JPH0577981B2 - - Google Patents

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, malignant tumors represented by α-fetoprotein and carcinoembryonic antigen, diseases of abnormal secretion of hormones represented by insulin and thyroxine, etc., and immune diseases represented by immunoglobulins, etc., are considered to be incurable diseases. It is not only possible to diagnose various diseases at an early stage, but it is also useful for monitoring those diseases after treatment, and recently, it has become possible to measure low-molecular haptens (incomplete antigens) such as drugs, which is useful for formulating 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. How to analyze test substances,
Radioactive isotopes, fluorescent substances,
A method of analyzing a test substance by detecting an antigen-antibody complex with high sensitivity using a labeled antibody or a labeled antigen labeled with a highly sensitive marker such as a luminescent substance or an enzyme has 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.

Analysis methods using such markers include:
Known methods include radioimmunoassay using a radioisotope as a marker, fluorescence immunoassay using a fluorescent substance, and enzyme immunoassay using an enzyme.
Among these, 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 method is divided into a homogeneous enzyme immunoassay method, in which the test substance is quantified by directly determining the amount of change in the activity of a labeled enzyme based on the presence or absence of an immunochemical reaction, and a homogeneous enzyme immunoassay method, in which the test substance is quantified by directly determining the amount of change in the activity of a labeled enzyme, depending on the presence or absence of an immunochemical reaction, and the other is a homogeneous enzyme immunoassay method, in which the test substance is quantified by directly determining the amount of change in the activity of a labeled enzyme, depending on the presence or absence of an immunochemical reaction. The enzyme-labeled antibody that has reacted with the antigen or antibody, or the unreacted enzyme-labeled antigen and the unreacted one, are separated into B and F by a washing operation using synthetic resins and glass beads, and the activity amount of the labeled enzyme after this B and F separation is determined. It is classified into two methods: heterogeneous enzyme immunoassay, which determines the amount of the test substance and quantifies it. However, although the former homogeneous enzyme immunoassay method can be performed with simple operations, it has the disadvantage that it can only 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 to perform B/F separation, but it can properly analyze whether the test substance is a low-molecular or high-molecular one.
It 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, insoluble carrier 1
An antibody or an antigen that causes an antigen-antibody reaction with a test substance in a sample is immobilized in advance, and an antigen-antibody reaction is performed between the carrier 1, the sample, the test substance 2, and a labeling reagent 3 which is an enzyme-labeled substance that is the same as the test substance 2. After that, washing is performed, and the test substance 2 and labeling reagent 3 are competitively bound to the carrier 1 by an antigen-antibody reaction.
After separating the unbound and unbound substances by B and F, a coloring reagent that reacts with the labeled enzyme in labeling reagent 3 is added and reacted, and the reaction solution is measured colorimetrically to determine the enzymatic activity of the labeled enzyme. The test substance 2 is quantified by determining the following. In addition, as shown in FIG. 2, in the Sanderutsch method, like the competitive method, an insoluble carrier 5 on which an antibody or antigen that causes an antigen-antibody reaction with the test substance in the sample is immobilized in advance is used. The test substance 6 in the sample is bound to the carrier 5 by performing an antigen-antibody reaction with the sample, and after washing and separating B and F, the test substance 6 and the antigen-antibody are transferred to the carrier 5. A substance that causes a reaction is reacted with a labeling reagent 7 labeled with an enzyme to cause an antigen-antibody reaction, and then washed again to separate B and F, and then a coloring reagent that reacts with the labeling enzyme in the labeling reagent 7 is added. After addition and reaction, the reaction solution is subjected to colorimetric measurement to determine the enzyme activity of the labeled enzyme, and the amount of the test substance 6 is quantified.

As mentioned above, in the heterogeneous enzyme immunoassay method, during the analysis of one test substance, B/F separation is required once in the competitive method and twice in the Sand-Deutsch method, and antigen-antibody reactions are not performed. When a reaction vessel is used repeatedly, an additional step is required to wash the reaction vessel after the analysis of one sample is completed and before the analysis of the next sample is started. In this way, when automating the enzyme immunoassay method, which requires at least two washing steps including B and F separation for the analysis of one test substance, it is necessary to carry out a dedicated washing process for each washing step. Although it is conceivable to arrange the device, this method has the problem of making the device large, complicated, and expensive. Such problems similarly occur when automating the above-mentioned radioimmunoassay, fluorescence immunoassay, etc. that use markers.

