JP2006017736A - Member for collecting microorganisms and immunity measuring apparatus - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide an immunity measuring apparatus capable of automatically carrying out high precision and rapid immunity measurements. <P>SOLUTION: The immunity measuring apparatus comprises a pretreatment unit which is composed of a filtering section 101 for a sample liquid including a microorganism being an immunity measurement object, an condensed liquid producing section 102, a collection vessel 103 for collecting the microorganism that is the immunity measurement object into a reaction liquid, and an ultrasonic crushing section 105 for crushing the collected microorganism by using ultrasonic waves and for diffusing antigens in the microorganism in the reaction liquid; an immunoreaction unit 111 which carries out antigen-antibody reactions of the antigens and antibodies in the reaction liquid; a luminous reaction unit 115 which causes a luminescence reagent to react with an antigen-antibody complex generated by the immunity reaction; a moving unit for equipments being used for the immunity measurement; and a control section which controls respective units and automates the immunity measurement of the microorganism. <P>COPYRIGHT: (C)2006,JPO&NCIPI

Description

  The present invention relates to an immunoassay device, and more particularly to an immunoassay device in which immunoassay is automated.

  Immunoassays, which are quantitative methods for measuring the amount of antigen or antibody, can be classified into various immunoassays based on the measurement target, the measurement method, the reaction format, etc., but the principle is based on the antibody-antigen reaction.

  Therefore, non-radioimmunoassay methods and non-labeled immunoassay methods that do not use radioisotopes are not so different in the operation of each immunoassay method except that the final measurement system is different.

  Such immunoassay is required in various fields. The following is an example of applying immunoassay to the measurement of coliforms.

  In Japan, it is stipulated that the number of coliforms in effluent treated at a sewage treatment plant must be 3000 or less (ml) (Sewerage Law).

  Therefore, in each sewage treatment plant, the treated water is finally brought into contact with chlorine and sterilized, and then discharged. At present, the quantitative tests for the number of coliforms include a plate culture method using a desoxycholate medium, a most probable number method (MPN method), and a membrane filter method. Among these, the plate culture method using desoxycholate medium is used in the verification method related to the drainage standard established by the Ministerial Ordinance on the Water Quality Test Method and the Director-General of the Environment Agency, and the most probable number method is used in the environmental standard related to water pollution. (MPN Law) is defined as an official law.

  The plate culture method using desoxycholate medium (hereinafter referred to as “deso method”) uses a commercially available desoxycholate medium to obtain a typical colony unique to coliforms that is generated when plate culture is performed. This is a method for obtaining the number of coliform bacteria in the measurement target. The operation procedure is shown below.

    (1) Enter the sample name and dilution factor on two or more petri dishes. In this case, several diluted samples are cultured so that the number of colonies in one petri dish after culturing falls within the range of 30 to 300.

    (2) Using a measuring pipette, take 1 (ml) of the sample correctly and aseptically place it in each petri dish.

    (3) Aseptically add 10 ml of desoxycholate medium kept at about 45 ° C. to each dish.

    (4) The sample and the medium are thoroughly mixed while rotating the back and forth and right and left carefully so that the agar does not harden, and after thoroughly dispersing on the whole surface of the petri dish, the sample is left on a horizontal plate and allowed to cool.

    (5) Further, add 5-10 (ml) of medium and layer.

    (6) When the medium is completely solidified, invert the petri dish and incubate at 35-37 ° C. for 18-20 hours.

    (7) After culturing, the regular colonies (circular or rice granular) exhibiting red to deep red are counted, and the number of coliforms in sample 1 (ml) is determined. In addition, suspicious colonies are inoculated into a BGLB fermentation tube with a platinum loop and then cultured at 35-37 ° C. for 48 hours.

  Currently, it is desired to measure coliform bacteria in a short time and to save labor by automating the measurement, but the above-mentioned deso method is not only complicated and requires skill, but also until a measurement result is obtained. Since it takes 18 hours or more, it has not been possible to use the measurement result as a chlorine injection index. Therefore, the actual situation is that proper chlorine injection amount control is not performed at present.

  Therefore, it is desired to automate the measurement to save labor and shorten the measurement time. However, the above measurement method is difficult to automate and requires culture for about 18 to 20 hours. It is impossible to reduce the time any further.

  Therefore, it is desired to apply immunoassay to automate the measurement of coliform bacteria and shorten the measurement time.

  Thus, in order to apply immunoassay not only to the measurement of coliform bacteria but also to various fields, there is a need for an immunoassay apparatus capable of performing measurement automatically in a short time.

  However, conventional immunoassay methods require a long time for measurement and are difficult to automate.

  Therefore, the applicant of the present application previously described the test tube (collection container) to which the antibody was bound by the antibody-binding solid phase preparation method in Japanese Patent Application No. 4-112478, Japanese Patent Application No. 4-112479, etc. An immunoassay method was proposed as a reaction container.

  The reaction container to which the antibody is bound is prepared by preparing an antibody-binding solid phase in which an antibody that specifically reacts only with the antigen to be measured in a solution is previously bound to a polystyrene test tube and binding the antibody. can get. The outline is shown below.

  FIG. 41 shows a procedure for measuring a trace amount of antigen in a solution by a luminescence immunoassay using a marker (particularly enzyme) -labeled antibody, a luminescent substance and an antibody-bound test tube. In addition, the test tube to which the antibody is bound is prepared by preparing an antibody-binding solid phase in which an antibody that specifically reacts only with the antigen to be measured in a solution is previously bound to a polystyrene test tube and binding the antibody. can get.

    (1) A predetermined amount of a solution containing the antigen to be measured is dispensed into a test tube.

    (2) Incubate at a constant temperature for a fixed temperature (incubate at a constant temperature).

    (3) Dispense a certain amount of enzyme-labeled antibody into (2) manually using an instrument.

    (4) Incubate for a certain time at a certain temperature.

    (5) Wash the test tube.

    (6) A predetermined amount of a luminescent reagent solution that reacts with an enzyme-labeled antibody and chemiluminescences is dispensed into (5).

    (7) The amount of chemiluminescence in (6) is detected by a sensor.

  In the operation (2), the antigen is bound to the antibody-bound solid phase by the antibody-antigen reaction, and in the operation (4), a sandwich structure complex of antibody-antigen-enzyme labeled antibody is formed.

  In the operation (7), the amount of chemiluminescence emitted by the reaction between the complex of the sandwich structure of antibody-antigen-enzyme labeled antibody and the luminescent reagent is quantified. The concentration of the antibody-antigen-enzyme labeled antibody (that is, the antigen concentration in the solution) can be quantified based on the luminescence amount and the calibration relationship between the enzyme-labeled antibody concentration and the luminescence amount determined in advance.

  Thus, when performing immunoassay, quantification of a solution becomes important, and it is preferable to use a container that is highly accurate and easy to handle as a container used for measurement.

  As containers for sample solutions, beakers, Erlenmeyer flasks, or similar containers are commercially available, and since they are easily available, these containers are used for devices that automate some or all of immunoassays. Yes.

  In the immunoassay, microorganisms are quantified using an antigen-antibody reaction. At that time, it is necessary to concentrate the sample solution to a concentration sufficient for immunoassay. An explanatory view of a conventional concentration process is shown in FIG.

