JPS5984159A - Method and device for automatic immune measurement of enzyme - Google Patents

Abstract

PURPOSE:To improve the efficiency in analysis treatment by making effective use of each idle treatment section while various reactions are caused with one sample, and performing the treatment of the other sample in said trearment sections in the idle time. CONSTITUTION:A titled means is so constituted that plural vessels or sets of plural vessels are successively accepted and pooled in a reaction zone 7 such as a reaction rotor and that the vessel or the set 1 of the vessel requiring any of the treatments by a washer 10, a reagent dispenser 11, etc. is subjected to the respective treatment stages by the control of a control device 8 consisting of a computer, etc., irrespectively of order in starting primary reaction, in the stage of immune measurement of enzyme. Thus, the idle sections are utilized effectively irrespective of idle sections in which samples are not fed, and treatment speed is improved considerably. The measurements for plural items are easily and securely executed simultaneously and parallel, a quick measurement is made possible without standing by for the arrangement of the specimens.

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to a method and apparatus for automatically performing enzyme immunoassay, and in particular, to a method and apparatus for automatically performing enzyme immunoassay, and in particular, to provide a reaction zone, which can be called a special section, in an analysis system, and to divide the reaction time equally. The present invention relates to a method and apparatus that enable a significant increase in processing speed.

Recently, in the field of clinical testing, analytical measurements using the enzyme immunoassay method (EIA method), which uses antigen-antibody reactions and enzyme reactions, have become popular. The currently commonly used EIA method has the disadvantage that the measurement operation is complicated and time consuming, which places a great burden on the inspector, and is inefficient.

In other words, the currently commonly used EIA measurement processes include the tube method and bead method depending on the object to which the antibody (or antigen) is immobilized, and the sandwich method and competition method depending on the reaction method. Taking the Sand Inch method as an example, it is as follows. First, place the sample (subject n￥) into a container (dupe) that has an antibody (or antigen) immobilized on its inner surface.
and bring it into contact and react (-next reaction), ■After the reaction is complete, B
-Wash for the F part, ■ Add enzyme labeling reagent and react (secondary reaction), ■ Wash again, ■ Add substrate reagent and perform enzyme reaction (tertiary reaction), ■ Add reaction stop reagent. In addition, each reaction requires a procedure of optically measuring the colored liquid caused by the enzymatic reaction and determining the amount of antigen (or antibody) in the sample from the fingernail line. This is time-consuming and patient work.

Therefore, in order to reduce the burden on inspectors, recently E'I A
2.3 A device for determining Δ111 internally and concavely has been developed (Japanese Patent Application Laid-open No. 57-174662, Japanese Patent Application Laid-open No. 56-14
7067 etc.) 01. (1) The first and second apparatuses follow the pre-L procedure as they are, and move containers (test tubes, G-cells) in the order in which they are stored, and perform various treatments and reactions during the movement.

Therefore, in these conventional devices, samples are received and analyzed in the order in which they are received, so it takes 11 hours to automatically analyze a large number of samples sequentially according to the same analysis process for a single item, but the reaction time is To measure multiple items with different processes, analyze each time 11! ! It is necessary to adjust the equipment so that the processing procedure and reaction time are compatible with the items. In addition, in some cases, it is not possible to change the order of samples to be provided later, even though the measurement should be completed first due to differences in the analysis process such as reaction time.
Measurement is performed one by one starting from the previous item (therefore, measurement can only be done after the previous item has been completed, and measuring 111 items requires a lot of time and is very inefficient).

In addition, in the conventional technology, the sample is sent in a fixed manner (speed) from the inlet to the outlet of the device, and the time required for analysis, such as reaction time, is determined by the item 'p sample. When the speed is increased, the distance t that the sample moves becomes longer and the size of the choc is increased.

Therefore, in the past, the sample transfer speed was set to be slower than the speed at which the sample could actually be delivered, and the sample dispensing mechanism introduced the sample into the apparatus in accordance with that speed.

Therefore, even if they follow the same analytical process,
If there is a blank space in the middle where the sample is not delivered, the device as a whole has a blank space (excess capacity) that can receive the sample, but the sample is delivered quickly enough to fill the blank space. However, since processing proceeds sequentially according to the sample introduction mechanism of the device, it is impossible to use those blank spaces later, and in this case, as mentioned above, the processing speed is very inefficient. (1ε become a thing.

Therefore, the present inventors conducted various studies to solve the above-mentioned drawbacks and improve the efficiency of automatic EIA measurements. The present invention was developed based on the fact that the time required for the reaction (described below) is significantly shorter than the time required for the reactions at each stage. That is, since the processing section is vacant while various reactions are being performed on one sample, efficiency is improved by processing other samples during this free time.