The present inventors solved the above-mentioned problems and developed an automated immunological analysis method that can be carried out using a compact, simple-configured, and inexpensive analyzer using a carrier immobilized with a predetermined antibody or antigen, In automatically immunologically analyzing a test substance in a sample by performing an antigen-antibody reaction in a reaction container using a labeled reagent in which an antibody or an antigen is labeled with a predetermined substance, the reaction container is During the analysis of the test substance in the sample contained in the reaction vessel, the antibody or antigen bound to the carrier and the antibody or antigen bound to the carrier are circulated and transported to a cleaning device installed in the endless reaction line. We have proposed an automatic analysis method in which washing, including B and F separation to separate uncontaminated antibodies or antigens, is performed at least twice. 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 each sample is analyzed by rotating the turntable three or four times. We are also proposing something that looks like this. In such an automatic analyzer,
After the turntable has made one revolution and the sample has been dispensed into all reaction vessels, the next sample cannot be dispensed until the turntable has rotated two or three more times, which may result in sample dispensing being incomplete. This has the disadvantage that not only is it difficult to control the drive timing of the sample dispenser, but also it is difficult to control the ID and process the analysis results. Also, if sample dispensing cannot be performed continuously like this,
Another disadvantage is that sample processing efficiency is reduced.

The purpose of the present invention is to eliminate such drawbacks, to complete the analysis by circulating the reaction vessels multiple times during the analysis of each sample, and to continuously perform sample dispensing to multiple reaction vessels even during this circulation transfer. The purpose of the present invention is to provide an automatic immunological analysis method that can be performed automatically.

In the automatic immunological analysis method of the present invention, a sample to be analyzed and a labeled reagent in which a predetermined antibody or antigen is labeled with a predetermined substance are added to a solid-phase reagent in which a predetermined antibody or antigen is immobilized. After performing an antigen-antibody reaction in a reaction container, the test substance in the sample is washed for B/F separation to remove substances that have not bound to the immobilized reagent. For automatic analysis, multiple reaction vessels are conveyed multiple times along an endless reaction line, and samples are dispensed into each of a predetermined number of reaction vessels. The dispensing of the sample to all the reaction vessels is completed only after the sample is transferred, and the dispensing of labeled reagent and/or washing for B/F separation to the reaction vessels from which the sample has been dispensed is carried out through the circulation. This method is characterized in that it is performed during transportation and during a different rotation from the sample dispensing.

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 Sanderuch method shown in FIG. 2.
In this example, the reaction vessel is a U having a large opening 11a and a small opening 11b as clearly shown in FIG.
25 tubes 11 are used, and these are connected to the reaction tube disk 1.
2 at equal intervals on the same circumference. The reaction tube disk 12 is intermittently rotated at a predetermined pitch (eg, 15 seconds) in the direction shown by the arrow while the U-shaped tube 11 is immersed in the constant temperature bath 10 (FIG. 4). The stopping positions of the U-shaped tube 11 due to the intermittent rotation of the reaction tube disk 12 are indicated by symbols S ₁ to S ₂₅ . 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 into the U-shaped tube 11 at the stop position _S4 . Although any type of sampler 14 can be used, in this example, a large number of racks 14a each holding 10 sample cups are used.
The racks in the left column are sequentially moved downward in FIG. 3 and transported to the sample dispensing position, and the racks in the right column, which hold the sample cups that have been dispensed, are moved upward. The rack at the sample dispensing position is intermittently moved in the direction of arrow S in synchronization with the rotation of the reaction tube disk 12.
When all the samples held in this rack have been dispensed, this rack is sent to the bottom of the rack row on the right, and the rack at the bottom of the left row is sent next to the sample dispensing position. . In this manner, successive samples can be continuously delivered to the sample dispensing position at predetermined pitches.

