Microorganism detection device and microorganism detection method

Abstract

To enable the detection of a target microorganism with simple operations.SOLUTION: A microorganism detection device comprises: a reaction container that holds therein an adhesion surface to which a test object has been adhered; a blocking processing unit that performs blocking processing on the adhesion surface of the reaction container; a cleaning liquid supply unit that supplies cleaning liquid to the reaction container; a primary antibody solution supply unit that supplies a primary antibody solution containing a primary antibody that captures protein to the reaction container; a secondary antibody solution supply unit that supplies a secondary antibody solution containing a secondary antibody that reacts with the primary antibody and increases the reaction during measurement to the reaction container; a discharge unit for discharging liquid from the reaction container; a detection unit that measures the test object processed in the reaction container and detects a microorganism contained in the test object; and a control unit that executes processing of sequentially supplying and discharging the primary antibody solution and the secondary antibody solution to the reaction container after performing blocking processing on the adhesion surface in the reaction container.SELECTED DRAWING: Figure 1

Description

[Technical Field]

[0001] The present disclosure relates to a microorganism detection device and a microorganism detection method. [Background technology]

[0002] One method for detecting microorganisms is the immunoassay method, which utilizes an antibody-antigen reaction to capture and detect microbial proteins (see Patent Documents 1 and 2). [Prior art documents] [Patent documents]

[0003] [Patent Document 1] Japanese Patent Application Laid-Open No. 2016-99131 [Patent Document 2] Japanese Patent Application Laid-Open No. 2015-222197 Summary of the Invention [Problem to be solved by the invention]

[0004] In Patent Document 1, the microorganisms are crushed to make the proteins easier to detect, and then the proteins are captured by an antigen-antibody reaction, which makes the processing steps complicated.The method described in Patent Document 2 can detect the target microorganisms, etc. with a simple operation, but the targets that can be detected are limited.

[0005] The present disclosure has been made in view of the above-mentioned problems, and aims to provide a microorganism detection device and a microorganism detection method that can detect target microorganisms with a simple operation. [Means for solving the problem]

[0006] In order to achieve the above-mentioned object, the microbial detection device of the present disclosure includes a reaction vessel that holds an adhesion surface to which a test object is attached inside, a blocking processing unit that performs a blocking process on the adhesion surface of the reaction vessel, a cleaning liquid supply unit that supplies a cleaning liquid to the reaction vessel, a primary antibody liquid supply unit that supplies a primary antibody liquid containing a primary antibody that captures protein to the reaction vessel, a secondary antibody liquid supply unit that supplies a secondary antibody liquid containing a secondary antibody that reacts with the primary antibody and increases the reaction during measurement to the reaction vessel, a discharge unit that discharges liquid from the reaction vessel, a detection unit that measures the test object treated in the reaction vessel and detects microorganisms contained in the test object, and a control unit that performs a blocking process on the adhesion surface in the reaction vessel and then executes a process of supplying and discharging the primary antibody liquid and the secondary antibody liquid in that order to the reaction vessel.

[0007] The microbial detection method of the present disclosure for achieving the above-mentioned object includes the steps of: performing a blocking treatment on the surface of a test object held in a reaction vessel and having the test object attached thereto; discharging the cleaning solution and supplying a primary antibody solution containing a primary antibody that captures proteins into the reaction vessel; discharging the primary antibody solution and supplying a cleaning solution; discharging the cleaning solution and supplying a secondary antibody solution containing a secondary antibody that reacts with the primary antibody and increases the reaction during measurement into the reaction vessel; discharging the secondary antibody solution and supplying a cleaning solution; measuring the test object treated in the reaction vessel and detecting microorganisms contained in the test object; and controlling the supply and discharge of liquids from each part, and automatically supplying and discharging the blocking solution, primary antibody solution, and secondary antibody solution to the reaction vessel in that order. [Effects of the Invention]

[0008] According to the present disclosure, target microorganisms can be detected with simple operations. [Brief explanation of the drawings]

