shaped tube is used repeatedly and cleaning is performed once. Therefore, the sample is dispensed, and the first, second, and third reagents are separated). Dispensing,
Loading, discharging, and supplying the carrier 23 to the colorimeter are performed once while the reaction tube disk 12 moves three pitches. However, since cleaning is performed three times during analysis as described above, it is performed at each pitch of movement of the reaction tube disk 12. In addition, in order to operate in this way, the number of U-shaped tubes 11 loaded with the reaction tube disk 12G is 3n+1 or 3 when n is 1.2.3...
It is necessary to set it to n+2. In this example, the number of U-shaped tubes 11 is 25, which is 3n+1 (1=8).

During the first rotation of the reaction tube disk 12, first, one carrier 23 is placed in the stop position 81G, the U-shaped tube 11G, and the carrier input device 22 as shown in Figure 44.
”￥ii51.1. Insert from a. This stop position S
At the same time, 17 predetermined amounts of the first reagent consisting of a buffer solution are dispensed from the first reagent dispensing device @16G. This reaction tube 1
After the sample No. 1 is fed three pitches, the sample amount is dispensed from the sample dispensing device 13G at the stop position tffis4.

This initiates an antigen anti-f4 reaction. The reaction tube at the end of the first rotation G reaches the stop position SQ5, where it is cleaned by the cleaning device 26, and the first rotation B.
F separation is performed. In FIG. 5, the timing of the operation performed on the sample is indicated by diagonal lines downward to the left.

Next, the reaction tube disk 12 enters the second rotation, and at the stop position S3, the inside of the U-shaped tube 11 (second reagent dispensing device 1
8, a predetermined amount of the enzyme labeling reagent 19 is dispensed, and the second reaction is started. The final stop position S2 of this second rotation. A second B-F separation is performed by the cleaning device 25.

Furthermore, the reaction tube disk 12 enters the third rotation and stops at the stop position 82 (at this point, a third reagent dispensing device 2 is inserted into this U-shaped tube).
0 [From this, a predetermined amount of the coloring reagent 21 is dispensed, and the third reaction is started. When reaching stop position S26G, U-shaped tube 1
The test liquid in 1 is sucked by the pump of the colorimetric device 24 and guided to the colorimetric cell, where a colorimetric measurement using light of a predetermined wavelength is performed.

Next, by rotating three pitches, the carrier 23 remaining in the U-shaped tube is removed by the carrier discharging device 25 at the stop positions S and G. The final stop position S2 of the third rotation.

At this point, the U-tube 11 is cleaned by the cleaning device 26 and used repeatedly for analysis of the next sample. In FIG. 5, the operation timing for the European sample is indicated by diagonal lines downward to the right.

The cleaning device 26 cleans the large opening 11a of the U-shaped tube 11.
The cleaning liquid is intermittently injected in a shower-like manner, and the liquid is sucked and discharged from the small opening 11b using a drainage pump. 1. I can do something. As shown in Figure 4,
2G has a cleaning liquid tank 26a1 and a cleaning liquid supply pump 26.
A b1 nozzle 26C1 drainage pump 26d, etc. are provided. Further, the carrier feeding device 22 is also provided with a cage device 2zb for separating and supplying 11 pieces of carriers 23 from a hopper 22a1 which stores a large number of carriers 23, as shown in FIG. 14G. There is. Generally, the book 23 is held in the hopper 22a in a state moistened with a buffer solution. Furthermore, the carrier 11 discharge device j') 1.25 lowers the nozzle to the large opening VflI], ], aG, of the U-shaped tube 11, and removes the carrier H\23 by adsorbing it to the tip of the nozzle through the suction G. It can be lowered into the large opening of the U-shaped tube and taken out by grasping the carrier f4 23.

As described above, the analysis operation for one sample G is completed when the reaction tube disk J2 rotates three times, or in this example, the sample dispensing, the first, the second, and so on. The third reagent dispensing, loading and discharging the carrier, and colorimetric measurement are performed when the reaction tube disk 12 moves 8 pitches and operates once. For example, when the sample dispensing device 13 operates at the stop positions S and G, the U-shaped tubes C' are mounted at equal intervals. We will pass each other. This situation occurs about 1 for 3 binges.
Since it can be said that all the motions Q that are performed twice are correct, the sample portion 71 can be performed continuously G once every three pitches. In the past, it became possible to control samples and process analysis results at regular intervals.
, : , it becomes easy to control various kinds of sounds.