The first reagent 1 is supplied to the U-shaped tube 11 at the stop position S ₁ of the reaction tube disk 12 by the first reagent dispensing device 16.
Selectively dispense 7. A buffer is used as this first reagent. The second reagent dispensing device 18 selectively dispenses an enzyme labeled reagent 19 corresponding to the analyte in the sample into the U-shaped tube 11 located at the stop position _S3 . Further, the coloring reagent 21 is selectively dispensed into the U-shaped tube 11 at the stop position _S2 by the third reagent dispensing device 20. Furthermore, one carrier 23 of synthetic resin such as plastic or insoluble carrier such as glass beads is selected from the large opening 11a of the U-shaped tube 11 located at the stop position _S1 using the carrier feeding device 22 from the large opening 11a. to invest in. The carrier 23 has a size that allows it to be easily taken in and taken out from the large opening 11a of the U-shaped tube 11 and does not need to enter the small opening 11b. The antibody or antigen is immobilized in advance. In addition, the reaction liquid contained therein is transferred from the U-shaped tube 11 located at the stop position S ₂₀ to the colorimetric device 24.
U-tube 1 in stop position S ₂₃
1, the carrier 23 accommodated therein is selectively taken out and discharged by a carrier discharge device 25.
Furthermore, a cleaning device 26 selectively injects and discharges cleaning solutions such as ion exchange water, immunoassay buffer, and physiological saline into the U-shaped tube 11 located at the stop position S ₂₅ to perform B/F separation and U-tube 11. The cross tube 11 is cleaned.

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

In this example, analysis is performed by the Sand-Deutsch method, and for each sample, the reaction tube disk 12 rotates three times to complete the analysis. That is, B and F separation is performed twice and cleaning is performed once for repeated use of the U-shaped tube. Therefore, dispensing the sample, dispensing the first, second, and third reagents, loading and discharging the carrier 23,
The supply to the colorimeter etc. is operated once during the movement of the reaction tube disk 12 by three pitches. However, since cleaning is performed three times during the analysis as described above, it is performed for each moving pitch of the reaction tube disk 12. In addition, in order to operate in this way, the U loaded in the reaction tube disk 12 must be
The number of double tubes 11 is when n is 1, 2, 3...
It must be 3n+1 or 3n+2. In this example, the number of U-shaped tubes 11 is 25, which is 3n+1 (n=8).

In the first rotation of the reaction tube disk 12, one carrier 23 is first introduced into the U-shaped tube 11 at the stop position _S1 from the carrier input device 22 through its large opening 11a, as shown in FIG. At this stop position _S1 , the first reagent dispensing device 16 simultaneously dispenses a predetermined amount of the first reagent 17 made of a buffer solution. After this reaction tube 11 is fed three pitches, a predetermined amount of sample is dispensed by the sample dispensing device 13 at the stop position _S4 . This initiates an antigen-antibody reaction. At the end of the first rotation, this reaction tube is at the stop position S ₂₅
At this point, cleaning is performed by the cleaning device 26, and the first B/F separation is performed. Fifth
In the figure, the timing of the operation performed on the sample is indicated by diagonal lines going downward to the left.

Next, the reaction tube disk 12 enters the second rotation, and at the stop position _S3, the second reagent dispensing device 18 dispenses a predetermined amount of the enzyme labeled reagent 19 into the U-shaped tube 11, and the second reaction starts. be done. At the last stop position _S25 of this second rotation, the second B/F separation is performed by the cleaning device 25.

Furthermore, the reaction tube disk 12 enters the third rotation,
At the stop position _S2 , a predetermined amount of the coloring reagent 21 is dispensed into this U-shaped tube by the third reagent dispensing device 20, and the third reaction is started. When the stop position _S20 is reached, the test liquid in the U-shaped tube 11 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, when it rotates 3 pitches, at the stop position _S23 , the carrier discharging device 25 removes the carrier 2 remaining in the U-shaped tube.
Remove 3. At the final stop position S ₂₅ of the third rotation, 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 next sample is indicated by a diagonal line downward to the right.

Cleaning by the cleaning device 26 can be carried out by intermittently injecting a cleaning liquid in a shower-like manner from the large opening 11a of the U-shaped tube 11 and suctioning and discharging it from the small opening 11b using a drainage pump. As shown in FIG. 4, the cleaning device 26 is provided with a cleaning liquid tank 26a, a cleaning liquid supply pump 26b, a nozzle 26c, a drainage pump 26d, and the like. Further, as shown in FIG. 4, the carrier loading device 22 also includes a hopper 22a for storing a large number of carriers 23, and a hopper 22a for storing a large number of carriers 23.
Gate device 22b that separates and supplies 3 one by one
etc. are provided. Generally, the carrier 23 is held in the hopper 22a in a state moistened with a buffer solution. Further, the carrier discharging device 25 lowers the nozzle to the large opening 11a of the U-shaped tube 11, and removes the carrier 23 by adsorbing it to the tip of the nozzle by suction, or lowers the arm into the large opening of the U-shaped tube to remove the carrier 23. You can grab it and take it out.