  As shown in this figure, the sample solution to be concentrated is first filtered under reduced pressure through a filter 4 set in a concentrator, and organisms are obtained by filtration (mainly microorganisms, hereinafter, organisms to be filtered are collectively referred to as microorganisms). The filter 4 that has captured is put into a beaker 5. Further, the buffer solution and the beads 2 are added and shaken by the shaker 6, and the microorganisms attached to the filter 5 are washed away and dispersed and collected in the buffer solution.

  Thereafter, the buffer solution from which the microorganisms have been collected is transferred to another container to obtain a concentrated solution. At this time, the concentration ratio is determined by the ratio of the original sample solution and the buffer solution. Usually, the ratio of the original sample solution and the buffer solution is used as it is as the concentration factor.

  According to such an immunoassay method, it is possible to measure the coliform group and the like in about 2 hours.

  However, immunoassays that can be performed in a short time as described above must be performed manually. As long as a human operation is performed in this way, accidental errors and systematic errors always occur, so there is a limit to variations in measurement accuracy and improvements in measurement accuracy.

  In particular, the measurement of trace amounts of antigen in a solution by a luminescent immunoassay using a labeled marker (particularly an enzyme), a luminescent substance and an antibody-bound solid phase is a complicated and complicated procedure, and it is difficult to obtain high accuracy. is there.

  Therefore, it is required to automate immunoassay and increase measurement accuracy. At present, some immunoassays can be automated using general-purpose devices, but these devices have the following problems.

  Although it is not difficult for humans to perform filtration operations, it is not possible to meet the demand for automation as it is. This is because these instruments do not have shapes and mechanisms that can be automated. Therefore, a devised shape / mechanism for automation is required.

  In addition, in the concentration step, it is necessary to put in and out a predetermined number of beads into the beaker, but this step is complicated and takes time. In particular, in automating immunoassay using an immunoassay apparatus, this process is expensive.

  Furthermore, when extracting the concentrated solution after shaking the container to collect the microorganisms in the buffer solution, the suction nozzle is operated by a robot, etc., but the tip of the nozzle hits the bead in the beaker and the nozzle is damaged. Or the concentrate may not be removed completely.

  This is because it is uncertain where the beads are located after the shaking operation. To eliminate this, the beads can be removed after shaking, but it is very expensive to mechanize.

  The present invention has been made under the above-described background, and an object thereof is to provide an immunoassay apparatus that eliminates the complexity of manual work as much as possible.

  In order to solve the above-mentioned problem, the invention described in claim 1 is a member for collecting microorganisms that mixes microorganisms captured by a filtration membrane with a liquid, wherein the microorganisms are brought into contact with the filter in which the microorganisms are captured. Provided is a member for collecting microorganisms, characterized in that it has a plurality of bead bodies to be dispersed, a holder section, and a string section provided on each bead body and connecting the holder section and each bead body.

  The invention according to claim 2 is a member for collecting microorganisms that mixes microorganisms captured by a filtration membrane with a liquid, and a dispersion plate that contacts the filter in which the microorganisms are captured and disperses the microorganisms; And a disc portion having a holder portion attached in a direction perpendicular to the dispersion plate, and the dispersion plate has a plurality of holes, and provides a member for collecting microorganisms.

  Further, the invention according to claim 3 is a collection unit for collecting the immunoassay target microorganism in a reaction solution from a sample liquid containing the immunoassay target microorganism, and an antigen in the microorganism by performing ultrasonic crushing of the recovered microorganism. A pretreatment device having an ultrasonic crushing part that diffuses the reaction solution into the reaction solution, an immunoreaction device that causes an antigen-antibody reaction between the antigen and the antibody in the reaction solution, and an antigen-antibody complex produced by the immune reaction A luminescence reaction device that reacts with a luminescent reagent, a device moving device used for immunoassay, and a control unit that controls each device to automate the immunoassay of the microorganism, and the recovery unit includes the A filtration unit that filters the sample liquid and captures the microorganisms in a filtration membrane; and a concentrated solution generation unit that diffuses the microorganisms captured by the filtration membrane into the reaction solution to generate a concentrate, and The ultrasonic crushing part is the concentrated liquid A first recovery container to be injected; and an ultrasonic generator. The immune reaction device includes a second recovery container in which an antibody is fixed to an inner wall thereof to act as a solid phase antibody, and an ultrasonic crushing unit. A nozzle part for sucking a predetermined amount of the reaction solution in which the antigen produced in step 1 is diffused and injecting it into the second recovery container; a labeled antibody injection part for injecting the labeled antibody into the second recovery container; And the luminescence reaction apparatus includes a washing unit for the second collection container, a luminescence reagent injection unit for injecting a luminescence reagent into the washed second collection container, and a luminescence unit. A reaction measurement unit, and the control unit detects that the sample solution has been filtered by the filtration unit, and moves the filtration membrane to the concentrate generation unit by the moving device to concentrate the microorganism. Producing a liquid, detecting that the concentrated liquid has been injected into the first recovery container, and generating ultrasonic waves. , The end of the ultrasonic crushing is detected, the first nozzle is operated to inject the reaction solution in which the antigen is diffused into the second collection container, and then the shaker is operated. After performing the first reaction, detecting the end of the first reaction and operating the labeled antibody injection part, operating the shaker again to perform the second reaction and detecting the end of the second reaction The second reaction vessel is moved to the luminescence reaction device by a moving device, and after the second collection container is washed by the washing unit, the luminescence reagent injection unit is operated to put the luminescence reagent into the second collection vessel. An immunoassay device for automating immunoassay of the microorganism by injecting and further operating a luminescence reaction measurement unit to measure a luminescence reaction, wherein the microorganism collection member according to claim 1 is provided. And in the first collection container, Provided is an immunoassay device characterized in that a bead body of the microorganism collection member is brought into contact with the filtration membrane in the reaction solution to mix the microorganism and the reaction solution.