[0009] [Figure 1] FIG. 1 is a schematic diagram showing the general configuration of the microorganism detection device of this embodiment. [Figure 2]FIG. 2 is a flowchart showing an example of processing performed by the microorganism detection device. [Figure 3] FIG. 3 is a schematic diagram showing an example of the configuration of a reaction vessel. [Figure 4] FIG. 4 is a schematic diagram showing an example of the configuration of the inspection object acquisition unit. [Figure 5] FIG. 5 is a schematic diagram showing an example of the configuration of the inspection object acquisition unit. [Figure 6] FIG. 6 is a schematic diagram showing an example of the configuration of the inspection object acquisition unit. [Figure 7] FIG. 7 is a schematic diagram showing a schematic configuration of a microorganism detection device according to another embodiment. [Figure 8] FIG. 8 is a flowchart showing an example of processing by the microorganism detection device. [Figure 9] FIG. 9 is a schematic diagram showing an example of a wiping tool. [Figure 10] FIG. 10 is a schematic diagram showing an example of a wiping tool. [Figure 11] FIG. 11 is a schematic diagram showing an example of a wiping tool. [Figure 12] FIG. 12 is a schematic diagram showing an example of a reaction vessel. [Figure 13] FIG. 13 is a schematic diagram showing an example of a reaction vessel. [Figure 14] FIG. 14 is a schematic diagram showing a schematic configuration of a microorganism detection device according to another embodiment. [Figure 15] FIG. 15 is a flowchart showing an example of processing by the microorganism detecting device. DETAILED DESCRIPTION OF THE INVENTION

[0010] Hereinafter, embodiments of the present disclosure will be described in detail with reference to the drawings. However, the present disclosure is not limited to these embodiments. Furthermore, the components in the following embodiments include those that are replaceable and easy for those skilled in the art, or those that are substantially identical. Furthermore, the components in the embodiments described below can be variously omitted, replaced, or modified without departing from the spirit of the present invention. Furthermore, when there are multiple embodiments, the present disclosure also includes those configured by combining the respective embodiments.

[0011] Fig. 1 is a schematic diagram showing the overall configuration of a microorganism detection device according to this embodiment. The microorganism detection device 10 shown in Fig. 1 includes a test object acquisition unit 12, a wiping jig 14, a reaction vessel 16, a blocking processing unit 18, a primary antibody liquid supply unit 20, a secondary antibody liquid supply unit 24, a discharge unit 26, a detection unit 28, and a control unit 30. The microorganism detection device 10 attaches a test object to the wiping jig 14, holds the test object in the reaction vessel 16, and performs blocking processing, primary antibody processing, and secondary antibody processing. After that, the detection unit 28 detects the presence or absence of microorganisms adhering to the test object and the amount of microorganisms. Here, the microorganisms include viruses, bacteria, etc.

[0012] The test object acquisition unit 12 executes a process of attaching the test object to a wiping jig 14. The test object acquisition unit 12 wipes the test object from the object to which it is attached using the wiping jig 14. For example, if the test object is human secretions or tissue, the wiping jig 14 is rubbed against the human to which the test object is attached, causing the test object to attach to the wiping jig 14.

[0013] The wiping jig 14 has an attachment surface 40, a support member 42, and a rod-shaped member 44. The attachment surface 40 is a surface onto which the test object is attached. The attachment surface 40 is a film formed of PVDF (polyvinylidene fluoride) or nitrocellulose. The support member 42 is a plate-shaped member that supports the attachment surface 40. The support member 42 is formed of a material harder than the attachment surface 40. One end of the rod-shaped member 44 is connected to an end of the support member 42. The rod-shaped member 44 is a part that is held by an operator when transporting or operating the wiping jig 14. The operator may be a human or an automatically driven machine. The wiping jig 14 is operated while holding the rod-shaped member 44, and the test object is wiped off with the attachment surface 40 by rubbing the attachment surface 40 against an object using the test object acquisition unit 12, thereby attaching the test object to the attachment surface 40.

[0014] The reaction vessel 16 is a vessel capable of holding the wiping jig 14 therein. The reaction vessel 16 is capable of supplying liquid to the space in which the wiping jig 14 is placed, filling the reaction vessel 16 with liquid, and discharging the filled liquid.

[0015] The blocking processing unit 18 includes a blocking liquid tank 50, a supply channel 52, and an on-off valve 54. The blocking liquid tank 50 stores a blocking liquid used to perform a blocking process on the test object. The blocking liquid is a liquid containing a substance that binds to non-specific sites on the test object, i.e., a substance that inhibits the primary antibody from binding to sites other than the target site. The supply channel 52 is a conduit connecting the blocking liquid tank 50 and the reaction vessel 16. The on-off valve 54 is disposed on the supply channel 52. When opened, the on-off valve 54 supplies the blocking liquid from the blocking liquid tank 50 to the reaction vessel 16, and when closed, the on-off valve 54 stops the supply of the blocking liquid to the reaction vessel 16.