To this end, the system is divided into an analysis process and a reaction process, and the analysis process is divided into cleaning, dispensing, transfer of carriers (in the case of beads) and liquids in containers, and measurement processes, and the measurement items are divided into the following processes: We have adopted a system in which sample containers that require processing are sequentially provided with the corresponding processing step number, regardless of whether they are the same or different.
Note that this timing is automatically determined for each measurement item once the start time of the primary reaction, that is, the start time of contact is determined.

In addition, the equipment for washing, reagent dispensing, and measurement were made multifunctional to enable parallel analysis of multiple items.

Hereinafter, the present invention will be explained in detail based on embodiments shown in the drawings.

Figures 1 and 2 show an example of an automatic EIA measuring device based on a combination of the tube method and the sandwich method. Charger (5), cleaning device (11), reagent dispensing device (
11), a measuring device (control device JRC8, which is provided with 1→ and controls the operation of each device and performs various storage and calculations), a display/recording device 01, and the like. In addition, in this example, a reaction rack (1) as shown in FIG. Multiple reaction holes (4) provided in
Each one (14 in the figure) serves as a container. However, each independent container may be placed on a support such as a rack.
It is also possible to use a type that includes t. Furthermore, an antibody (or antigen) is immobilized on the inner surface of the reaction hole (4) marked with -, and measurement items (corresponding to the immobilized substance) are displayed on the rack. 4.1 The author may perform the display at the time of measurement.

Next, EIA i! by the device of the present invention. ! The I+ determination procedure will be explained along with the analytical processing process for one reaction rack.0 First, a sample is manually injected into each sample cup +3) on the turntable (2)L. reaction rack (
1) in the sample dilution/dispensing device (5), and
) Record the item name, number of samples, specimen number, etc. using the key (6a). The timing of cleaning the newly registered reaction rack (1), dispensing a certain amount of each fIV test, and measuring the start time does not overlap with the processing timing of each rack that is already being analyzed in the reaction rotor (7). It is selected by the control device (8), and the reaction part (reaction zone) is set to 11' when the reception is OK.
If there is a reaction rack storage groove available for the reaction rotor ((1)
is called, the sample is automatically diluted and dispensed into each reaction hole (4) of the reaction rack, and then automatically drawn into the accommodation groove (0) of the reaction rotor. At this time, try fi'
The start time (contact start time) of the primary reaction when l is added to the reaction hole (4) is determined by the control device (8 ). In addition, sample dilution dispensing% fi (
5) to the reaction rotor (7), various methods can be adopted, but in this example, a mechanism such as the fourth eye is used. 0 mating pin assemblies fixed to the rack transfer bell) (+4) and the motor (m) connected to the gear (+71), the control device (
When a signal to "transfer the reaction rack" is received from 8), the fitting pin (@ rotates and fits into the notch member of the reaction rack (1), and then the motor 0 frame rotates and the transfer belt (
The 1st member places the reaction rack (1) into the accommodation groove of the reaction rotor (7) (
). When the transfer is completed, the motor (frame 1) temporarily stops, the pin (A) comes off the notch (11), the motor (+8) reverses, the transfer belt (1→ returns to its original position, and the pin (A) is removed from the notch (11). ) is also returned to its original state and waits for the next reaction rack insertion.

When a predetermined reaction time for the primary reaction has elapsed in the reaction zone, the reaction rotor (7) rotates to transfer the reaction rack (1) to the cleaning device θ1. At this time, the reaction rack (1) is automatically pulled into the cleaning position from the reaction rotor (7), and predetermined cleaning is performed. The same number of cleaning nozzles (10a) are arranged in parallel as the number of reaction holes (4), and cleaning liquid is injected into and discharged from each reaction hole (4). After cleaning, the reaction rack automatically returns to its original position on the reaction rotor.

