JPH02228564A - Automatic immunity measuring apparatus - Google Patents

Abstract

PURPOSE:To control an amount and a timing of a buffer liquid with an ON-OFF operation only of an solenoid valve by connecting a heater for heating the buffer liquid and a solenoid valve for controlling the supply thereof at each position of supplying the buffer liquid of a reaction turntable to allow a common use of a pump. CONSTITUTION:A reaction detection container 8 is set on a reaction turntable having a plurality of positions to measure an amount of an antigen or an antibody of a sample quantitatively by an immunoreaction between an antigen or an antibody thereof. Then, a resistance tube 5, a heater 7 for heating the buffer liquid and solenoid valves 3 and 5 for controlling the supply thereof are connected at each position of supplying the buffer liquid of the reaction turntable from a pressurizing tank 2 to the position. Thus, a flow rate can be controlled with the resistance tube 5 without controlling flow systems with individual pumps thereby enabling the supplying of the buffer liquid heated up to a proper temperature with the heater 7 at a free timing by an ON-OFF operation of the solenoid valves 3 and 5.

Description

DETAILED DESCRIPTION OF THE INVENTION [Field of Industrial Application] The present invention relates to an automatic immunoassay device that automatically measures components contained in a sample such as serum or urine through an immune reaction using a solid phase reagent.

[Conventional technology]

Enzyme immunoassay is a method that uses enzyme activity as a marker to determine the extent of antigen-antibody reaction, and then quantifies the amount of antigen or antibody. Furthermore, there are various methods such as the Sand-Deutsch method in which an enzyme-labeled antigen (or antibody) is reacted, the competitive reaction method in which an enzyme-labeled antigen (or antibody) and an unlabeled antigen (or antibody) are competed to obtain the unlabeled antigen machine, and various other methods. There is a way.

FIG. 5 is a diagram for explaining the principle of measurement by the Sand-Deutsch method, and FIG. 6 is a diagram for explaining the principle of measurement by the competitive method, in which 61 is a solid phase carrier, 62 is an antibody, 63 is an antigen to be measured, 64 is a labeling substance, 65 is an antibody, 66 is a substrate, 67
indicates the product.

As shown in FIG. 5, in the Sand-Deutsch method, first, an antibody 62 is immobilized on the surface of a solid phase carrier 61 by physical adsorption or chemical reaction, and an antigen to be measured 63 is attached to this. Add samples containing.

(2) As a result, an antigen-antibody reaction (first reaction) occurs and the antigen to be measured 63 binds to the immobilized antibody 62, so that the extraneous components in the sample are then discarded by washing.

(2) Next, an enzyme-labeled antibody 65 to which an enzyme is bound as a labeling substance 64 is added.

(2) As a result, an antigen-antibody reaction (second reaction) occurs, and the enzyme-labeled antibody 65 binds to the antigen 63 bound to the immobilized antibody 62. In this case, since the enzyme-labeled antibody 65 is added in excess, a bound portion (b) formed by binding the antigen 63 and the antibody 65.

und SB) and the free part (fre
e, F) can be done.

■ Therefore, by washing, free products (f
F) Discard the excess enzyme-labeled antibody.

That is, B/F separation is performed.

■ Next, perform an enzymatic reaction. Since the enzyme activity at this time is determined by the amount of enzyme-labeled antibody in the bound portion, the amount of enzyme-labeled antibody represents the amount of antigen bound to the immobilized antibody, that is, the amount of antigen to be measured.

In the competitive reaction method, as shown in FIG. 6, (1) An antibody is immobilized and wiped, and an antigen to be measured and a labeled antigen in which a label is bound to the same antigen as the antigen to be measured are added.

(2) As a result, an antigen-antibody reaction occurs, and the antigen to be measured and the MA recognition antigen bind to the immobilized antibody in proportion to their respective amounts.

(2), (2) Next, B/F separation is performed in the same manner as the Sandersch method, and an enzymatic reaction is performed. Since the enzyme activity at this time is determined by the amount of enzyme-labeled antibody in the bound portion, the amount of the antigen to be measured can be determined from the amount of labeled antigen added using a calibration curve.