  The invention according to claim 4 is a collection unit for collecting the immunoassay target microorganism in a reaction solution from a sample solution containing the immunoassay target microorganism, and an ultrasonic fragmentation of the collected microorganism to obtain an antigen in the microorganism. A pretreatment device having an ultrasonic crushing part that diffuses the reaction solution into the reaction solution, an immunoreaction device that causes an antigen-antibody reaction between the antigen and the antibody in the reaction solution, and an antigen-antibody complex produced by the immune reaction A luminescence reaction device that reacts with a luminescent reagent, a device moving device used for immunoassay, and a control unit that controls each device to automate the immunoassay of the microorganism, and the recovery unit includes the A filtration unit that filters the sample liquid and captures the microorganisms in a filtration membrane; and a concentrated solution generation unit that diffuses the microorganisms captured by the filtration membrane into the reaction solution to generate a concentrate, and The ultrasonic crushing part is the concentrated liquid A first recovery container to be injected; and an ultrasonic generator. The immune reaction device includes a second recovery container in which an antibody is fixed to an inner wall thereof to act as a solid phase antibody, and an ultrasonic crushing unit. A nozzle part for sucking a predetermined amount of the reaction solution in which the antigen produced in step 1 is diffused and injecting it into the second recovery container; a labeled antibody injection part for injecting the labeled antibody into the second recovery container; And the luminescence reaction apparatus includes a washing unit for the second collection container, a luminescence reagent injection unit for injecting a luminescence reagent into the washed second collection container, and a luminescence unit. A reaction measurement unit, and the control unit detects that the sample solution has been filtered by the filtration unit, and moves the filtration membrane to the concentrate generation unit by the moving device to concentrate the microorganism. Producing a liquid, detecting that the concentrated liquid has been injected into the first recovery container, and generating ultrasonic waves. , The end of the ultrasonic crushing is detected, the first nozzle is operated to inject the reaction solution in which the antigen is diffused into the second collection container, and then the shaker is operated. After performing the first reaction, detecting the end of the first reaction and operating the labeled antibody injection part, operating the shaker again to perform the second reaction and detecting the end of the second reaction The second reaction vessel is moved to the luminescence reaction device by a moving device, and after the second collection container is washed by the washing unit, the luminescence reagent injection unit is operated to put the luminescence reagent into the second collection vessel. 3. An immunoassay apparatus for automating the immunity measurement of the microorganism by injecting and further operating the luminescence reaction measurement section to measure the luminescence reaction, wherein the microorganism collection member according to claim 2 is provided. And in the first collection container, Provided is an immunoassay device characterized in that the microorganism and the reaction solution are mixed by bringing the dispersion plate of the member for collecting microorganisms into contact with the filtration membrane in the reaction solution.

  In the microorganism collecting member according to the first aspect, since the plurality of bead bodies for dispersing the microorganisms and the holder portion are connected by the string portion, the bead body is removed together with the holder portion. Accordingly, the complicated operation of manually removing each of the beads as in the prior art becomes unnecessary.

  In the microorganism collecting member according to claim 2, since the dispersion plate that disperses the microorganisms in contact with the filter in which the microorganisms are captured is used, the microorganisms can be easily removed, such as a bead removing operation. Complex work is eliminated.

  In the immunoassay device according to claim 3, since the microorganism collecting member according to claim 1 is used, the microorganism collecting step, which has been difficult to be automated in the past, is automated.

  In the immunoassay apparatus according to the fourth aspect, since the microorganism collecting member according to the second aspect is used, the microorganism collecting process that has been difficult to automate conventionally is automated.

  As described above, according to the present invention, the following effects can be obtained.

  Conventionally, it has been necessary to take out beads one by one when collecting microorganisms. However, with the pickup beads according to the present invention, since beads can be put in and out together, it is easy to collect microorganisms. It can be carried out.

  Furthermore, the microorganism collection member according to the present invention can easily collect microorganisms without using beads themselves.

  It is possible to automate the microorganism recovery process, which has been difficult to automate in the past, with a simple configuration and at low cost.

  It eliminates the deterioration of measurement accuracy and manual labor caused by accidental errors and systematic errors in manual immunoassay, and enables quantitative measurement of the amount of antigen in solution and high-precision quantitative measurement of the amount of antibody in solution. .

Example 1
FIG. 1 shows the main configuration of the immunoassay apparatus according to this example. The moving device and the control device are not shown.

  This immunoassay device includes a pretreatment device 106 including a filtration unit 101, a concentrate generation unit 102, and an ultrasonic crushing unit 105, and an immune reaction device 111, a luminescence reaction device 116, and a movement not shown. Device and controller.

  The ultrasonic crushing unit 105 includes a first collection container 103 and an ultrasonic generation unit 104, and the immune reaction device 111 includes a nozzle unit 107, a labeled antibody injection unit 108, a shaking unit 109, and a second collection container 110. An antibody is fixed to the inner wall of the second collection container 110 and acts as a solid phase antibody.

  The luminescence reaction device 115 includes a cleaning unit 112, a luminescence reaction measurement unit 113, and a luminescence reagent injection unit 114.

  Hereinafter, a specific operation of the control unit in this immunoassay device will be described.

1. Operation in Pretreatment Device 106 It is detected that the sample solution is set in the filtration unit 101, the filtration unit 101 is operated, and microorganisms in the sample solution are captured on the filtration membrane (filter). Then, the completion | finish of filtration is detected and the concentrate production | generation part 102 is act | operated. The operated concentrated liquid generating unit 102 generates a concentrated liquid of microorganisms and injects the generated concentrated liquid into the first recovery container 103.

  After detecting that the concentrate has been injected, the ultrasonic generator 104 is operated to perform ultrasonic crushing.

  Furthermore, the end of ultrasonic crushing is detected, the moving device is controlled to move the nozzle unit 107 into the first collection container 102, and the concentrated liquid after ultrasonic crushing (hereinafter referred to as reaction liquid) is sucked. After that, it is moved into the second recovery container 110 and the reaction solution is injected into this recovery container.

2. Operation in the immune reaction device 111 It is detected that the reaction solution has been injected into the second collection container 110, the shaking unit 109 is activated, and the antigen in the reaction solution and the antibody fixed on the inner wall of the second collection container Are reacted (first reaction).

  Next, after detecting the end of the first reaction, the labeled antibody injection unit 108 is operated to inject the labeled antibody into the second collection container 110, and the shaking unit 109 is operated again to advance the antigen-antibody reaction. (Second reaction).

3. Operation in Luminescent Reaction Device 116 It is detected that the second reaction has been completed, and the second recovery container 110 is moved to the washing unit 112 by the moving device, and the reaction solution is removed and washed. Then, the second collection container is moved to the luminescence reaction measurement unit 113.

  Next, the luminescence reaction reagent injection unit 115 is operated to inject the luminescence reagent into the second recovery container 110 and the luminescence reaction measurement unit 113 measures the luminescence reaction.

  By performing the control in this way by the control device, the immunoassay can be performed quickly and with high accuracy.

Example 2
In this embodiment, details of the filtration unit in the pretreatment apparatus will be described.

  FIG. 2 shows the filtration unit.

  The filter 201 and the suction flask 205 are connected with a pipe. The flask can be evacuated by a vacuum pump (not shown). A filter membrane 202 (filter) is set in the filter, and a sample solution to be measured is poured.

  Filtration starts when the cock 206 is opened (the mesh size of the filter is selected to be smaller than the measurement substance). In the immunoassay device, the operation of these decompression pumps and the opening / closing of the cock 206 are controlled by a control device.

When a human performs a filtration operation, after confirming that the filtration has been completed, the filter is transferred to a beaker, and glass beads 203 and a small amount of liquid are added and shaken with a shaker 204. The measurement substance adhering to the surface of the filter is rubbed with a glass beaker and released from the filter surface and dispersed in the liquid. By collecting this solution, a concentrated solution of the measurement substance can be obtained. (Reference: Nihon Millipore Limited, Simplified Microorganism Testing with Membrane Filters, Technical Literature (1992), Akiyoshi Yamaguchi, General Microbiology, Gihodo p15 (1968)
However, when performing automatic measurement with an immunoassay device, the filter (filtration membrane) must be held and moved without using human power.