Furthermore, the U-shaped tube 11 is connected to one washing device 25 provided in the reaction line without an entrainment line.
Since the G filter is used to circulate and transport the B and F particles and to repeatedly perform washing including B and F separation, the main structure of the device is compact and has 11 sections.
This can be done at a low cost.

FIG. 6 shows the configuration of an example of an automatic analyzer for carrying out the immunological analysis method of the present invention using a competitive method.
The figure is a timing chart for explaining the operation. 6. In J4, the same components as those shown in FIG. 3G are indicated by the same symbols (=J. When performing this, add a sample and an enzyme labeling reagent to a carrier bound with a predetermined antigen or antibody, perform an antigen-anti-H reaction, perform B/F separation, and then: Since the colorimetric measurement is carried out after the reaction vessel has been prepared, two washes are required, including washing the reaction vessel.
In the example shown in FIG.
For reagent dispensing, loading/unloading of carriers, and colorimetric measurements, strain the reaction tube so that it operates once every time the tube is moved (12 or 2), and wash it for each tube (
If this is a problem, the sample can be dispensed continuously, ie every two steps G.

In Fig. 6 Q, the U-shaped tube 11 (the moss first dispensing neck J8
The first reagent 19 consisting of an enzyme labeling reagent is dispensed, and at the same time one carrier is introduced through the carrier input device 22. Further, a second reagent 21, which is a coloring reagent, is dispensed from the second reagent dispensing device 20 and the U-shaped tube 11 located at the stop position S2G. Furthermore, the U-shaped tube 11G is located at the stop position s3G, and the sample dispensing device L3+c is used to dispense the nozzle. In this example, there are a plurality of samples (7) 9 Sampler 1 holding a plurality of sample ranks 14 holding nozzle cups 15
4) G to supply the main position Q
Rub. The U-shaped tube 11, which is a reaction vessel, has a large opening, ia, and a small opening 11b in the lower right corner of FIG. The U-shaped tube 11 at the stop position S2° aspirates the test liquid into the colorimeter 2 and G, and returns to the stop position S24. Then, the carrier 23 in the U-shaped tube 11 is discharged from the carrier discharging device 25&, and the carrier 23 is moved to the stop position 16S2. Then, the cleaning device 26 cleans the U-shaped tube and performs B-F separation.

The operation timing of the automatic analyzer shown in FIG. 3 is shown in FIG. 7 (not shown). When one carrier 23 is introduced, the first reagent dispensing device 18 dispenses the enzyme labeled reagent 19.
Dispense a predetermined amount. When the reaction tube disk 12 moves two steps and the U-shaped tube 11 stops, i'& S
3 When you reach t
A predetermined amount t1'' of the sample is applied through 7'i J 3G, and an antigen-antibody reaction is performed. When this U-shaped tube 11 reaches the stop position S251, that is, at the last step position of one rotation Ll, the sample is removed from the cleaning device 26G. BF separation is performed.Next, after proceeding 2 steps, stop position 52
i = left number:? A predetermined amount L1 of a coloring reagent 2t, which is a second reagent, is dispensed from the sample 5-minute meter device 20G, and a second reaction is started. When this U-tube 11 reaches the stop position S2°O, the test liquid is drawn into the colorimetric cell from the colorimetric devices 2, 1, and Q, and color comparison 71!1] is carried out. (After this 2 step knob, the stop position iRS 24.
Discharge by step 5. 3rd rotation final stop position S 2
, B, the empty U-shaped tube 11 is cleaned by the cleaning device 26 &
The sample is then washed and prepared for the next analysis of Sample G. A predetermined analysis can be performed by applying IW to each sample (reaction tube disk 12) twice.In this example, 25 (2n+1) U-shaped tubes are installed in the reaction tube disk 12. Since the samples were arranged in such a way that sample dispensing, reagent dispensing, loading, discharging, and colorimetric measurements were performed once every two snaps, successive samples were continuously Can be dispensed.

However, the cleaning device 2G must be operated for each step (1, / Figure 8 shows another example of an automatic analyzer for carrying out the method of the present invention using the Sand Inch method). In this example as well, the parts shown in Figure 3 and times t71 and 2 are shown with the same symbols. In this example, the U-shaped tube 11 is fixedly arranged on the counter pressure tube 251.The difference between this example and the apparatus shown in FIG. Stop position 323~
The three IJ-shaped tubes in S25 are cleaned simultaneously (this is the point mentioned above). By configuring the cleaning device 2G in this way, it is no longer necessary to operate the cleaning device 2G at each step. As with other operating mechanisms, it is preferable to operate the G reactor tube disk once every three steps of 2.The drive control for these operations can be made common.Also, with such a configuration Therefore, the carrier discharging device 25 is provided at the stop position WS20.