As described above, the analysis operation for one sample is completed when the reaction tube disk 12 rotates three times, but in this example, sample dispensing, first, second, and third reagent dispensing, and carrier dispensing are performed. input, discharge,
For colorimetric measurement, the reaction tube disk 12 moves 3 pitches and operates once, and the reaction tube disk 12 moves 25 pitches.
Since (3×8+1) U-shaped tubes 11 are installed at equal intervals, for example, when the sample dispensing device 13 is operated at the stop position S ₄ , the U-shaped tubes positioned are one rotation of the reaction tube disk 12. It will be shifted by one item each time. Since this situation applies to all operations performed once for every three pitches, sample dispensing can be performed continuously once every three pitches. Therefore, the sample
ID control and processing of analysis results can now be performed at regular intervals, making various controls easier. Furthermore, the reaction line is endless,
One cleaning device 25 provided in the reaction line includes a U
Since the tubular tube 11 is circulated and carried out to repeatedly perform cleaning including B/F separation, the overall structure of the apparatus can be made small and simple, and can be made at 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, and FIG. 7 is a timing chart for explaining its operation. In FIG. 6, the same components as those shown in FIG. 3 are designated by the same reference numerals. As explained with reference to Fig. 1, when performing immunological analysis using a competitive method, the sample and enzyme labeling reagent are added to a carrier to which a predetermined antigen or antibody is bound, and an antigen-antibody reaction is performed.・F
Since separation is performed, then an enzyme coloring reagent is added, and colorimetric measurements are performed, washing is performed twice, including washing the reaction vessel. Therefore, in the example shown in FIG. 6, 2n+1 U-shaped tubes 11 are arranged on the reaction tube disk 12, and sample dispensing, reagent dispensing, loading/unloading of carriers, colorimetric measurements, etc. are carried out on the reaction tube disk 12. If the operation is performed once every two steps, and washing is performed after each step, the sample can be dispensed continuously, that is, every two steps.

In FIG. 6, a first reagent 19 consisting of an enzyme-labeled reagent is dispensed by a first dispensing device 18 into the U-shaped tube 11 located at a stop position S ₁ on the circumference of the reaction tube disk 12, and a carrier charging device 1 by 22
Insert. In addition, the U-shaped pipe 11 at the stop position _S2
A second reagent 21, which is a coloring reagent, is dispensed by the second reagent dispensing device 20. Further, a sample is dispensed by a sample dispensing device 13 into the U-shaped tube 11 located at the stop position _S3 . In this example as well, the sample is sequentially supplied to the sample dispensing position from the sampler 14 having a plurality of sample racks 14a holding a plurality of sample cups 15. The U-shaped tube 11, which is a reaction vessel, has a large opening 11a and a small opening 11b, as shown in the lower right corner of FIG. The test liquid is sucked into the colorimeter 24 from the U-shaped tube 11 at the stop position _S22 , the carrier 23 in the U-shaped tube 11 is discharged by the carrier discharge device 25 at the stop position _S24 , and the carrier 23 is discharged from the U-shaped tube 11 at the stop position _S25. The cleaning device 26 cleans the U-shaped tube and performs B and F separation.