  In this embodiment, the filter is moved to a predetermined place by holding the filter by the filter holding device (filter adsorber) and moving the filter holding device by the moving device. The structure of this filter holding device is shown in FIG. 3, FIG. 4, and FIG. 3 is a cross-sectional view taken along ABC in FIG. 4, and FIG. 5 is a view of FIG. 3 viewed from the right lateral direction.

  In this filter holding device, the filter is sucked and moved with a negative pressure.

  Reference numeral 311 denotes a connector hole for connecting a pipe for sucking air with a connector, and the air enters from the air introduction hole 13 which is open on the circumference around the point B and enters the ventilation opening 12. Further, it is connected to a decompression pump (vacuum pump) at the end of a pipe connected by a connector (not shown).

  Further, a pickup receiver 16 is provided in the filter holding device so that the moving device can easily hold the filter holding device. These are connected by a plate 15.

  When capturing the filter, the filter is adsorbed on the D-D 'plane. The D-D ′ plane has the shape shown in FIGS. 6, 7, and 8. In the shape of FIG. 6, 20 introduction holes 613 having a diameter of 1 mm are formed. In the shape of FIG. 7, 20 semicircular introduction holes 713 having a radius of 1.5 mm are formed. In the shape of FIG. 8, a groove 813 having a width of 1 mm is formed on the circumference to serve as an air introduction hole.

  The filter holding device of FIG. 3 adopts the shape of FIG. 6, and FIG. 9 is an enlarged view of the air introduction hole portion.

  FIG. 10 is a cross-sectional view taken along the line E-E ′ of FIG.

  First, the air inlet holes having the three shapes shown in FIGS. 6, 7, and 8 were compared using a filter having a diameter of 47 mm and a thickness of 0.4 μm.

  As a result of the experiment, the filter capture / release was most stable in the shape of FIG.

  FIG. 10 shows a state where the filter is adsorbed. On the other hand, as shown in FIG. 11, the measurement substance 1101 adheres to the filter surface after the filtration operation. The filter holding device is provided with a space 14 for keeping a certain distance from the filter so as not to directly touch these substances.

  Also, a pickup receiver 16 for holding the pickup is prepared so that the pickup of the mobile device can be easily held. With such a filter holding device, it is possible to hold the filter as it is without touching the substance captured by the filter.

  In particular, in the immunoassay device, the filter holding device is controlled by the control device to capture and release the filter, and the moving device is controlled to move the filter holding device by the moving device, thereby moving the filter and Capture and release can be performed automatically without using human power.

Example 3
Device configurations of the immune reaction device and the luminescence reaction device are shown in FIGS. 12-19, 1 is a test tube stand (recovery container storage unit), 2 is a test tube (recovery container), 3 is a turntable (shaking device), 4 is a test tube moving device (moving) (Device; test tube moving robot) 5 is a temperature controller, 6 is a dispensing device (luminescent reagent injection unit), 7 is a valve, 8 is a nozzle and nozzle base, 9 is a cleaning unit, 10 is a dark box, and 11 and 12 are shutters. , 13 is a sensor, and 14 is a control device.

  The dark box 10, the shutters 11 and 12, and the sensor 13 constitute a light emission reaction device.

  In FIG. 12, a predetermined amount of a solution containing the antigen to be measured is dispensed into a test tube 2 (not shown) that also serves as a reaction vessel to which a solid phase antibody is bound, and placed in a test tube stand that is not shown. The test tube holder is placed in the test tube holder 1. Thereafter, the following immunoassay operation is automatically performed by the immunoassay device.

  The test tubes 2 placed in the test tube stand 1 are carried and installed one by one on the turntable 3 by the moving device 4 shown in FIG. This operation is performed by the control device.

  FIG. 13 shows a top view of the turntable 3. As shown in this figure, a plurality of test tubes 2 can be set up in this test tube stand.

  FIG. 14 is an explanatory diagram of a turntable on which the test tube 2 is installed. Each test tube 2 installed on the turntable 3 is incubated for a certain time (first reaction). During this incubation, each test tube 2 is shaken by repeating normal rotation and reverse rotation of the turntable 3 by a driving device (not shown).

  At this time, as shown in FIG. 14, the temperature controller 5 built in the turntable 2 is operated to keep the temperature of each test tube 2 constant. If the temperature at the time of reaction is different for each measurement, the measurement system becomes low. Shaking and temperature control of these test tubes are performed by a turntable controller.

  After completion of the incubation, the dispensing device 6 shown in FIG. 17 and the valve 7 shown in FIG. 5 are operated to dispense the labeled antibody solution from the nozzle 8.

  Similarly to the first reaction, each test tube 2 placed on the turntable 3 is incubated again for a certain time (second reaction).

  After completion of the second reaction, each test tube 2 is washed one by one by the washing section 9 shown in FIG.

  A luminescent reagent solution is dispensed into each test tube 2 after washing in the same manner as the labeled antibody. The reagent injection, reagent injection, and incubation are performed in the reagent injection control unit. Thereafter, the moving device 4 is operated by the control device, and each test tube 2 is moved one by one to the dark box 10 where outside light is blocked. This dark box is shown in FIG.

  As shown in FIG. 18, at the time of measurement, the control unit closes the shutter 11 at the top of the dark box 10 and opens the shutter 12 of the sensor 13 (photomultiplier tube) attached to the side surface of the dark box 10. Measure the amount.

  After measuring the amount of chemiluminescence, the moving device is operated by the control device, and each test tube 2 is returned to the test tube stand placed in the test tube stand 1.

  In this way, a series of these automatic operations are controlled by the control device 14 of FIG. 19, and automatic measurement is performed.

  Next, a configuration example (or modification) of each unit in the immunoassay device will be described.

(A) Modification of Temperature Controller Although the temperature controller 5 shown in FIG. 14 is an example using a heater, cooling may be required depending on the reaction. In this modification, a configuration of a temperature controller using a Peltier module is shown.

  FIG. 20 shows a schematic configuration diagram of the temperature controller. This temperature controller is composed of a Peltier module 15, a cooling fin 16, a motor 17, a fan 18, a temperature sensor 19, and a temperature controller 20, and is installed at the lower part of the test tube installation part 21 of the turntable 3.

  The Peltier module is an element that generates a temperature difference due to a potential difference, and the potential difference is controlled by the temperature controller 20 as follows so that the test tube installation unit 21 has an appropriate temperature.

  The temperature controller 20 measures the temperature of the test tube installation part 21 with the temperature sensor 19, and if it is determined that the temperature is lower than the target value, the potential difference is set so that the test tube installation part 21 side of the Peltier module becomes high temperature. Control.

  When the temperature of the test tube installation unit 21 is higher than the target value, the potential difference is controlled so that the temperature of the test tube installation unit 21 side of the Peltier module becomes low. However, when the temperature on the cooling fin side of the Peltier module is higher than the target value, the temperature of the test tube installation portion 21 cannot be cooled to the target value even if the potential difference is controlled.

  In this case, the motor 17 is driven to rotate the cooling fin 16 and the fan 18 is driven to cool the cooling fin 16 so that its temperature becomes lower than the target value.