All of the above methods are so-called separation methods, and there are also non-separation methods that do not perform B/F separation.

Although the non-separation method is quick in measurement time and easy to operate, it has drawbacks such as low measurement sensitivity. In comparison, separation methods have high measurement sensitivity, but are extremely complex and cumbersome to operate, and are difficult to automate (previously, they were performed manually in most cases).

When the above-mentioned enzyme immunoassay is automated, an antibody or antigen immobilized on a solid phase carrier such as a polystyrene ball or glass beads is used as a solid phase reagent. Since this solid phase reagent is unstable, it is usually kept in a container filled with a storage solution, and when performing enzyme immunoassay, the solid phase reagents are transferred one by one from the container to the reaction detection container. , sample dispensing, labeling reagent dispensing, B/F separation, washing, etc., and then the bound portion is transferred to the detector.

[Problem to be solved by the invention]

In the above-mentioned apparatus, the solid phase reagent is transferred after the surface phase reagent is transferred to the reaction detection container, after the first reaction caused by dispensing the sample, and after the second reaction caused by dispensing the labeled reagent. In order to perform washing and B/F separation, a flow system is provided to supply a washing liquid. Each of these flow systems is equipped with a pump to supply cleaning fluid. This increases the cost of the device. Therefore, if you try to share a pump for each flow system, if all flow systems supply cleaning liquid at the same time, or if some flow systems do not supply cleaning liquid, the flow rate will change and the quantitative accuracy will be affected. The problem arises that it cannot be maintained. In other words, in the past, a pump was required for each flow system in order to maintain the quantitative nature of supplying the cleaning liquid.

The present invention is intended to solve the above-mentioned problems, and aims to provide an automatic immunoassay device that uses a common pump and can set the amount and timing of cleaning liquid only by turning on and off a solenoid valve. It is.

[Means to solve the problem]

To this end, the present invention provides an automatic immunoassay device that quantitatively measures the amount of an antigen or antibody in a sample by setting a reaction detection container on a reaction turntable having multiple positions and performing an immunoreaction with the antigen or antibody in the sample. A pressurized tank containing a buffer solution and a heater that heats the resistor and buffer solution for each buffer supply position of the reaction turntable are controlled between the pressurized tank and the position, and the supply of the buffer solution. It is characterized by having a flow system connected to a solenoid valve.

[Effect]

In the automatic immunoassay device of the present invention, for each buffer solution supplying position of the reaction turntable, there is a heater that heats the resistor and the buffer solution between the pressurized tank and the buffer solution supply position, and an electromagnetic sensor that controls the supply of the buffer solution. Since it is connected to a valve, the flow rate is controlled by the resistance tube without having to control each flow system with a separate pump, and the buffer solution heated to an appropriate temperature by the heater can be controlled at any timing by turning on and off the solenoid valve. can be supplied to each position.

〔Example〕

Examples will be described below with reference to the drawings.

FIG. 1 is a diagram for explaining one embodiment of the automatic immunoassay device according to the present invention, in which 1 is a compressor, 2 is a tank, 3 and 6 are electromagnetic valves, 4 is a buffer solution, 5 is a resistance pipe, 7 is a preheater, and 8 is a reaction detection container.

In FIG. 1, a buffer solution 4 is used as a cleaning solution or a diluent, and a tank 2 is a pressurized tank that contains this buffer solution 4 and is supplied with pressurized air from a compressor 1 through a solenoid valve 3. It is.

For example, assuming that the tube for supplying the buffer solution 4 is 2 mm in diameter, the resistance tube 5 is constructed of a thin tube of about 0.5 mm in diameter, and the flow rate of the buffer solution 4 pressurized in the tank 2 can be adjusted by adjusting the length. This is to set. The solenoid valve 6 controls the supply of the buffer solution 4 by turning on/off, and the preheater 7 heats the buffer solution 4 supplied to the reaction detection container 8 to about 37° C., for example. The tube may be immersed in hot water that is controlled at a constant temperature.