FIG. 7 shows the operation timing of the automatic analyzer shown in FIG. 6. Looking at the analysis operation for a certain sample, first, one carrier 2 is loaded into the U-shaped tube 11 by the carrier loading device 22 at the stop position _S1 .
At the same time, a predetermined amount of the enzyme labeled reagent 19 is dispensed by the first reagent dispensing device 18. When the reaction tube disk 12 moves two steps and the U-shaped tube 11 reaches the stop position _S3 , a predetermined amount of sample is dispensed by the sample dispensing device 13, and an antigen-antibody reaction is performed. When this U-shaped tube 11 reaches the stop position _S25 , that is, at the last step position of the first rotation, the cleaning device 26 performs B and F separation. Next, after proceeding two steps, the second reagent dispensing device 20 dispenses a predetermined amount of the coloring reagent 21, which is the second reagent, at the stop position _S2 , and the second reaction is started. When the U-tube 11 reaches the stop position _S22 , the test liquid is sucked into the colorimetric cell by the colorimetric device 24 and colorimetrically measured. After two more steps, the carrier 23 remaining in the U-shaped tube 11 is discharged by the carrier discharge device 25 at the stop position _S24 . At the last stop position _S25 of the second rotation, the empty U-shaped tube 11 is cleaned by the cleaning device 26,
Prepare for analysis on the next sample. In this way, two reaction tube disks 12 are prepared for each sample.
By rotating it, a predetermined analysis can be performed. In addition, in this example, 25
(2n+1) U-shaped tubes 11 are arranged, and sample dispensing, reagent dispensing, carrier loading, discharging, and colorimetric measurement are performed once every two steps, so that sequential samples are collected at a cycle of two steps. Continuous dispensing is possible. However, the cleaning device 26 must operate for each step.

FIG. 8 shows another example of an automatic analyzer for carrying out the method of the present invention using the Sandersch method, and in this example as well, the same parts as shown in FIG. 3 are given the same reference numerals. If the reaction vessel is used repeatedly in the Sand-Deutsch method, it must be washed 3 times.
In this example, 25 U-shaped tubes 11 are arranged on the reaction tube disk 12 because it is necessary to repeat the reaction several times. The difference between this example and the device shown in FIG. 3 is that in this example, the cleaning device 26 simultaneously cleans the three U-shaped tubes 11 located at the stop positions S ₂₃ to _{S 25} . be. With this configuration, it is no longer necessary to operate the cleaning device 26 for each step, and it is sufficient to operate it once for every three steps of the reaction tube disk 12, similar to other operating mechanisms. Therefore, the drive control of these operating mechanisms can be made common. Furthermore, because of this configuration, the carrier discharge device 25 is provided at the stop position _S20 .

FIG. 9 shows the operation timing of the automatic analyzer shown in FIG. 8. As mentioned above, the apparatus of this example is almost the same as that shown in FIG. 3, and the only difference is that cleaning is performed once every three steps, so a description thereof will be omitted. Even with such a 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 Sand-Deutsch method. In the above example, reaction tube disk 1
In this example, three rows of reaction tubes 32 are provided on the reaction tube disk 31. Each row is provided with 24 reaction tubes, but this number is arbitrary. For convenience of explanation, the outermost reaction tube row is referred to as the first reaction tube row 32-1, the middle reaction tube row is referred to as the second reaction tube row 32-2, and the innermost reaction tube row is referred to as the third reaction tube row.
It is called a reaction tube row 32-3. As in the previous example, the reaction tube disk 31 is assumed to rotate intermittently at a predetermined pitch. A carrier loading device 33 is located at the stop position _S1 .
is provided to selectively charge the carrier into the reaction tube. In this example, this carrier loading device 33 is used for the first, second and third carrier loading devices.
It is designed so that it can be sequentially supplied to a row of reaction tubes. That is, the carriers are introduced one by one into the successive reaction tubes of the first reaction tube row 32-1, and then the carriers are introduced one by one into the successive reaction tubes of the second reaction tube row 32-2. Furthermore, carriers are introduced one by one into successive reaction tubes in the third reaction tube row 32-3, and again one by one into successive reaction tubes in the first reaction tube row 32-1. The carrier loading operation is repeated.

A cleaning device 34 is provided at the stop position _S2 to simultaneously clean all reaction tubes 32 at this stop position. The first reagent dispensing device 3 is located at the stop position _S3 .
5 and dispenses a first reagent 36 consisting of a buffer solution into the reaction tube 32. This dispensing is also carried out by the carrier injection device 3.
3, the first reagent is dispensed in the order of the first, second and third reaction tube rows. A sample dispensing device 37 is provided at the fourth stop position S ₄ and dispenses the samples sequentially supplied by the sampler 38 into the reaction tube 32 . 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 dispenses a second reagent 40, which is an enzyme-labeled reagent. A third reagent dispensing device 41 is provided at the sixth stop position _S6 to dispense a third reagent 42, which is an enzyme coloring reagent. These second and third reagent dispensing are also performed for each successive reaction tube row.