(B) Configuration example of test tube moving device (moving device) and configuration example of test tube (collection container) In this configuration example, the pickup unit is configured by two members (pickup members a and b), It was detected whether or not the test tube was picked up by detecting a certain state (for example, capacitance or resistance value). FIG. 21 shows an example of the configuration.

  In FIG. 21, 2101 is a pickup member a, 2102 is a pickup member b, 2103 is a lead wire attached to the pickup member a, 2104 is a lead wire attached to the pickup member b, 2105 to 2106 are test tubes, 2108 and 2109. Indicates an insulator.

  In this example, the pickup members 2101 and 2102 are insulated when the test tube (collection container) is not picked up.

  Here, a test tube is prepared in which the pickup members a and b are conducted in a state where the test tube is picked up. By using this test tube, the pickup members a and b are used when the test tube is not picked up. Insulated and the pick-up members a and b conduct when the test tube is picked up.

  Therefore, by checking the resistance value between the lead wires 2103 and 2104, it can be easily detected whether or not the test tube has been picked up. In this configuration example, a special test tube is prepared. Even when a normal test tube is used, the lead wire is not picked up when the test tube is picked up and not picked up except for a special case. Since the electrical resistance and capacitance of the pickups are different, the success or failure of the pickup can be detected by detecting the change.

  Moreover, the shape shown in FIG. 23 is mentioned as a shape of a test tube.

  In this configuration example, as shown in FIG. 23 (c), a groove was formed at the top of the test tube so as to pass through the center of the test tube. By forming the groove portion in this way, elasticity is generated in the test tube, and when the pickup member receives a force F that sandwiches the test tube, the circumferential surface bends inward and returns a reaction force to the tip of the pickup member.

  Thus, even if the force F when picking up the test tube becomes stronger than necessary due to the elasticity (play) caused by the outer shape change, the excessive force is absorbed by the outer shape change and the pickup member is hardly deformed. . When a pulse motor is used as the drive source, the force F can be increased by increasing the pulse amount.

  Without this groove, only the elasticity of the material itself can be obtained, and there is almost no play. Therefore, when the force F increases, all of the excessive force is applied to the pickup member, leading to deformation of the pickup member.

  In addition, although the test tube is hollow, when a solid object such as a cylindrical body is to be moved as shown in FIG. 23 (b), a hole is preferably formed vertically on the top. To form grooves. This groove is preferably shorter than the hole.

(C) Configuration example of a shaker When a test tube is shaken with a shaker, it is necessary to detect the position of the test tube after shaking in order to move the test tube. As the detection method, there is a method in which a light shielding plate is attached to the rotating shaft of the motor, and the position of the test tube is detected by detecting the light shielding state by a photosensor (photosensor).

  FIG. 24 shows an example of the position detection mechanism.

  In FIG. 24, 131 is a photosensor, 132 is a disk with a scratch (light shielding plate), 133 is a motor, and 134 is a turntable. As shown in FIG. 14, the disk 132 with scratches transmits only a specific part of light. Further, since the scratched disc 132 is fixed to the rotating shaft of the motor, it rotates together with the motor when shaken.

  Therefore, if it is prepared so that the light beam is transmitted in the initial state before shaking, the motor is stopped in a state where the light beam is transmitted by the photo sensor, so that the rotating base is always stopped at the same position as the initial state. The test tube is also automatically positioned.

  In FIG. 25, the light beam is transmitted in the initial state. However, any configuration that can detect the rotation angle of the motor based on the state of the transmitted light may be used. For example, the light-shielding plate is in a bar shape and the test tube is positioned. The light source from the light source may be blocked in the state, and the light beam may be transmitted through the other state and detected by the photosensor.

(D) Configuration Example of Cleaning Unit The cleaning unit is configured as shown in FIG. In FIG. 26, 151 is a three-way valve, 152 is a needle, 153 is a collection container (in this example, a test tube), 154 is a cleaning liquid supply tube, and 155 is a suction tube.

  In order to clean the test tube, the cleaning liquid is injected into the test tube 153 from the cleaning liquid supply pipe 154 through the needle 152. After the cleaning is completed, the cleaning liquid in the test tube 152 is removed from the suction pipe 155 through the needle 152 as waste liquid.

  However, when the cleaning liquid is separated and sucked using a single needle, the three-way valve to the tip of the needle are contaminated with unreacted substances, and it becomes difficult to perform normal B / F separation. For this reason, the measurement accuracy is lowered.

  In particular, when cleaning a plurality of test tubes, unreacted substances adhering to the needle 152 are dissolved in the cleaning liquid of other test tubes, and the measurement accuracy is lowered.

  In this configuration example, as shown in FIG. 27, a cleaning liquid injection needle 164 and a cleaning liquid suction needle 165 are prepared to perform cleaning liquid injection and cleaning liquid suction, respectively. FIG. 28 shows an enlarged view at the time of cleaning liquid injection and suction.

  Thus, by performing injection and suction of the cleaning liquid using the dedicated needles, contamination of the cleaning liquid injection needle with unreacted substances can be prevented, and the measurement accuracy is also improved.

Example 4
In this example, a microorganism collecting member that facilitates the microorganism collecting process in the immunoassay and is suitable for automation was manufactured. This member collects microorganisms from a filter that has captured microorganisms. Put a filter and a recovery solution (usually a buffer solution) in a recovery container (for example, a beaker), and shake by placing this microorganism recovery member. To collect microorganisms.

  FIG. 29 is a front view of the microorganism collection member. FIG. 29 shows a member for collecting microorganisms. The member for collecting microorganisms includes a holder 31, a bead 32, and a thread 33. A plurality of yarns 33 are bound to the holder 31 and a bead 32 is fixed to the tip of each yarn.

  Microorganisms were collected using this microorganism collecting member. First, the sample solution is filtered under reduced pressure to capture microorganisms in the filter 34. Next, the filter 34 is put into the beaker 35 as shown in FIG.

  Further, the buffer solution and the microorganism collection member are put in, and shaken by the shaker 36, and the microorganisms attached to the filter 35 are washed away and dispersed and collected in the buffer solution. The microorganism collection member is removed after the microorganisms are collected in the buffer solution.

  Conventionally, beads are separated one by one and cannot be removed at once. When a buffer solution that collects microorganisms is sucked with a nozzle or the like with these beads in place, the nozzle 37 may come into contact with the beads as shown in FIG. The solution that could not be sucked in may remain inside.

  For this reason, it is necessary to remove the beads manually. Conventionally, this process is performed by manually removing the beads one by one, and a very complicated operation is required.

  When the microorganism collecting member as in this embodiment is used, all the beads can be removed together with the holder 31. Therefore, this process can be easily automated.

  FIG. 31 shows a modification of the microorganism collection member. In this microorganism collection member, a shaft 202 is provided on a pickup receiver (holder) 201, and a perforated disk 204 (dispersion plate) is joined to the shaft 202 via a stopper 203.

  Similarly to the microorganism collection member of FIG. 29, the microorganisms can be collected by shaking the microorganism collection member of FIG.

  In this example, the holder 31 and the pickup receiver 201 are formed in a cylindrical shape so that the pickup corresponding to the finger of the robot can be easily grasped. When performing an automatic measurement with the immunoassay device, the moving device of the immunoassay device grasps the holder 31 or the pickup receiver 201 of the microbe collection member and puts the microbe collection member into and out of the beaker.