Next, the reaction rotor (7) rotates and transfers the reaction rack (]) to the reagent dispensing device 11). The reaction rack (1) is automatically drawn into the dispensing position, and the enzyme I'M identification reagent is dispensed in a predetermined amount. Enzyme labeled reagent dispensing nozzle (lla)
... are prepared in parallel for each measurement item, and which nozzle is activated is automatically determined by a command from the control device (8). Addition of reagents is done using a cough nozzle (
],]a) When moving the reaction rack (1) under the reaction rack (1), one inspection is performed for each reaction hole (4)... After dispensing the reagents, the reaction rack C1) automatically returns to its original position on the rotor. In addition, the reaction rotor (7) and cleaning device (■1,
The movement of the reaction rack between the reagent dispensing devices (11) is also IF of the sample dilution dispensing device (5)! Fit RIIi similar to i fit
It will be held in Then, the washing soil is placed (+(%) after the transfer is completed, and the reagent dispensing device (n) is transferred while it is being transferred, but in the latter case, an intermittent transfer method may be adopted. , When the predetermined process is completed, the motor reverses, transfers the reaction rack (1) to the original storage groove of the reaction rotor (7), and stops. (From 1!%) in the evening, it returns to its original position and prepares for the next transfer.

Now, after the reaction time for the reaction (secondary reaction) using this enzyme-labeled reagent has elapsed, the reaction rack (1) is subjected to the washing step as described above. Next, the reaction rotor (7) rotates and the reaction rack (1) is transferred again to the reagent dispensing device (11). The reaction rank (1) is automatically drawn into the dispensing position, and the enzyme substrate reagent is then dispensed into each reaction hole (4). When the enzyme substrate reagent dispensing nozzle (llb) is used in common for each measurement item as a labeled enzyme, one nozzle can be used for each measurement item. Addition of reagents is performed in the same manner as described above. After dispensing the reagents, the reaction rack (1) is automatically returned to its original position 1 lv on the reaction rotor.
Return to f.

When the reaction time of the enzymatic reaction (tertiary reaction) elapses, the reaction,
Rank (1) is also transferred to the reagent dispensing device (Ltl).

The reaction rack (1) is automatically pulled into the dispensing position, and this time the reaction stop reagent is dispensed. Reaction termination test: , I, I
The i-dispensing nozzle (llc) also has -
One nozzle can be used for 41 units using Rn enzyme.

Addition of reagents is performed in the same manner as described above. After dispensing the reagents, the reaction rack (1) automatically returns to its original position on the reaction rotor.

Subsequently, the reaction rotor (7) rotates and transfers the reaction rack (1) to the measuring device θ. This transfer is also controlled by the control device (8)
With the signal from the reaction rack (1) in the same manner as described above,
is transferred. Measurement is carried out using a one-channel vertical photometry method. When the reaction rack (1) passes through the photometry section, each reaction hole r
4) The absorbance of... is sequentially measured one by one.

In order to measure with vertical photometry 78, the transfer belt of the measuring device has a structure that allows the leading vehicle to make a 1i'1 vortex.

? The reaction rack (1) on which the l'111 determination has been completed continues to move, passes through the measurement: 12 apparatus 0 wharf, and is discharged to the outside.

The measured sodium chloride luminosity value is converted into a corresponding concentration value, etc. based on a calibration curve for each item created using a standard solution, etc., and is displayed on the display production device C11.

In the following, the principle of making the present invention kindly follows the measurement process of one reaction rack, but in reality, multiple reaction racks (1) are used, and in some cases, two or more constant items are used. Sometimes it's more than that. Accordingly, "C1" The processing timing of each rack is arranged in chronological order, and the reaction racks that require cleaning, reagent dispensing, or measurement are processed by the corresponding processing equipment, and then sequentially. The analysis process for each rack progresses in such a way that the reaction rack that requires processing is called up and the process is carried out.In order for this analysis process to proceed without any trouble, two or more processes must be performed in a given process step. It is necessary to ensure that two reaction racks are not served at the same time. Therefore, in this example, when a certain reaction rack (1) is undergoing cleaning, reagent dispensing, or measurement, the other reaction racks will not be able to perform any of these processes 1 to 11 at the same time. Control layer f'? (8) is adopted. In other words, when registering a new reaction rack, the reaction section issues an OK to accept instruction because all reaction racks are batch-processed and the timing for cleaning, reagent dispensing, and measurement is determined at the same time. , regardless of the content of the process, it is possible to organize the process timing in chronological order and select a start time such that multiple reaction racks are not involved in one process timing.

Although the following describes one preferred embodiment of the method and apparatus of the present invention, many variations thereof are possible.

First, the n11 example is 21 of the Dupu method and the Sandinch method.
Regarding combination, it can also be applied to bead method and competition method. 1 [1 When using the Sibes method, insert beads and Ql+
A device for transferring the reaction mixture to (((), etc. is required. And, like the cleaning device 01, etc., each of these devices may be appropriately placed around the reaction rotor [7).

On the other hand, if it is legal, you can use it as is, but West Ii
When dispensing the %1 test standard at the same time as the sample, a nozzle (lla) for the drunken smoke test standard 5]rL is installed in the sample dilution/dispensing device Qff1).