A colorimetric device 43 is provided at the stop position _S23 , and the test liquids in the reaction tubes 32 of the successive reaction tube rows are guided to a colorimetric cell for colorimetric measurement. Further, a carrier discharging device 44 is provided at the stop position _S24 to discharge the carrier remaining in the reaction tube. These colorimetric device 43 and carrier discharge device 44 are also operated for each successive reaction tube row. In FIG. 10, a cleaning device 3 that simultaneously cleans the reaction tubes of the first to third reaction tube rows is shown.
4 and the reaction tube are shown connected by three solid lines,
The other mechanisms and the reaction tubes are indicated by one solid line and two broken lines, indicating that they operate for each successive row of reaction tubes.

FIG. 11 is a diagram showing the operation timing of the automatic analyzer shown in FIG.
2-2 and 32-3. Cleaning is performed simultaneously on the first to third reaction tube rows at a timing synchronized with the reaction tube disk 31.

First, at the stop position _S1 , the first reaction tube row 32-1
One carrier is put into the reaction tube. The reaction tube is cleaned in the next stop position _S2 . Before washing, the test solution for the previous sample remains in this reaction tube, but since this test solution does not contain any substances that would bind to the carrier, the carrier may be contaminated. There is no need to stop at the stop position.
Washing with _S2 also eliminates contamination between the previous test solution and the sample or reagent.
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 _S4 , a predetermined amount of sample is dispensed into this reaction tube by the sample dispensing device 37, and the first reaction is started. In this example, such loading of the carrier, washing, dispensing of the first reagent, and dispensing of the sample are performed sequentially to successive reaction tubes in the first reaction tube array, that is, in each step.

The first tube of the first reaction tube row into which the first sample was dispensed
When the reaction tube reaches the stop position _S2 again, the cleaning device 34 performs the first B/F separation.
In the process of reaching this stop position _S24 , this reaction tube passes through stop positions _S5 , _S6 , _S23 , _S24 , _S1 , but as is clear from FIG. Since the operation is performed on the reaction tubes in the reaction tube row, there is no problem. Next, when this reaction tube reaches the stop position _S5 , a predetermined amount of the second reagent 40 is dispensed by the second reagent dispensing device 39, and the second reaction is started. This reaction tube is further sent to the stop position _S2 , where it is washed again and a second B/F separation is performed. Thereafter, a predetermined amount of the third reagent 42 is dispensed by the third reagent dispensing device 41 at the stop position _S6 ,
A third reaction is initiated. When this reaction tube is further sent to a stop position _S23 , a colorimetric measurement is performed by a colorimetric device 43. Next, at the stop position _S24 , the carrier remaining in the reaction tube is discharged from the reaction tube by the carrier discharge device 44. Up to this point, the reaction tube has rotated three times, and when it moves one step and enters the stop position _S1 , the carrier is introduced again and the above-mentioned operation is repeated.