  In the case of automating the measurement, the sample solution and the buffer solution are sucked and injected by a nozzle or the like. Therefore, it is preferable that the holder portion of the nozzle has a cylindrical shape like the holder portion of the microorganism collection member. A sectional view of such a nozzle and a sectional view of a pickup of the moving device are shown in FIGS. 32 (a) and 32 (b), respectively.

  In FIG. 32A, the nozzle 37 and the holder 39 are fixed by a fixing base 38, and the holder 39 has the same shape as the holder 31 of the microorganism collection member. In FIG. 32B, reference numeral 40 denotes a pickup unit of the moving device. In this way, by unifying the shapes of the holder 31 of the microorganism collection member and the holder 39 of the nozzle, the solution can be collected and injected by a common moving device.

Example 5
In this example, an immunoassay container that facilitates aspirating and collecting a solution in an immunoassay and suitable for automation was manufactured. This immunoassay container can be applied to the first recovery container, the second recovery container, and the like in the first embodiment.

  In order to perform this immunoassay, the handling of the solution is important. Particularly when the solution is sucked and collected, it is required to minimize the amount of the remaining liquid that cannot be collected in the container.

(Α)
First, an example is shown in which when a solution is sucked and collected with a nozzle, the container is tilted to facilitate suction and collection. FIG. 33 is an explanatory diagram thereof. As shown in this figure, by tilting the container, the solution concentrates on the end of the bottom surface of the container and the liquid level rises. When the container is not tilted, the amount of residual liquid increases in proportion to the bottom area of the container, but the amount of residual liquid can be reduced by tilting the container in this way.

  For example, in the case of sucking and collecting the solution after the collection of the microorganisms, the roller 36 is provided in the shaker 36 as shown in FIG. 34 and the installation part of the shaker is inclined to complete the collection of the microorganisms. The container 35 is automatically tilted by moving the shaker later. Even when immunoassay is automated, such a shaker can be easily moved, and the measurement accuracy can be improved with a simple configuration.

(Β)
In the example (α), it is necessary to install the rollers 41 on the shaker and move the shaker. In this example, an immunoassay container that does not require the movement of a special member or a shaker as described above was manufactured. An example is shown in FIG. FIG. 35 (a) is a top view of the immunoassay container according to this example, and FIG. 35 (b) is a cross-sectional view taken along line AA ′.

  As shown in FIG. 35 (b), in this immunoassay container, the cylindrical surface 42 is provided with an inclination angle and a bottom surface 43 is provided, and the same effect as when the container is inclined as in (α) is obtained. can get.

  This immunoassay container is obtained, for example, by preparing a cylinder that forms the wall of the container and a bottom plate that forms the bottom of the container, and inclining and joining the bottom of the container.

  Furthermore, if the outer diameter D1, inner diameter D2 and height H of the immunoassay container are determined, the amount of residual liquid can be minimized by positioning the nozzle 37 at the bottom of the bottom surface.

(Γ)
In this example, a recess 44 having a diameter enough to insert a nozzle is formed on the bottom surface 45 of the immunoassay container, and the nozzle is inserted into the recess 44 to suck the solution. An example is shown in FIG. FIG. 36 (a) is a top view of the immunoassay container, and FIG. 25 (b) is a cross-sectional view taken along line AA ′.

  As shown in FIG. 36 (b), in this immunoassay container, when the amount of the solution in the container decreases, the solution concentrates in the recess 44 and the remaining liquid remains in the recess 44. As described above, the amount of residual liquid is proportional to the bottom area of the solution, but in this container, the bottom area of the recess is very small. Therefore, the amount of the residual liquid becomes very small compared with the residual liquid of a normal container.

  In the case of automating immunoassay, the outer diameter D1, inner diameter D2, height H of the immunoassay container and the distance R between the center of the recess 44 and the center of the container are measured, and the nozzle is the center of the recess. The amount of residual liquid can be minimized by positioning so as to come to the point.

  In particular, as shown in FIG. 37, the bottom surface of the immunoassay container is inclined so that the concave portion 44 becomes the lowermost portion, so that no solution remains in the portion other than the concave portion 44 of the container. it can. In this example, the concave portion is provided at the end of the container. However, the position of the concave portion is not limited as long as the concave portion is inclined so as to be at the lowermost portion. For example, the center of the container may coincide with the center of the concave portion. .

Example 6
The conventional coliform group test has a long measurement time and is difficult to apply to chlorine injection amount control in a sewage treatment plant.

  Therefore, we performed a coliform test by immunoassay and verified the correlation with the conventional plate culture method (deso method) using desoxycholate medium. The correlation coefficient (r) was 0.88 ( n = 182 samples).

  As described above, in the deso method, the measurement takes about 20 to 30 hours, but in this immunoassay method, the measurement can be performed in about 2 hours.

  Since the immunoassay apparatus having the configuration of Example 1 is compatible with this immunoassay method capable of measuring in a short time, the immunoassay can be performed in a short time. Example 5 shows a specific example of the immunoassay. In addition, the apparatus, member, apparatus, etc. which are used for an immunoassay apparatus use what was shown in Examples 2-4.

  A specific processing procedure in the preprocessing apparatus will be described with reference to the flowchart of FIG. In the following flowcharts, all control processes such as operation and stop of the apparatus are performed by the control apparatus, and all movement processes are performed by the movement apparatus controlled by the control apparatus. The moving device shown in Example 3 (B) was used.

  First, a filtration membrane and a sample solution are set. This is done manually.

  In order to increase the filtration rate, a vacuum pump for sample suction in the concentrator is activated (S1).

  The sensor checks the remaining amount of the sample (S2).

  It is detected that there is no remaining sample (S3), and the operation of the vacuum pump is stopped (S4).

  The filtration membrane is captured by the filtration membrane holding device, the filtration membrane holding device is moved into the container in the concentrated liquid generation unit by the moving device, the filtration membrane is released and moved into the container (S5).

  Buffer 2 (ml) is poured into this container (S6), and the microorganism collection member shown in Example 3 is set by the moving device (S7).

  The operation of the shaker shown in Example 3 (C) is started (S8). The shaking time is detected (S9), and the shaker is stopped after a predetermined time (about 5 minutes) (S10).

  The reaction solution in which the coliforms are concentrated is dispensed in an ultrasonic crushing container (first recovery container) by 0.5 (ml) (S11).

  The ultrasonic wave generator is actuated, and the coliform group is crushed by repeating 10 cycles of 30 seconds of ultrasonic irradiation and 60 seconds of rest (S12). The end of crushing is detected (S13).

  In addition, the reaction temperature during ultrasonic crushing is adjusted with the temperature controller (Peltier module) of Example 3 (A). This control was also performed by the control device.

  Next, a specific processing procedure in the immune reaction device will be described with reference to the flowchart of FIG.

  First, the nozzle part is operated to inject 0.5 (ml) of the ultrasonically crushed reaction liquid into the second collection container (S14).

The first reaction is performed by operating the shaker. (S15)
The end of the first reaction is detected (S16), and the operation of the shaker is stopped (S17).