FIG. 9 shows the operation timing of the automatic analyzer shown in FIG. 8. As you can see, the mounting in this example is almost the same as the one shown in Figure 8, and requires three cleaning steps.
Since there are only two differences in this one time, the explanation thereof will be omitted. With this f14 configuration, the sample can be continuously dispensed every three steps.

FIG. 10 shows still another example of an automatic analyzer for carrying out the analysis method according to the present invention, and in this example, enzyme immunoassay is performed by the sandwich method. In the above-mentioned example, one row of U-shaped tubes 11 was arranged on the reaction tube disk 2, but in this example, three rows of reaction tubes 32 are provided on the reaction disk 31.

Each row has 247 reaction tubes, or this number ("
The responsibility is. For convenience of explanation, the outermost reaction tube row is referred to as the second reaction tube row 32-]-1, and the middle reaction tube row is referred to as the second reaction tube row 32-2. The innermost identical reaction tube row is connected to the third reaction tube row 32-
It is called 3. As in Example 011, the anti-core tube disk 31 moves intermittently (twice) at a predetermined pitch. At the stop position SIG, the carrier feeding device 33 is set to transfer the carrier to the reaction tube (
Input this selectively. In this example, this carrier loading device 33 is the first carrier loading device 33. G can be supplied sequentially to the second and third anti-J heart tube rows.

Zunawaji, first reaction tube row 32-10 1 in the reaction tube of normal flow
For the small number of carriers that were introduced one by one, carriers were introduced one by one into successive reaction tubes in the second reaction tube row 32-2, and then one by one into successive reaction tubes in the third reaction tube row 32-3. Inject the January body and repeat the reaction in sequence in the first reaction tube row 32-1 again.
-=--=-LM ← ← Fried ≠ Odd bones G The carrier is introduced one by one, and thereafter the same G is applied and the carrier introduction operation is repeated.

Stop position 82G moss cleaning device ↑Soil 31. is provided so that all reaction tubes 32 at this stop position are simultaneously cleaned. A first reagent dispensing device @35 is placed at the stop position S8, and a first reagent 36 made of a buffer solution is dispensed into the reaction tube 32. This partial pressure is also the same as the carrier charging device 33. The first reagent is dispensed in the order of the second and third reaction tube rows. 4.0 Stop position S-port is sample dispensing device @
37 is provided to partially pressure the samples sequentially supplied by the sampler 38 into the reaction tube 82. This sample dispensing is also performed for each successive reaction tube row. A second reagent dispensing device 39 is provided at the stop position S5, and a second reagent 4, which is an enzyme-labeled reagent, is disposed at the stop position S5.
・Dispense 0. At the sixth stop position 860, the moss third reagent dispensing device 4.1 is installed, and the third reagent 4.1 is an enzyme coloring reagent.
Dispense 2. These second and third reagent dispensing are also performed for each successive reaction tube row.

Colorimetric devices 4 and 8 are provided at the stop position @S23, and the test liquids in the reaction tubes 32 of the sequential reaction tube array are guided to the colorimetric cell for colorimetric measurement. Also, at the stop position @S24, there is a carrier ejecting device 4.
, 4. are provided to discharge the carrier remaining in the reaction tube. These colorimetric devices 4, 3 and 4JH body discharge devices 4, 4, and I
It operates for each row of reaction tubes. 10th
In the figure, there are three
The connections between other mechanisms and reaction tubes are shown by one solid line and two broken lines.
It was shown that this works.

1] is a diagram showing the operation timing of the self-analyzing device shown in FIG. .

32-2 and 32-3.

The first cleaning is performed at a timing synchronized with the reaction tube TIF31.
-G is carried out at the same time in the third reaction tube row.

First, one reaction tube (carrier) of the first reaction tube row 32-1 is introduced at the stop position 81. This reaction tube is cleaned at the next stop position S2. Before cleaning, this reaction tube is Is there any test liquid remaining in the previous sample?
This test solution does not contain any substances that would bind to the carrier, so there is no contamination of the carrier. However, by washing with stop position GS, contamination between the previous test solution and the sample or reagent can be avoided. Nation will also disappear. Next, at the stop position S3, the first reagent dispensing device 35 dispenses a predetermined amount of the first reagent 36 made of a buffer solution into the reaction tube. When it moves one more step and reaches the stop position S, a predetermined amount of sample is dispensed into this reaction tube by the sample dispensing device 37. reaction is initiated. In this example, the steps are as follows: loading the carrier, washing, dispensing the first reagent, dispensing the sample, first reaction column, sequential reaction column (in contrast, sequential reactions, that is, one step at a time). .