Next, in the iQ example, each reaction rack (1) is controlled so that when one reaction rack is undergoing any treatment, the other reaction racks are in the reaction zone. Although the same treatment steps do not overlap, other treatments may be performed in parallel, and the initial reaction start time may be controlled. (3t'y of eyelids, 9.!!! It is necessary to make sure that the reaction rack (1) is completely outside the reaction rotor so that the reaction rotor (7) can rotate during the process. Furthermore, the reaction racks (1) do not have to be returned to their original positions along the paths through which they have undergone each treatment.However, in this case, it is necessary to always remember the position of each reaction rack (1).

In addition, the sample dilution dispensing device (5) and the reagent dispensing device 00 are arranged in parallel with the respective dispensing nozzles, and after each dispensing process, the liquid in each reaction hole (4) is injected into one cup. An endless belt C14) is provided with an air blowing nozzle for stirring, or the reaction rack puller groove is reversibly rotated as shown in Figure 5.
and a pusher (a compact one made of a cocoon, or a separate processing section for purification and dispensing of various reagents at each stage, rather than a common one, as in the 1iC technology), or in the measuring section. Measure from the side instead of vertically,
In addition to transmitted light, methods such as scattered light and fluorescence analysis may be used, or the reaction solution may be separately drawn into a flow cell or the like for optical measurement. Furthermore, since the reaction time lasts for a long time, the temperature of the reaction section may be adjusted to, for example, 37°C, or one sample may be measured using two reaction holes (4), and the horizontal value of both may be used to measure the sample. For example, if the photometric window (1a) of the reaction rack (1) is used as the measuring section of FIG. 6(a), (
The accuracy of the measured value can be improved by using a 7-piece structure that does not rub against the 1"iI!']11 as shown in 1)).

Next, FIGS. 7 and 8 show examples of different reaction tables.

First, the one in Figure 7 is a plate-shaped one in which the reaction table (7) moves in the -axis direction, and the sample is fed from the sample dilution dispenser R (5) while timing the reaction table (7) into the groove on the top surface (as shown). The reaction rack (1) is sequentially provided to each processing device t+t>, (+3 C10) by moving the reaction table (7) in the direction of the arrow. (1 (be111), (move between i and reaction rack (
Analysis of 1) IJJ1/IJJ1. In this case, it is preferable to allow movement in two axial directions without interfering with the movement.

1': In each embodiment, each processing device (1+), (1
0, (6) are arranged around 1.'d of the reaction table (71, (7)), but even if each of these 1 Uraoka is placed tFt above the reaction table, the Figure 9 shows one example, in which a plate-shaped reaction table (71) is made movable in two axes, and any reaction rack (1) at the timing when processing is required is used. is placed below the device that performs the processing, each processing F IQ rl
+i , (IT) , (2) rack movement mh machine <M
becomes unnecessary. It goes without saying that this method can also be applied to the bead method.

Furthermore, in reference to each of the above methods, the reaction table @8 was explained as a combination of multiple units (for example, a reaction rack).
It is also possible to treat individual reaction vessels one by one. However, in this case, especially in the case where each processing device is arranged around a reaction table, it is necessary to use an In-built structure so that each reaction vessel can be easily taken out. Furthermore, although each processing device is fixed at the 11 position on each side, it is also possible to move these devices.

As detailed above, the method of the present invention is an ode to enzyme immunoassay,
The fils of multiple containers or multiple containers (1:) are sequentially received and pooled in the reaction zone, and any container or set of containers at the required timing is used to start the primary reaction. m "It can be used for each treatment process regardless of the order. Therefore, 9/kidney 01!!)
Even if there is a blank space that does not apply, the blank space can be used effectively, and the processing speed is greatly improved. Also, is it possible to set multiple items at the same time? 'fl can be carried out in real time, so efficient use of the apparatus can be achieved, and there is no need to wait for the samples t( to be collected), making it possible to carry out slow measurements.

In addition, with the device of the present invention, there is no restriction on the feeding speed of the sample other than waiting for a processing timing, so the labor and time required for feeding are greatly saved. Simply set the racks and containers, and register the item name and number of samples in the device.
A rapid t11] determination can be carried out completely automatically and continuously. In addition, the operator is freed from the hassle of adjusting the device every time the measurement item changes, and there are fewer manual processes, reducing operational errors.
% l This greatly reduces the mental and physical burden on the author and greatly improves accuracy.