As shown in the figure, if a pressurized tank 2 is shared and a resistance pipe 5, a solenoid valve 6, and a preheater 7 are connected to each flow system that supplies buffer solution 4 from this tank 2, a plurality of Even if the same buffer solution 4 is injected into the positions at the same time or at different timings, the flow rate can be set for each flow system using the resistance tube 5, and the buffer solution 4 can be supplied using the solenoid valve 6. It can be controlled and quantitativeity can be maintained. In addition, when a reaction detection container as described below is used, the compressor 1 can also be used to supply pressurized air in the cleaning process. I use compressed air, but you can also use it for this as well. Therefore, compared to conventional devices having pumps for each flow system, it is possible to significantly reduce costs.

Next, an automatic immunoassay device to which the present invention is applied will be explained.

Fig. 2 is a diagram showing one embodiment of a cartridge used in the automatic immunoassay device according to the present invention, Fig. 3 is a diagram showing one embodiment of the entire flow system, and Fig. 4 is a diagram showing one embodiment of the layout. FIG.

In FIG. 1, 11 is a cartridge body, 12 is a filter, 13 is a solid phase reagent, 14 is a discharge hole, 15 is an opening, 16 is an aluminum cap, 17 is an orifice, and 18 is a tip.

The cartridge shown in FIG. 2 is a disposable reaction detection container, and has a cylindrical cartridge body 11 containing a solid phase reagent 1.
3, and a filter 12 is provided below it.Furthermore, a narrow discharge hole 14 is provided halfway under the filter 12, and an opening 15 at the upper end is provided with an aluminum cap 16.
It is blocked by. The solid phase reagent 13 is made by immobilizing antibodies on the surface of similar particles with a diameter of about several tens of μm. Since the antibodies and antigens in the solid phase reagent 13 are proteins and are easily decomposed, they should not be treated with preservatives or at a certain pH. It is immersed in a preservation solution such as M fresh solution to preserve it.

Further, there is an orimeter 17 cut in the part where the ejection hole 14 is provided, and by bending the tip part 18 at the orimeter 17, the cartridge can be easily removed from the cartridge main body 11, and the ejection hole 14 can be passed through. be able to. The discharge hole 14 is formed with an extremely small diameter, and
Since the filter 12 is arranged, even when the distal end 18 is removed from the cartridge body 11 when using the car IJ, dispensed sumbu, reagents, washing water, etc. can still flow through the discharge hole 14. The gas is not easily discharged from the air, and is discharged by supplying pressurized air from the opening 15 at the upper end or by suctioning from the discharge hole 14.

Therefore, the cartridge has an aluminum cap 1 at the top end.
6, the solid phase reagent 13 is stored on the filter 12 with its lower end completely sealed by the tip 18, and is stored in a cartridge turntable having an apparatus configuration as described below. When this cartridge is transferred from the cartridge turntable to the reaction turntable by the arm mechanism, the tip 18 is removed from the dust, and the storage solution is discharged at the next position on the reaction turntable for cleaning.

Next, the flow system of the automatic immunoassay device will be explained.

FIG. 3 is an overall flow diagram, in which 21 is a reaction turntable, 22 is a cartridge turntable, 23 is a reagent turntable, 24 is a sample turntable, 25 is a sample cup, 26 is a disposable chip, 27 is an arm mechanism, 28 is a detector, 29 is a dust, 31 is a drain tank, 32 is a compressor, 33 is an in-out switching valve, 34-37. 51 and 53 are pumps, 38-4
1 is the tank, 42 is the 3-way joint, 43 is the resistance tube, 4
4 is a preheater, 45, 52 and 54 are valves, and 46 is a mixer.

As shown in FIG. 3, the flow system is connected to the reaction turntable 21 having 29 positions, with the position (2) as the starting point, and flow systems for dispensing samples and reagents, washing, etc.

At the base position (3), the cartridge is first set on the reaction turntable, and the reaction turntable is alternately rotated through two predetermined positions. The cartridge after the reaction is transferred to the detector 28.

First, each flow system connected to the reaction turntable 21 will be explained in FIG.

The drain tank 31 is for storing the storage solution and cleaning solution discarded from each cartridge of the reaction turntable 21, and is connected to a drain path provided in an annular manner below each position of the reaction turntable 21. The compressor 32 is used for discarding storage solution, cleaning immediately after that, and B/
It supplies pressurized air for cleaning during F separation, disposal of samples and reagents remaining at the tip of the disposable chip, and cleaning.