When the second reaction tube of the first reaction tube row into which the first sample has been dispensed reaches the stop position S2 again, the first B-F separation is performed by the cleaning device 34.G.

This reaction tube is at this stop position S2. (The process of reaching this point (here, the stopping position S5 + S6 + S2'3 + S
24. As is clear from FIG. 11, there is no hindrance because these are operating against the reaction tubes G of the second and third reaction tube rows. Next, when this reaction tube reaches the stop position S5, the second reagent dispensing device 39
A predetermined amount of the second reagent 40 is dispensed, and the second reaction is started. When this reaction tube is further sent to the stop position S2, it is washed again and a second B-F separation is performed.

Thereafter, at the stop position S6, a predetermined amount of the third reagent 42 is dispensed by the third reagent dispensing device 41, and the third reaction is started. When this reaction tube is further sent to the stop position S23, colorimetric measurement is performed by the colorimetric devices 4 and 3G. Next, at the stop position S24, the carrier remaining in the reaction tube is discharged from the reaction tube by the carrier discharge device 4.4°. Up to this point, the reaction tube has been rotated in the forward direction, and when it moves one step and enters the stop position S, the carrier is thrown in again and the above-mentioned operation is repeated.

On the other hand, regarding the carrier charging device 33, in the first rotation, jl of the first reaction tube row 32-1 [1
Once the carriers have been introduced into all 24 reaction tubes, the process moves to the second reaction tube row 82-2.
One by one, the carriers were introduced into 4□ reaction tubes, and then the third
Moving to the reaction tube row 32-3, carriers are sequentially introduced into the 241 reaction tubes one by one. Next, the first anti-element, tube row 32
The process moves to -1, and the operations described in -1- are not repeated. Sample dispensing device 37, first reagent dispensing device 35, three second reagent dispensing devices, third reagent dispensing device 41, colorimetric measurement device 4
・The operation of J3 and carrier discharge device 4.1 is also similar,
As shown in FIG. 11, the target reaction tube rows are different. For example, in the first rotation in FIG. 11, carrier injection is performed to the first reaction tube row 32-1, first reagent dispensing is performed from the third step to the first reaction tube row, and For the 2nd trial mulberry dispensing, the second reaction tube row 32- is used until the 4th step.
2, and from the fifth step to the third reaction tube row 3
Moving on to 2-3, the dispensing of the third reagent is performed to the first reaction tube row G in the fifth step 1, and is performed to the second reaction tube row 32-2 from the sixth step onward (・In this way, in this example, sample dispensing can be performed continuously for each step of the reaction tube 31, so the sample processing efficiency is improved when compared with the above-mentioned example. Furthermore, since the loading and unloading of carriers, sample dispensing, reagent dispensing, and colorimetric measurements are performed for each reaction tube row in sequence, these driving controls become extremely easy.

The present invention is not limited to the embodiments described above, but can be modified in many ways. In the above example, enzyme immunoassay using an enzyme-labeled reagent is performed, but it can also be applied to number-pair immunoassay using a number-pair isotope as a marker, fluorescence immunoassay using a fluorescent substance as a marker, etc. can do. Further, the reaction tube does not necessarily have to be held on a disk-shaped reaction tube tray, and for example, a snake chain or gondola type conveying device can be used. Furthermore, in the above example, the final test solution was introduced into the colorimetric cell (colorimetric measurement), but a transparent reaction tube was used, and the colorimetric measurement was performed while the test solution was in the reaction tube (in the state where it was present). It is also possible to adopt a direct photometry method that performs the following.

In this case, if the carrier remaining in the reaction tube interferes with photometry, the carrier can be removed before photometry. Also, when using such a direct metering method (here,
Since the carrier can be discharged together with the test solution after photometry, the carrier discharge device becomes simple. Furthermore, although one cleaning device is provided in the two-way embodiment, a plurality of cleaning devices may be provided. For example, in the embodiment shown in FIG.
A second cleaning device can also be provided at an opposite location. Even in this case, compared to the case where three cleaning devices are provided, the device can be made simpler and more compact. Further, in the above-described embodiments, the reaction tubes are used repeatedly, but this is not always necessary, and the reaction tubes used for analysis may be disposable. Furthermore, in the above-described embodiments, all samples are analyzed for the same measurement item, or multiple items may be analyzed at the same time.

