JPS61118662A - Immunological automatic analysis - Google Patents

Abstract

PURPOSE:To efficiently perform the replacement of a carrier and a marker reagent in order to change over an analytical item, by displaying that each of plural analytical operations was finished with respect to the predetermined number of samples. CONSTITUTION:In a carrier charging control apparatus 8, the charging number of the carriers 17 received in a carrier charging device 16 are counted and, when the counted value became equal to the number of the samples set to a sampler 14, a display apparatus 35 displays that the charging of the carriers with respect to all samples to the analytical item concerned was finished on the basis of said information. Similarly, in a nozzle transfer control apparatus 32, the distribution number of times of a first reagent 26, a second reagent 28 and a third reagent 30 are counted and, when the counted value to each reagent became equal to the number of the sample set to the sampler 14, the display apparatus 3 displays that the distribution of the reagents with respect to all samples to the analytical item concerned was finished on the basis of said information.

Description

Detailed Description of the Invention (Technical Field) The present invention relates to an automatic immunological analysis method.0 (Prior Art) In recent years, with advances in medical care, it has become possible to analyze minute amounts of biological components, and It is useful for early diagnosis, etc. For example, intractable diseases include malignant tumors represented by α-fetoprotein and carcinoembryonic antigen, disorders of abnormal secretion of hormones represented by insulin and thyroxine, and immune diseases represented by immunoglobulins. Not only can it be used to diagnose various diseases at an early stage, but also to monitor these diseases after treatment, and recently, low molecular weight haptens (such as drugs) can be used.
It has also become possible to measure incomplete antigens, which is also useful in creating drug administration plans.

Many of these biological components are analyzed by immunochemical methods that utilize antigen-antibody reactions. An antigen-antibody reaction is carried out using an insoluble carrier such as a synthetic resin or glass beads, and a labeling reagent in which a specified antibody or antigen is labeled with a highly sensitive marker such as a radioactive isotope, a fluorescent substance, or an enzyme. B-
Petrogenic, which separates B' and quantifies the analyte in the sample based on the labeled reagent after B-F separation.
(Heterogeneous) immunoassay method is al.

The Kono immunoassay method is becoming popular because it can properly analyze whether the test substance is a low molecule or a high molecule, and it can be analyzed over a very wide range of subjects.

As such heterogeneous immunoassay methods, the competitive method, the sandwich method, etc. are known. As shown in Figure 1, in the competitive method, an antibody or antigen that causes an antigen-antibody reaction with a test substance in a sample is immobilized in advance on an insoluble carrier,
This carrier 1 is subjected to an antigen-antibody reaction with a labeling reagent 8 that is enzyme-labeled 11 on the same substance as the sample and its test substance 2, and then washed, and the antigen-antibody reaction competitively binds to the carrier 1. Test substance 2 and labeled reagent 8
and those that are not bound are separated by B-F, then a coloring reagent that reacts with the labeled enzyme in labeling reagent B is added and reacted, and the reaction solution is measured colorimetrically. The test substance 2 is quantified by determining the enzyme activity of the labeled enzyme.

As shown in Figure 2, the Sand Inch method uses an insoluble carrier 5 on which an antibody or antigen that causes an antigen-antibody reaction with the test substance in the sample is preliminarily immobilized, similar to the competitive method. First, an antigen-antibody reaction is performed between the carrier 1 and the sample to bind the test substance 6 in the sample to the carrier 5.
Next, after washing and B-F separation, the carrier 5 is treated with a labeling reagent 7 labeled with a substance that causes an antigen-antibody reaction with the test substance 6, for example, with an enzyme, to cause an antigen-antibody reaction. After washing again and separating B-F, a coloring reagent that reacts with the labeling enzyme in labeling reagent 7 is added and reacted, and the reaction solution is colorimetrically measured to determine the enzyme activity of the labeling enzyme. This is to measure a fixed amount of test substance 6. In addition,
In the sandwich method, the carrier 5, sample and labeling reagent 7 may be reacted simultaneously; in this case, B-
F separation will be performed once.