On the other hand, regarding the carrier charging device 33, during the first rotation, carriers are charged one by one into the successive reaction tubes of the first reaction tube row 32-1, and carriers are charged into all 24 reaction tubes. When finished, the second reaction tube row 32-
2, one carrier is sequentially charged into each of the 24 reaction tubes, and then the process moves to the third reaction tube row 32-3, and one carrier is sequentially introduced into each of the 24 reaction tubes.
Next, the process moves to the first reaction tube row 32-1 again, and the above-described operation is repeated. Sample dispensing device 3
7, first reagent dispensing device 35, second reagent dispensing device 3
9, third reagent dispensing device 41, colorimetric measurement device 43,
The operation of the carrier discharging device 44 is also similar, but the 11th
As shown in the figure, the target reaction tube rows are different. For example, in the first rotation in FIG. 11, the carrier is introduced into the first reaction tube row 32-1,
The first reagent dispensing is performed to the first reaction tube array from the third step, the second reagent dispensing is performed to the second reaction tube array 32-2 up to the fourth step, and from the fifth step Moving to the third reaction tube row 32-3, the third reagent is dispensed to the first reaction tube row up to the fifth step, and from the sixth step onward to the second reaction tube row 32-2. It will be done. In this manner, in this embodiment, sample dispensing can be performed continuously for each step of the reaction tube disk 31, so that the sample processing efficiency is improved when compared with the previously described embodiment. In addition, 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-mentioned embodiment, enzyme immunoassay using an enzyme-labeled reagent is performed, but the present invention can be similarly applied to radioimmunoassay using a radioactive isotope as a marker, fluorescent immunoassay using a fluorescent substance as a marker, etc. I can do it. Further, the reaction tubes do not necessarily have to be held on a disk-shaped reaction tube disk, and for example, a snake chain or gondola type conveying device can be used. Furthermore, in the above example, the finally obtained test solution was introduced into the colorimetric cell for colorimetric measurement, but a transparent reaction tube was used and the colorimetric measurement was performed with the test solution present in the reaction tube. It is also possible to adopt a direct photometry method. In this case, if the carrier remaining in the reaction tube interferes with photometry, the carrier can be removed before photometry. Further, when such a direct photometry method is adopted, the carrier can be discharged together with the test liquid after photometry, so the carrier discharge device becomes simple. 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, a second cleaning device may be provided at a position substantially diametrically opposite the cleaning device 26. 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.
Further, in the above-described embodiment, all samples are analyzed for the same measurement item, but it is also possible to perform analysis for multiple items at the same time. Furthermore, various dispensing positions, carrier injection,
The discharge position, colorimetric measurement position, etc. are not limited to the above-mentioned embodiments, and various changes are possible. Furthermore, although no mention is made of stirring in the above example, a suitable stirring mechanism can be provided at a suitable stopping 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 automated immunological analysis method of the present invention, each reaction container is circulated multiple times along an endless reaction line during analysis of each sample, and multiple reactions on the reaction line are carried out. Since samples can be continuously dispensed into containers, the overall configuration of the automatic analyzer can be made simple and compact, and the sample processing efficiency is improved. Furthermore, since the operations of all mechanisms related to analysis including sample dispensing can be controlled at a common timing, control of the entire apparatus becomes easy.

[Brief explanation of the drawing]

Fig. 1 is a diagram showing the process of enzyme immunoassay using the competitive method, Fig. 2 is a diagram showing the process of enzyme immunoassay using the Sand-Deutsch method, and Fig. 3 is a diagram showing the process of enzyme immunoassay using the Sand-Deutsch method. A diagram showing the configuration of an example, FIG. 4 is a diagram showing its sequential operation, FIG. 5 is a timing chart showing the operation of each part, and FIG. 6 is an automatic analysis that implements the analysis method of the present invention. A diagram showing the configuration of another example of the device,
FIG. 7 is a timing chart for explaining its operation, FIG. 8 is a line diagram showing the configuration of still another example of an automatic analyzer that implements the analysis method of the present invention, and FIG. 9 is a diagram for explaining its operation. FIG. 10 is a diagram showing the configuration of still another example of an automatic analyzer that implements the analysis method of the present invention, and FIG. 11 is a timing chart diagram for explaining its operation. It is. 11...U-shaped tube, 12...reaction tube disk, 13...
sample dispensing device, 14... sampler, 15... sample cup, 16, 18, 20... reagent dispensing device,
22... Carrier input device, 23... Carrier, 24... Colorimetric device, 25... Carrier discharge device, 26... Washing device, 31
...Reaction tube disk, 32...Reaction tube, 32-1,3
2-2, 32-3... Reaction tube array, 33... Carrier loading device, 34... Washing device, 35, 39, 41... Reagent dispensing device, 37... Sample dispensing device, 38... Sampler, 43... Colorimetric device , 44...Carrier discharge device.

Claims (1)

[Scope of Claims] 1. A sample to be analyzed and a labeling reagent in which a predetermined antibody or antigen is labeled with a predetermined substance are placed in a reaction container with respect to an immobilized reagent in which a predetermined antibody or antigen is immobilized. After performing an antigen-antibody reaction, washing is performed for B/F separation to remove substances that have not bound to the immobilized reagent, and the test substance in the sample is automatically immunologically analyzed. In order to carry out the analysis, the plurality of reaction containers are transported multiple times along an endless reaction line, and the sample is circulated and transported multiple times by dispensing the sample into each of a plurality of predetermined reaction containers. The dispensing of the sample into all the reaction vessels is completed only after the dispensing of the sample is completed, and the dispensing of the labeling reagent and/or the washing for B/F separation to the reaction vessels in which the sample has been dispensed are carried out during the circulating transportation and An automatic immunological analysis method, characterized in that the method is carried out during a rotation different from the dispensing of the sample.