  Reaction buffer 0.5 (ml) is injected into the second collection container (S18). Thereafter, the labeled antibody injection part is operated to dispense 0.1 (ml) of enzyme labeled antibody into the second collection container (S19).

  A shaker is operated and 2nd reaction is performed (S20). The end of the second reaction is detected (S21), and the shaker is stopped (S22).

  Similar to the pretreatment device, the reaction temperature during ultrasonic crushing is adjusted by the temperature controller (Peltier module) of Example 3 (A).

  A specific processing procedure in the luminescence reaction apparatus will be described with reference to the flowchart of FIG. In addition, the luminescent reaction measurement part was performed using the dark box which has the sensor shown by FIG.

  The second collection container is moved to the cleaning unit by the moving device (S23).

  The washing apparatus shown in Example 3 (D) is operated to remove the reaction liquid in the second reaction vessel by suction, and further, the washing process of injecting 5 ml of distilled water to remove it is repeated 5 times ( S24).

  The luminescent reagent injection part is operated to inject luminescent reagent 1 (ml) into the second collection container (S25).

  The shutter 11 of the light emission measuring unit in FIG. 18 is opened (S26), and the second collection container is moved to a predetermined position in the dark box (S27).

Closing the shutter 11 The shutter 12 installed in front of the photomultiplier tube (sensor) is opened (S29).

  The light emission amount is measured and the light emission amount per second is output. This is performed for 30 seconds (S30).

  The end of the light emission measurement is detected and the measurement of the light emission is stopped (S31). The shutter 12 is closed (S32). The shutter 11 is opened (S33).

  The second collection container is removed from the dark box (S34).

  As described above, the immunity is automatically measured without using human power by controlling each unit by the control device. In each of the above examples, a common moving device is used for moving all devices, members, etc., and all the objects to be moved are provided with a holder portion, and this holder portion is moved by the pick-up portion of the moving device. I am doing so. Therefore, it is not necessary to prepare a plurality of moving devices for each moving object, the configuration is simplified, and the cost can be kept low.

  As described above, according to the above embodiment, the following effects can be obtained.

  [1] When the liquid in the container is sucked and collected, the amount of the remaining liquid that cannot be collected can be reduced, and the measurement accuracy can be improved.

  [2] Conventionally, when collecting microorganisms, it was necessary to take in and out the beads one by one. However, in the pick-up beads according to the present invention, since the beads can be put in and out together, Can be recovered.

  Furthermore, the microorganism collection member according to the present invention can easily collect microorganisms without using beads themselves.

  [3] The microorganism recovery process, which has been difficult to automate in the past, can be automated with a simple configuration and at low cost.

  [4] Eliminates measurement accuracy deterioration and manual labor caused by accidental errors and systematic errors in manual immunoassay, quantitative measurement of the amount of antigen in solution, and highly accurate determination of the amount of antibody in solution Can measure.

  [5] By adjusting the temperature during the antigen-antibody reaction, variations in measurement due to the reaction temperature can be suppressed. In particular, by using a Peltier module that generates a temperature difference by a potential difference, the reaction temperature can be easily performed at an appropriate temperature with a small apparatus configuration, and the measurement accuracy is increased.

  [6] An electric resistance value (or capacitance value) between the two pickup members is detected, and whether or not the value is a value in a state where the two pickup members sandwich the storage container is detected. This makes it possible to easily and accurately determine whether or not the storage container is held by the moving device, and the cost is low.

  [7] Since the positioning is performed by the light shielding plate and the photosensor (optical sensor), the positioning can be accurately performed at low cost.

  [8] Since the filter holding device that sucks and holds the entire filter is obtained, the movement from the predetermined position to the predetermined position can be easily performed. For example, from the filter location, the filter can be placed in place on the filter, or the filter can be accurately placed on the bottom of the beaker. Accordingly, the positioning accuracy of the filter is increased.

  In particular, when moving the filter, considering the case of moving the filter with tweezers that can move while the filter filtration surface is kept horizontal, the filter surface becomes perpendicular to the ground when the filter is picked up at one place. There is also a possibility that the substance adhering to the filter filtration surface falls. In contrast, if you are moving horizontally, you don't have to worry.

1 is a schematic configuration diagram of an immunoassay apparatus according to one embodiment of the present invention. Explanatory drawing of a filtration part. Explanatory drawing of a filter holding | maintenance apparatus. The top view of a filter holding | maintenance apparatus. The side view of a filter holding | maintenance apparatus. Explanatory drawing of the filter adsorption | suction part of a filter holding | maintenance apparatus. Explanatory drawing of the filter adsorption | suction part of a filter holding | maintenance apparatus. Explanatory drawing of the filter adsorption | suction part of a filter holding | maintenance apparatus. The enlarged view of an adsorption | suction part. Explanatory drawing of a filter adsorption state. Explanatory drawing of a filter. 1 is a schematic configuration diagram of an immunoassay apparatus according to one embodiment of the present invention. The top view of the turntable of the immunoassay apparatus which concerns on one Example of this invention. The right view of the heater of the immunoassay apparatus which concerns on one Example of this invention. The left view of the immunoassay apparatus which concerns on one Example of this invention. 1 is a front view of an immunoassay device according to one embodiment of the present invention. BRIEF DESCRIPTION OF THE DRAWINGS FIG. 1 is a longitudinal sectional view of a test tube cleaner of an immunoassay apparatus according to one embodiment of the present invention. The longitudinal cross-sectional view of the dark box and sensor of the immunoassay apparatus which concerns on this invention. The right view of the immunoassay apparatus which concerns on one Example of this invention. Explanatory drawing of the temperature regulator which concerns on one Example of this invention. Explanatory drawing of the test tube moving apparatus which concerns on one Example of this invention. Explanatory drawing of the test tube moving apparatus which concerns on one Example of this invention. Explanatory drawing of the test tube moving apparatus which concerns on one Example of this invention. Explanatory drawing of the shaker which concerns on one Example of this invention Explanatory drawing of the light-shielding plate which concerns on one Example of this invention. Explanatory drawing of a test tube cleaning apparatus. Explanatory drawing of the test tube cleaning apparatus which concerns on one Example of this invention. The enlarged view of the test-tube washing | cleaning apparatus which concerns on one Example of this invention. Explanatory drawing of the member for microorganism collection which concerns on one Example of this invention. Explanatory drawing of the microorganisms collection process which concerns on one Example of this invention. Explanatory drawing of the modification of a microorganisms collection member. Explanatory drawing of the pickup which concerns on one Example of this invention. Explanatory drawing of the solution collection | recovery method concerning one Example of this invention. Explanatory drawing of the solution collection | recovery method concerning one Example of this invention. Explanatory drawing of the container for immunoassay which concerns on one Example of this invention. Explanatory drawing of the container for immunoassay which concerns on one Example of this invention. Explanatory drawing of the container for immunoassay which concerns on one Example of this invention. The flowchart which shows a pre-processing process. The flowchart which shows an immunoassay process. The flowchart which shows a light emission measurement process. Explanatory drawing of an antigen antibody reaction. Explanatory drawing of the microorganism concentration process which concerns on a prior art example. Explanatory drawing of the container for immunoassay which concerns on a prior art example. Explanatory drawing of the container for immunoassay which concerns on a prior art example.