Furthermore, various dispensing positions, carrier loading and discharging positions, colorimetric measurement positions, etc. are not limited to those of the above-mentioned embodiment G, and various changes can be made. Further, although the example described above does not mention anything about stirring, a suitable stirring mechanism can be provided at a suitable stop position. For example, when a U-shaped reaction tube is used, stirring can be achieved by supplying air from the small opening of the tube.

As explained above, in the automatic immunological analysis method of the present invention, during the analysis of each sample, the reaction tube is circulated through a cleaning device installed in the reaction line configured with an endless gas line, and the sample is Since the system is designed to perform continuous dispensing, the overall configuration of the automatic analyzer can be made simple and compact.Moreover, the operation of all mechanisms, including sample dispensing, can be controlled at a common timing, making control easier. becomes easier. It is possible to easily control the sample ID and process the separation results, which is the advantage of 4L'.

[Brief explanation of the drawing]

Figure 1 shows enzyme immunoassay due to competition u < i
Figure 2 (-) shows the process of enzyme immunoassay using the sandwich method (χ1, Figure 13 shows an automatic analyzer rjσ that carries out the analysis method according to the present invention) - Configuration of an example Figure 4 is a diagram that cleverly shows the operation of Ill<j order, Figure 5 is a timing diagram showing the operation of each part, and Figure 6 is an analysis of the present invention. ) A diagram showing the configuration of another example of an automatic analyzer line that implements the following; FIG. 7 is a timing diagram for explaining its operation; FIG. 8 is an analysis method of the present invention. FIG. 9 is a diagram showing the configuration of still another example of an automatic analyzer for carrying out the analysis, FIG. 9 is a timing chart diagram for explaining its operation, and FIG. 40 is an automatic analyzer for carrying out the analysis method of the present invention. FIG. 11 is a timing diagram for explaining the operation of the structure of still another example. Dispensing device 144・Sampler】5 Sample cup 1.6, 18, 20・Reagent dispensing device 22-・Carrier loading device 23・Carrier 2.1・Colorimetric device 25・・Carrier discharging device 2G・Washing device
31. Reaction tube 1 entrance tube disk 32... Reaction treatment 2 32-1, 32-2, 32-3 Reverse J heart tube row 83
...Carrier loading device 34.・Cleaning device 35.39
．． J・]・Reagent dispensing device, 37・・Sample dispensing device 38・・Zumpura 4・3・
Colorimetric device 4,44...Carrier discharge device. Procedural amendment 1 March 4, 1988 ■, Minor case indication 1988 Patent Application No. 9599 2, Title of invention Immunological automatic analysis method 3, Person making the amendment Relationship with the case Special remarks' 1 Applicant (037) Olympus Optical Industry Co., Ltd. 5゜6, Subject of amendment Claims in the specification, Detailed description of the invention σ) d Patent on page 1, lines 3 to 17 of the specification letter e
Correct the range of ft calculation as follows. [2. Claim L: A sample is prepared by performing an antigen-antibody reaction in a reaction container using a carrier on which a predetermined antibody or antigen is immobilized and a labeling reagent in which the predetermined antibody or antigen is labeled with a predetermined substance. In automatically immunologically analyzing the test substance contained in the reaction vessel, the reaction vessel is installed in an endless reaction line during the analysis of the test substance in the sample contained in the reaction vessel. Circulate and transport to cleaning equipment
Washing including B-F separation for separating the antibody or antigen bound to the carrier from the antibody or antigen not bound to the carrier is performed at least twice, and the sample is continuously dispensed into the reaction container. An automated immunological analysis method characterized by: ”

Claims (1)

[Claims] 1. Using a carrier on which a predetermined antibody or antigen is immobilized and a labeling reagent in which the predetermined antibody or antigen is labeled with a predetermined substance, an antigen-antibody reaction is performed in a reaction container to prepare a sample. In automatically immunologically analyzing the test substance contained in the reaction vessel, the reaction vessel is installed in an endless reaction line during the analysis of the test substance in the sample contained in the reaction vessel. The sample is circulated to a washing device and washed at least twice, including B-F separation to separate antibodies or antigens bound to the carrier from antibodies or antigens not bound to the carrier, and the sample is transferred to a reaction vessel. An automatic immunological analysis method characterized by continuously dispensing.