The tank 38 is for storing the luminescent auxiliary reagent, and the tanks 39 and 40 are for storing the luminescent reagent, respectively.The in-out switching valve 33 and the pumps 34 to 37 are for storing the luminescent auxiliary reagent, the diluent, and the luminescent reagent. It is something. A buffer solution stored in the tank 41 is used as the diluting solution and the washing solution. This buffer is, for example, a mixture of surfactants, sugars, etc. that promote immune reactions. The tank 41 has a sealed structure and is configured to supply pressurized air from the compressor 32 and apply pressure to feed the cleaning liquid. By controlling the temperature and flow rate to a stable predetermined temperature and flow rate, the reaction is facilitated and stabilized. Similarly, for the luminescent reagent, a heater may be added to the mixer 46 to stabilize the temperature during the luminescent reaction. In addition, when injecting the luminescent reagent, since the mixed material remains in the tube beyond the three-way joint 42, it is necessary to spit it out immediately before injecting the luminescent reagent.

For example, in the pump suction step with the in-out switching valve 33 in the state shown, the pump 34 suctions the luminescent auxiliary reagent from the tank 38, the pump 35 suctions the diluent (water) from the tank 41, and the pump 36.
37 simultaneously aspirates luminescent reagent from tanks 39 and 40. Then, when the in-out switching valve 33 switches (the upper half shifts to the right) and enters the pump discharge process, the luminescent auxiliary reagent and the diluent are heated to a predetermined temperature through the preheater 44 and are heated to the cartridges in positions ■ and ■, respectively. is injected into. Further, the luminescent reagent is mixed through a three-way joint 42 and a mixer 46,
is injected into the cartridge of the detector 28.

In the cleaning process, the valve 45 is selectively opened and closed until t! !
The new buffer solution is injected from the tank 41 through the resistance tube 43, preheater 44, and valve 45 into the cartridges at the respective positions ①, ②, and ②. Then, the air valves are selectively opened and closed, and pressurized air is supplied from the compressor 32 to the cartridges at the respective positions (1), (2), and (2) through the air valves. In normal cleaning, injection of a cleaning solution (buffer solution) and disposal using pressurized air are performed four times.

In position ■, the sample dispensing flow system and the labeled antibody reagent dispensing flow system are connected so that the Sand-Deutsch method and the competitive reaction method can be applied, but these are each connected to a dedicated disposable from the sample cup or reagent bottle. It is inhaled and dispensed using a tip.

In this case, sample or reagent remains on the tip, so
A pressurized air flow system is connected to blow them out with pressurized air. In the sample dispensing system, this is the flow system of the valve 52, and the tip of the disposable chip is inserted into the sample cup of the sample turntable 24, and the sample is sucked by the sampling pump 51 and dispensed into the cartridge at position ■. You can switch to this system later. Similarly, when dispensing a reagent, insert the tip of the disposable tip into the reagent bottle of the reagent turntable 23, suck it in and dispense it with the reagent pump 53, and then connect it to the pressurized air flow system with the valve 54. Can be switched. Also,
If the internal pressure increases, the amount of suction becomes unstable, so a valve is also provided to release the air to the atmosphere.

Next, the rotation of the position of the reaction turntable 21 will be explained.

At position (2), the arm mechanism 27 operates to transport a new cartridge from the cartridge turntable 22, remove the tip, and set it.

Tip shown in Figure 2 when setting a new cartridge in position ■! Even if 28 is removed from the cartridge, the storage solution inside will not be discarded, so pressurized air is sent from the upper opening of the cartridge in position ① to discard the storage solution inside the IJ cartridge.

At the next position (3), the cleaning valve is set at the upper end opening of the cartridge, and cleaning is performed by repeatedly sending cleaning liquid and pressurized air alternately, for example, four times.

Next, at position ■, add the diluent. This is to make it easier to cause an immune reaction, since when blood, serum, urine, etc. are directly injected, various components may interfere with the immune reaction.

Then, at position ■, the sample is aspirated from the sampling cup with a disposable tip and dispensed into the cartridge.