The applicant has developed an apparatus for carrying out such a heterogeneous immunoassay method, but as is clear from the above explanation, carriers, labeling reagents, etc. are not suitable for the 0-in-1 analysis. This varies depending on the substance to be tested. Therefore, when developing such an automatic analyzer, it is better to make it possible to exchange carriers, labeling reagents, etc., so that analysis can be performed for each analysis item. This is advantageous in terms of simplification and miniaturization of the device, ease of control, etc. In this case, in order to efficiently analyze multiple items, it is necessary to completely complete the analysis of a certain item on multiple samples. After that, carrier, labeling reagent, etc.
Rather than replacing it with the one used for the next analysis item,
It is preferable to replace the predetermined number of samples after the necessary analysis operations have been completed.

(Object of the invention) In view of the above points, the object of the invention is to efficiently exchange carriers, labeled reagents, etc. for switching analysis items, and therefore to efficiently perform analysis of multiple items. The purpose of this study is to provide an automatic immunological analysis method that can be used.

(Summary of the Invention) The present invention provides a carrier on which a predetermined antibody or antigen is immobilized;
A predetermined antibody or antigen is labeled with a predetermined substance using a labeling reagent, and the antigen and antibody are combined in a reaction container.

Immunology...
In automatic analysis, each of multiple analysis operations, such as adding a carrier corresponding to the analyte to each reaction container and dispensing a reagent, is performed for a predetermined number of samples. This feature is characterized by displaying that each process has been completed.

(Example) FIG. 8 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 sandwich method shown in FIG. 2.

In this example, 24 U-shaped tubes 11 each having a large opening 11a and a low opening 11b are used as reaction vessels, and these are held at equal intervals on the same circumference of the reaction tube disk 12. The reaction tube disk 12 connects the U-shaped tube 11 to a constant temperature bath.
While immersing it in water, it is intermittently rotated at a predetermined pitch (for example, 15 seconds) in the direction indicated by the arrow in a horizontal plane. The stopping positions of the U-shaped tube 11 due to the intermittent rotation of the reaction tube disk 12 are indicated by symbols 81 to ``g4.'' Sample 2・□ at the predetermined sample suction position in U14
Selectively dispense the sample from cup 15. Oh, I
The sampler 14 holds 24 sample cups, the same number as the number of U-shaped tubes held on the reaction tube disk 12, at equal intervals on the same circumference, and intermittently moves in the direction of the arrow in synchronization with the rotation of the reaction tube disk 12. Rotate it. - Insoluble carriers 17 such as synthetic resins such as plastics and glass beads, which are stored in large numbers in the carrier injector 16, are fed into the U-shaped tube 11 at the stop position S17 from its large opening 11a under the control of the carrier injecting control device 18. Insert one piece selectively. The carrier 17 has a size that allows it to be easily taken in and taken out from the large opening 11a of the U-shaped tube 1...11, but does not enter the small opening 11b, and the surface thereof contains the test substance and antigen antibody in the sample as described above. The antibody or antigen that causes the reaction is immobilized in advance, and the carrier injector 16 is moistened with buffer 1.

In addition, from the U-shaped tube 11 located at the stop position S19, the reaction liquid contained therein is selectively sucked into the colorimeter 19,
From the U-shaped tube 11 at the stop position S28, the carrier 17 housed therein is selectively taken out by the carrier extractor 20! ···Discharge. Furthermore, the cleaning pump 21 selectively injects a cleaning liquid such as ion-exchanged water, a swab solution for immunoassay, or physiological saline into the U-shaped tube 111 at the stop position S22.

Further, a stirring air pump 2z is detachably connected to the small end 11b of the U-shaped tube 11 at the stop position S, and the U-shaped tube 11 is also moved to the stop position S8. A common drainage pump 28 is removably connected to the small opening 11b of each U-shaped tube 11 in S28.