[Brief explanation of drawings]

Fig. 11 is a block diagram of the present invention), Fig. 2 is a schematic oblique view of the main parts, Fig. 3 is a perspective view of an example of a reaction rack, and Fig. 4 is a reaction rack. Move trQ
fP? FIG. 5 is a perspective view showing one example of the reaction rack moving machine t1°, and FIG.
) and (b) are 1? of different reaction racks, respectively. :? The top view and FIGS. 7 to 9 are schematic diagrams showing other embodiments. 1... Reaction rack 4... Reaction hole 5... Sample dilution/dispensing device 6... Operation section 7... Reaction rotor 8... Control device fR10... Device 1
1... Reagent dispensing disease Wlt 12... 1jjll determination device 13... Display recording device Patent applicant L1' Kyoto Daiichi Kagaku No. 7, Figure 8

Claims (1)

[Claims] 1. In enzyme immunoassay, a plurality of 6 containers or multiple T! An automatic enzyme immunoassay method characterized in that sets of containers I are sequentially received in a reaction zone and kept in a pool 17.
62. The automatic enzyme immunoassay method according to claim 1, wherein the set of volume H) or volume H) or volume H) 13 is housed in the reaction zone except when used for each treatment and measurement. 3. The automated enzyme immunoassay method according to claim 2, wherein the container or set of containers is returned to its original position in the reaction pool after each treatment. - The NY system according to claim 1, wherein the four containers or a set of containers are placed in the normal position M of the reaction zone until the end of the measurement, and are moved for each reaction zone and subjected to each treatment and measurement step.
Automatic elementary immune measurement method. When one of the containers or sets of containers is being subjected to any treatment or measurement process, the timing of the start of the primary reaction is controlled so that the container or set of containers is waiting in the reaction zone.
The automatic enzyme immunoassay method according to claim 1, 2, 3, or 4, wherein the enzyme immunoassay method is a method for automated enzyme immunoassay. While one of the containers or sets of containers is being processed in any treatment or measurement step, the other container or set of containers is introduced into another treatment or measurement step; Claim 1, which controls the timing of reaction initiation;
The automatic enzyme immunoassay method according to item 2 or 3. 5. The automatic enzyme immunoassay method according to claim 1, 2, 3, or 4, wherein the reaction zone is commonly used for each step of the reaction. 8. The automatic enzyme immunoassay method according to claim 1, 2, 3, or 4, wherein the reaction zone is divided into a plurality of zones so as to correspond to each stage of reaction. 9. The enzyme according to claim 1, 2, 3, or 4, wherein the washing and/or dispensing steps of various reagents at each stage are performed in a common processing section. Automatic immunoassay method. 10 After contacting and reacting the test solution with a substance that specifically reacts with a specific component in the test solution in a container, cleaning the carrier at each stage and dispensing various reagents into the container. In an apparatus for optically measuring a reaction and a reaction solution, on the surface of a movable or rotatable reaction table which stores a plurality of containers or sets of containers after the start of the next reaction, one set of containers from the reaction table is placed. Alternatively, a cleaning device, a dispensing device, and a measuring device are installed to sequentially receive and process sets of containers, and at least a primary system is installed to prevent multiple containers or sets of containers from being involved in the same processing process. An automatic enzyme immunoassay device 011 characterized in that it is equipped with a control device that selects the reaction start time and controls the operation of each device.Claim 10, which is characterized in that it is equipped with a device for transferring a carrier or liquid in a container. The automatic enzyme immunoassay device described in Section 1. 12 Claim 1 in which the reaction table is rotor-shaped
The automatic enzyme immunoassay device according to item 0. 13. The automatic enzyme immunoassay device according to claim 10, wherein the reaction table moves in one or two axes. 14. The automatic enzyme immunoassay 'li R according to claim 10, wherein the reaction table is a plurality of plate-shaped plates each of which moves in biaxial directions. 15 After contacting and reacting the test solution with a substance that specifically reacts with a specific component in the test solution in a container, washing the carrier at each stage, dispensing various reagents into the container, and In an apparatus for optically measuring a reaction and a reaction solution, a cleaning device, a dispensing device, and a measuring device are installed above a movable or rotatable reaction table that accommodates a plurality of containers or sets of containers after the next reaction has started. A control device is provided for selecting the start time of the primary reaction and controlling the operation of each device so that all processing operations for a plurality of containers or a set of containers in the reaction table A are not duplicated. An automatic enzyme immunoassay device characterized by: 16. The automatic enzyme immunoassay device according to claim 15, comprising a carrier in a container or a liquid transfer device.