Explanation of symbols

DESCRIPTION OF SYMBOLS 1 ... Test tube standing place 2 ... Test tube 3 ... Turntable 4 ... Test tube moving robot 5 ... Heater 6 ... Dispensing device 7 ... Valve 8 ... Nozzle and nozzle stand 9 ... Test tube washing machine 10 ... Dark box 11 ... Shutter 12 ... Shutter 13 ... Sensor 14 ... Control device 101 ... Filtration part 102 ... Concentrated liquid production part 103 ... First collection container 104 ... Ultrasonic wave generation part 105 ... Ultrasonic crushing part 106 ... Pretreatment device 107 ... Nozzle part 108 ... Mark Antibody injection unit 109 ... Shaking unit 110 ... Second collection container 111 ... Immune reaction device 112 ... Cleaning unit 113 ... Luminescence reaction measurement unit 114 ... Luminescence reagent injection unit 115 ... Luminescence reaction device

Claims (4)

A member for collecting microorganisms that mixes microorganisms captured by a filtration membrane with a liquid,
A plurality of beads that disperse the microorganisms in contact with the filter in which the microorganisms are captured;
A holder part;
A member for collecting microorganisms, characterized in that each member is provided on each bead body and has a string portion connecting the holder part and each bead body. A member for collecting microorganisms that mixes microorganisms captured by a filtration membrane with a liquid,
A dispersion plate that contacts the filter in which the microorganisms are captured and disperses the microorganisms;
A disc portion having a holder portion attached in a direction perpendicular to the dispersion plate,
The dispersion plate has a plurality of holes, and the microorganism collecting member. A recovery unit for recovering the immunoassay target microorganism in a reaction solution from a sample solution containing the immunoassay target microorganism, and an ultrasonic disruption for diffusing the antigen in the microorganism into the reaction solution by performing ultrasonic disruption of the recovered microorganism. A pretreatment device having a section,
An immune reaction device that causes an antigen-antibody reaction between the antigen and the antibody in the reaction solution;
A luminescence reaction device for reacting an antigen-antibody complex produced by an immune reaction with a luminescence reagent;
A device for moving instruments used for immunoassay;
A controller that controls each device to automate the immunity measurement of the microorganism,
The recovery unit includes a filtration unit that filters the sample solution and captures the microorganisms in a filtration membrane, and a concentrate generation unit that diffuses the microorganisms captured by the filtration membrane into a reaction solution to generate a concentrate. Have
The ultrasonic crushing unit has a first recovery container into which the concentrated liquid is injected, and an ultrasonic generation unit,
The immune reaction apparatus is configured to aspirate a predetermined amount of a second collection container in which an antibody is fixed on its inner wall and acts as a solid phase antibody, and a reaction solution in which an antigen generated in the ultrasonic crushing portion is diffused. A nozzle part for injecting into the second recovery container, a labeled antibody injection part for injecting the labeled antibody into the second recovery container, and a shaking part of the second recovery container,
The luminescence reaction apparatus has a washing unit for the second collection container, a luminescence reagent injection unit for injecting a luminescence reagent into the washed second collection container, and a luminescence reaction measurement unit,
The controller is
Detecting that the sample liquid has been filtered by the filtration unit, moving the filtration membrane to the concentrate generation unit by the moving device to generate the microorganism concentrate,
Detecting that the concentrated liquid has been injected into the first recovery container and operating the ultrasonic generator,
After the end of the ultrasonic crushing is detected, the first nozzle is operated to inject the reaction solution in which the antigen is diffused into the second recovery container, and then the shaker is operated to perform the first reaction. ,
After detecting the end of the first reaction and activating the labeled antibody injection part, the second reaction is performed by activating the shaker again,
The completion of the second reaction is detected, the second reaction vessel is moved to the luminescence reaction device by a moving device, the second collection vessel is washed by the washing unit, and then the luminescent reagent injection unit is operated to emit light. In the immunoassay device, wherein the immunity measurement of the microorganism is automated by injecting a reagent into the second recovery container and further operating the luminescence reaction measurement unit to measure the luminescence reaction,
A microorganism recovery member according to claim 1, wherein the bead body of the microorganism recovery member is brought into contact with the filtration membrane in the reaction liquid in the first recovery container, and the microorganism and the reaction liquid are contacted. And an immunoassay device characterized by mixing them. A recovery unit for recovering the immunoassay target microorganism in a reaction solution from a sample solution containing the immunoassay target microorganism, and an ultrasonic disruption for diffusing the antigen in the microorganism into the reaction solution by performing ultrasonic disruption of the recovered microorganism. A pretreatment device having a section,
An immune reaction device that causes an antigen-antibody reaction between the antigen and the antibody in the reaction solution;
A luminescence reaction device for reacting an antigen-antibody complex produced by an immune reaction with a luminescence reagent;
A device for moving instruments used for immunoassay;
A controller that controls each device to automate the immunity measurement of the microorganism,
The recovery unit includes a filtration unit that filters the sample solution and captures the microorganisms in a filtration membrane, and a concentrate generation unit that diffuses the microorganisms captured by the filtration membrane into a reaction solution to generate a concentrate. Have
The ultrasonic crushing unit has a first recovery container into which the concentrated liquid is injected, and an ultrasonic generation unit,
The immune reaction apparatus is configured to aspirate a predetermined amount of a second collection container in which an antibody is fixed on its inner wall and acts as a solid phase antibody, and a reaction solution in which an antigen generated in the ultrasonic crushing portion is diffused. A nozzle part for injecting into the second recovery container, a labeled antibody injection part for injecting the labeled antibody into the second recovery container, and a shaking part of the second recovery container,
The luminescence reaction apparatus has a washing unit for the second collection container, a luminescence reagent injection unit for injecting a luminescence reagent into the washed second collection container, and a luminescence reaction measurement unit,
The controller is
Detecting that the sample liquid has been filtered by the filtration unit, moving the filtration membrane to the concentrate generation unit by the moving device to generate the microorganism concentrate,
Detecting that the concentrated liquid has been injected into the first recovery container and operating the ultrasonic generator,
After the end of the ultrasonic crushing is detected, the first nozzle is operated to inject the reaction solution in which the antigen is diffused into the second recovery container, and then the shaker is operated to perform the first reaction. ,
After detecting the end of the first reaction and activating the labeled antibody injection part, the second reaction is performed by activating the shaker again,
The completion of the second reaction is detected, the second reaction vessel is moved to the luminescence reaction device by a moving device, the second collection vessel is washed by the washing unit, and then the luminescent reagent injection unit is operated to emit light. In the immunoassay device, wherein the immunity measurement of the microorganism is automated by injecting a reagent into the second recovery container and further operating the luminescence reaction measurement unit to measure the luminescence reaction,
3. The microorganism-recovering member according to claim 2, wherein a dispersion plate of the microorganism-recovering member is brought into contact with the filtration membrane in the reaction liquid in the first recovery container, thereby the microorganism and the reaction liquid. And an immunoassay device characterized by mixing them.

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