After that, the immune reaction (first reaction) is promoted by applying vibration and stirring each time it rotates one position at a time, and at position 0, water is supplied and water is drained using pressurized air, which is repeated four times for cleaning. Perform the B/F separation as described.

Here, we will explain in detail the operation after B/F separation.
First, rotate forward from position 0 to 20 positions, stop at position ■, and add diluent ($
Inject 1 equilibration solution) and rotate it further 10 positions in the forward direction until it reaches position ■, which is 1 position ahead of the previous one. After stirring by vibration, the tube is similarly rotated 20 positions in the forward direction, and once stopped at position (3), the labeled antibody is dispensed. The diluent has the effect of stabilizing the reaction temperature by preheating, facilitating and promoting the immune reaction, and enhances the stirring effect when the next labeling reagent is dispensed. This is the operation in the case of the Sanderutsch method.

That is, by alternately performing an operation of rotating the pitch of 10 positions and an operation of rotating the pitch of 20 positions, the position is advanced one position at a time from the previous position. By doing this, it is possible to adopt an apparatus configuration in which the labeling reagent is dispensed at the sample dispensing position ① even in the Sand-Deutsch method. As a result, by controlling the rotation operation so that the sample and labeled antigen are simultaneously dispensed at position ■, it is possible to perform immunoassays using the same flow system even in the competitive reaction method. Similarly to the reaction, the immunoreaction (second reaction) is promoted by applying vibration and stirring each time it rotates one position at a time, and B/F separation by washing is performed again at position (3).

Then, a luminescence auxiliary reagent for intensifying luminescence is added at position (■), and the cartridge is transferred to the detector 28 at the initial position (■). The detector 28 measures the amount of luminescence immediately after adding the luminescence reagent to the cartridge. The luminescent reagent varies depending on the labeling substance, but for example, acridinium
um), a mixture of hydrogen peroxide and an alkali, and Luminol, a mixture of hydrogen peroxide and F.
A mixed solution of e-ions is used, but since these react in a short time, they are injected simultaneously from each bottle through a three-way joint 42 and a mixer 46.

In the above operation, for example, it stays at position ■ for 12 seconds, rotates in the forward direction at a pitch of 20 positions, stays at position 0 for 12 seconds, then rotates at a pitch of 10 positions to position ■, and then rotates for 24 seconds. Then, the cartridge is moved from position ■ to position ■, ..., one position at a time, and at position ■, the cartridge is first transferred to the detector 28 in 12 seconds, and then a new cartridge is brought from the cartridge turntable 22 and set. I will do it. In this case, the first reaction takes about 3 minutes, the second reaction takes about 5 minutes, and the immune reaction measurement for one sample can be performed in about 10 minutes in total, and 150
It is possible to achieve a measurement speed on the order of 100 ms/hr.

In addition, when measuring multiple items with one sample, in position
2, the same sample is dispensed into those cartridges, and a reagent corresponding to the measurement item is further dispensed. Then, the amount of light emitted is measured by the detector 28, and the process ends. If measurement is required again, the remaining disposable tip 26 is used to dispense the sample. For this purpose, the sample turntable 24 stores two disposable chips for each sample. Re-measurement is required not only when the measured value shows an abnormal value that significantly deviates from the preset range, but also when a judgment value is given for each measurement item in advance as a secondary screening, and the next measurement item is determined based on the judgment result. may be set. For example, in a serum hepatitis test, first a test for HBS antigen is performed, and if the result is positive, a test for HBE or HBS antigen is performed. In addition, the reason why we use disposable tips made of Japanese cypress is that the range is very wide in the case of immunoassays, and complete cleaning cannot be done with conventional pipettes.

FIG. 4 shows an example of the layout of an automatic immunoassay device.