Furthermore, the U-shaped tube 11 located at the stop position S24 has...
A syringe-type reagent dispenser 24 capable of precision dispensing allows the first reagent (buffer) 26 in the first reagent container 25, the second reagent (enzyme labeled reagent) 28 in the second reagent container 27, and the eighth reagent to be dispensed.
One portion of any one of the eighth reagents (coloring reagents) 80 in the reagent container 29 is poured through the nozzle 81. This nozzle 81
is transferred by the nozzle transfer control device 82 to the stop position "'84" and the suction position of each reagent and the cleaning tank 84 via the nozzle transfer device 88.
d.
- In this example, the carrier input control device 18 counts the number of carriers stored in the carrier input device 16, and when the counted value becomes equal to the number of samples set in the sampler 14, Based on this information, the display device 85 displays that carrier loading has been completed for all samples for the relevant analysis item. Similarly, the nozzle transfer control device 82 counts the number of times each of the first reagent 26, the second reagent 28, and the eighth reagent 80 is dispensed, and the count value for each reagent is calculated based on the sample & I set in the sampler 14.
When it becomes equal to ll, based on that information, the display device 86 displays that the dispensing of the reagent has been completed for all samples for the analysis item. This display device 85
As the light source, a known one such as a CRT or an LED is used.

Next, the operation of the enzyme immunoassay automatic analyzer shown in FIG. 8 will be explained. First, the first to
Set the eighth reagent containers ff15, 17, and 119, respectively. Further, the number of sample cups set in the sampler 14, ie, the number of 2゛□ samples to be analyzed, is input in advance to the carrier input control device 18 and the nozzle transfer control device 88. During the first rotation of the reaction tube disk 18, the carrier 17 corresponding to the analysis item is first introduced into the U-shaped tube 11 by the carrier injector 16 at the stop position S11. Carrier 1
7 is inserted into the U-shaped tube 11, it is at the stop position S2. At a point a, the cleaning pump 21 and the drain pump 28 are operated to cleanse, and at the stop position S□, a fixed amount of the first reagent 26 is dispensed by the reagent dispenser 24. After that, the stop position S0
By the operation of the sample dispensing device 18, a certain amount of sample is dispensed from the sample cup 15 at a predetermined sample suction position, and at the next stop position S8, it is stirred by the operation of the stirring air pump 22. Reaction 1 begins. The above carrier loading operation, first reagent dispensing operation, and sample.

After the dispensing operation has been performed a number of times equal to the number of samples set in the sampler 14, these operations are not performed.

At the 1u turn of the reaction tube disk 1s, the carrier injector 16 injects the carrier 17 a number of times equal to the number of samples set in the sampler 14.

When the operation is finished, the display device 86 displays the first message, and similarly, when the reagent dispenser 24 finishes dispensing the first reagent 26 a number of times equal to the number of samples set, the first reagent 26 is dispensed. At that point, a message to that effect is displayed on the display device 85.

During the second rotation of the reaction tube disk 12, at the stop position 8o, the U
The double tube 11 is washed and the first B-F separation is performed, and then a fixed amount of the second reagent 28 is injected by the reagent dispenser 24 at the stop position S□, and the second reaction begins. . The test solution in the U-shaped tube 11 into which the second reagent 28 has been dispensed is stirred by the stirring air pump 22 at the stop position S. At the second rotation of the reaction tube disk 12, one reagent is dispensed. The number of samples for which the device 24 is set is 1
When the dispensing operation of the second reagent 28 is completed a number of times equal to
At that point, a message to that effect is displayed on the display device 85.

At the eighth rotation of the reaction tube disk 12, the U-shaped tube 11 is cleaned by the action of the cleaning pump 21 and the drain pump 28''' at the stop position S, and the second IB-F separation is performed, and then it is stopped. Reagent dispenser 24 at position "84"
The nozzle transfer device @84 selects the coloring reagent container 29, dispenses a certain amount of the eighth reagent 80, and starts the eighth reaction.The eighth reagent 80 is dispensed into the U-shaped tube 11. The test solution is stirred at the stop position S to promote the enzyme reaction. At the eighth rotation of the reaction tube disk 12, the reagent dispenser 24 is rotated a number of times equal to the number of samples set.
When the dispensing operation of the reagent 80 is completed, a message to that effect is displayed on the display device 85 at point 1''.