In FIG. 4, 55 and 56 are arm mechanisms, and 57 is a control processing unit. Cartridge turntable 22
is a car made of Japanese cypress as shown in Figure 2) IJji (
This is a table that stores and rotates a reaction detection container (reaction detection container), and shows an example in which it is made up of 10 removable cassettes and each cassette can store 30 cartridges. According to this, since cartridges of the same immobilized antibody are stored in each section, cartridges for 10 items can be prepared. Reagent turntable 2
3 is a rotating table that stores reagent bottles for labeled antibodies and disposable chips for dispensing the labeled antibodies, and shows an example in which reagent bottles for 10 types, that is, 10 items can be stored. The sample turntable 24 is a rotating table that stores a sample cup 25 containing a sample and a disposable chip 26 for dispensing the sample, and has two disposable chips 26 inside thereof corresponding to each sample cup 25. An example is shown in which it can be stored. The reaction turntable 21 is used to perform dispensing of samples, addition of labeled antibodies, stirring/reaction by vibration, washing, etc., as described above, while the cartridges of the cartridge turntable 22 are set and rotated by l positions. be. The arm mechanisms 27, 55, and 56 are mechanisms for inserting and removing cartridges and dispensing reagents and samples between the reaction turntable 21, cartridge turntable 22, reagent turntable 23, and sample turntable 24. , circles a and b%C show the respective trajectories.

Further, the detector 28 is for injecting a luminescent reagent into the cartridge after reaction and detecting the amount of luminescence, and the dust 29 is for discarding the cartridge after the amount of luminescence has been detected.

Note that the present invention is not limited to the above embodiments, and various modifications are possible. For example, in the above embodiment, each flow system is drawn from the tank separately, but if the flow resistance in the flow system pipes is sufficiently small,
It may be configured such that one pipe is drawn out from the tank and then branched off. Further, although a thin resistance tube is used for setting the flow rate, a variable valve such as a needle valve may also be used. In terms of the overall device configuration, automatic luminescence immunoassay using the Sand-Germansch method has been explained, but the device configuration shown in Figures 3 and 4 also allows automatic luminescence immunoassay using the competitive reaction method, and can be used for other immunoassays instead of luminescent immunoassay. Of course, it can also be applied to measurement methods such as enzyme immunoassay. Furthermore, although the cleaning liquid and pressurized air are alternately supplied four times, it goes without saying that this number of times can be changed as appropriate.

〔Effect of the invention〕

As is clear from the above description, according to the present invention, the flow rate is controlled by the resistance section without the need to control each flow system with an individual pump, and the t1 equilibrium liquid heated to an appropriate temperature by the heater is electromagnetically heated. Supply can be made to each position at any time by turning the valve on/off. Moreover, even if the same buffer solution is injected into multiple positions at the same time or at different timings, the flow rate can be set for each flow system using a resistance tube, and the buffer solution can be supplied using a solenoid valve. can be controlled and quantitative properties can be maintained.

Therefore, compared to conventional devices having pumps for each flow system, costs can be significantly reduced and the configuration of the flow system can be simplified.

[Brief explanation of the drawing]

FIG. 1 is a diagram for explaining one embodiment of the automatic immunoassay device according to the present invention, FIG. 2 is a diagram showing one embodiment of a cartridge used in the automatic immunoassay device according to the present invention, and FIG.
The figure is an overall flow diagram, Figure 4 is a diagram showing an example of the overall layout, Figure 5 is a diagram to explain the measurement principle by the Sandersch method, and Figure 6 is a diagram to explain the measurement principle by the competitive method. It is a diagram. 1...Compressor, 2...Tank, 3 and 6...
・Solenoid valve, 4...Buffer solution, 5...Resistance tube, 7...
Preheater, 8...reaction detection container. Applicant: JEOL Co., Ltd. Agent: Patent attorney Ryukichi Abe (5 others) Figure 4 (b)

Claims (2)

[Claims] (1) An automatic immunoassay device that quantitatively measures the amount of an antigen or antibody in a sample through an immune reaction with the antigen or antibody in the sample by setting a reaction detection container on a reaction turntable with multiple positions, and in which a buffer solution is used. A pressurized tank accommodated therein, and a heater for heating the resistor and the buffer solution for each position of the reaction turntable for supplying the buffer solution between the pressurized tank and the position, and a solenoid valve for controlling the supply of the buffer solution. An automatic immunoassay device characterized by having a flow system connected to. (2) The automatic immunoassay device according to claim 1, further comprising a compressor, and the pressurized air obtained by the compressor is used for draining liquid in the reaction detection container and pressurizing the pressurized tank.