Thereafter, at the fourth rotation of the reaction tube disk 12, the test liquid in the U-shaped tube 11 is sucked into the colorimeter 19 at the stop position 819 and measured colorimetrically, and then at the stop position S, the U-shaped tube 1, The carrier 1 remaining in the U-shaped tube 11 is taken out by the 1" carrier extractor 20. After that, the U-shaped tube 11 is cleaned by the cleaning pump 21 and the drain pump 28 at the stop position S, and then the U-shaped tube 11 is removed from the next stop. At position Ss8, the remaining liquid is drained by the drain pump 28 and prepared for the next analysis.
During the above operation, the nozzle 81 is in the cleaning tank after dispensing each reagent.
184, and the inner and outer walls of the nozzle 81 are cleaned by suction and discharge operations of the reagent dispenser 24.

As described above, in this example, the carrier 1, the 1st reagent 26, and the 1st reagent 26, which need to be replaced depending on the analysis item
, the second reagent 28, and the eighth reagent 80, the fact that the input operation and dispensing operation have been completed for the set sample is displayed at the end of each operation. You can efficiently exchange and set up carriers and reagents to be used in the next analysis item from those that have been completed. As a result, multiple items can be analyzed efficiently for multiple samples.

Note that the present invention is not limited to the above-mentioned example, and can be modified or modified in many ways. For example, in the above-mentioned embodiment, the number of operations was counted, but if An appropriate identification member is provided at the last sample cup 15 or in the vicinity of its set position, and this is detected by, for example, an optical detection means, and the completion of each operation for the final sample is determined based on the detection signal of the SC. Table 1 may also be used. In addition, in the above-mentioned examples, enzyme immunoassay was carried out using the Sand-Deutsch method, but it can be similarly applied to analysis using a competitive method. It can be similarly applied to fluorescence immunoassay using optical substances. Furthermore, the reaction container does not necessarily need to be held on a disk; for example, a snake chain or gondola type conveying device can also be used.

However, the sampler can also be changed in various ways, such as by using a rack. Furthermore, in the above example, the finally obtained test solution was introduced into the colorimetric cell for colorimetric measurement, but a transparent reaction container is used and the colorimetric measurement is performed with the test solution present in the reaction container. A direct photometry method can also be adopted. In this case, if the carrier remaining in the reaction vessel interferes with photometry, the carrier can be removed before photometry.

In addition, when using such a direct metering method, 1
Since the carrier can be discharged together with the test solution after photometry, the carrier discharge device becomes simple. Further, in the above-described embodiment 1, the reaction container is used repeatedly, but this is not always necessary, and the reaction container used for analysis can be made disposable. Further, the various dispensing positions, carrier loading and discharging positions, colorimetric side positioning, etc. are not limited to the above-mentioned embodiments, and various changes are possible.

(Effects of the Invention) As described above, in the automatic immunological analysis method of the present invention, multiple analysis operations such as loading carriers and dispensing reagents corresponding to analysis items into each reaction container are performed. Since each item is displayed to indicate that the predetermined number of samples has been completed, it is possible to efficiently exchange and set carriers, various reagents, etc. for switching analysis items. Therefore, analysis of multiple items on multiple samples can be performed efficiently.

[Brief explanation of drawings]

Figure 1 is a diagram showing the process of enzyme immunoassay using the competitive method.
FIG. 2 is a diagram showing one process of enzyme immunoassay using the Sand-Deutsch method, and FIG. 8 is a diagram showing the configuration of an example of an automatic analyzer for carrying out the analysis method according to the present invention. 11...U-shaped tube 12...Reaction tube disk -18...Sample dispensing device @14...Sampler 15...Sample cup 16...Carrier injector 17...Carrier 18... - Carrier input control device 19... Colorimeter 20... Carrier extractor 21... Washing pump 22... Stirring air pump 28... Drainage pump 24... Reagent dispenser 2
5...First reagent container 26 River first reagent 27...
Second reagent container 28... Second reagent 29... Eighth reagent container 80... Eighth reagent 81... Nozzle 82... Nozzle transfer control device 88... Nozzle transfer device 8 cleaning tank
185...display device

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 detect the test substance in the sample. In automatic immunological analysis, each of multiple analysis operations, such as loading carriers corresponding to the test substance into each reaction container and dispensing reagents, is performed based on a predetermined number of samples. 1. An automatic immunological analysis method, characterized in that each item is displayed as having been completed.