[0016] The primary antibody liquid supply unit 20 includes a primary antibody liquid tank 60, a supply channel 62, and an on-off valve 64. The primary antibody liquid tank 60 stores primary antibody liquid for performing primary antibody treatment on the test object. The primary antibody liquid is a liquid containing a primary antibody that adheres to a specific site on the test object. The primary antibody may have the property of adhering to multiple specific sites. The primary antibody liquid may also contain multiple primary antibodies that each adhere to a different site. This allows multiple proteins to be detected. The supply channel 62 is a conduit connecting the primary antibody liquid tank 60 and the reaction vessel 16. The on-off valve 64 is disposed in the supply channel 62. When opened, the on-off valve 64 supplies the primary antibody liquid from the primary antibody liquid tank 60 to the reaction vessel 16, and when closed, stops the supply of the primary antibody liquid to the reaction vessel 16.

[0017] The secondary antibody liquid supply unit 22 includes a secondary antibody liquid tank 70, a supply channel 72, and an on-off valve 74. The secondary antibody liquid tank 70 stores secondary antibody liquid used for secondary antibody treatment of the test subject. The secondary antibody liquid is a liquid containing a secondary antibody that adheres to a primary antibody. The secondary antibody is a substance that reacts with high sensitivity to detection by the detection unit 28. For example, in the case of analysis using fluorescent light, the secondary antibody is a substance that emits a large amount of light at a predetermined wavelength. The secondary antibody liquid may contain multiple secondary antibodies that each adhere to a different primary antibody. This allows multiple proteins to be detected. The supply channel 72 is a conduit connecting the secondary antibody liquid tank 70 and the reaction vessel 17. The on-off valve 74 is disposed on the supply channel 72. When opened, the secondary antibody liquid is supplied from the secondary antibody liquid tank 70 to the reaction vessel 17, and when closed, the supply of the secondary antibody liquid to the reaction vessel 17 is stopped.

[0018] The cleaning liquid supply unit 24 includes a cleaning liquid tank 80, a supply path 82, and an on-off valve 84. The cleaning liquid tank 80 stores a cleaning liquid for cleaning the reaction vessel 16 in which the test object is placed. The cleaning liquid may be, for example, pure waterFurthermore, different liquids may be supplied as the cleaning liquid depending on the cleaning step. Supply path 82 is a conduit connecting cleaning liquid tank 80 and reaction vessel 16. Opening and closing valve 84 is disposed on supply path 82, and supplies cleaning liquid from cleaning liquid tank 80 to reaction vessel 16, and closing the valve stops the supply of cleaning liquid to reaction vessel 16. Cleaning liquid supply unit 24 cleans reaction vessel 16 by supplying cleaning liquid to reaction vessel 16.

[0019] The discharge unit 26 has a discharge channel 90, a drain tank 92, a recovery channel 94, and on-off valves 96 and 98. The discharge channel 90 is connected to the reaction vessel 16 and receives liquid discharged from the reaction vessel 16. The drain tank 92 is connected to the discharge channel 90. The drain tank 92 stores liquid discharged from the reaction vessel 16. The recovery channel 94 is a conduit connecting the discharge channel 90 with the primary antibody liquid tank 60 and the secondary antibody liquid tank 70. The on-off valve 96 is disposed in the discharge channel 90 and switches between on and off of the path connecting the reaction vessel 16 and the drain tank 92. The on-off valve 98 is disposed in the discharge channel 90 and switches between on and off of the paths connecting the reaction vessel 16 with the primary antibody liquid tank 60 and the secondary antibody liquid tank 70. The on-off valve 98 may be provided for each of the paths connecting the primary antibody liquid tank 60 and the secondary antibody liquid tank 70.

[0020] The detection unit 28 analyzes the test subject processed in the reaction vessel 16 and determines whether the test subject contains a specific protein. The detection unit 28 detects whether the test subject contains a specific protein by detecting the emission of the secondary antibody using, for example, a fluorescent analysis method.

[0021] The control unit 30 controls the operation of each component of the microbial detection device 10. The control unit 30 includes an integrated circuit (processor) such as a CPU or GPU (Graphics Processing Unit), a memory that serves as a working area, and a nonvolatile storage device such as a magnetic storage device or semiconductor storage device, and executes various programs using these hardware resources to perform various processes. The control unit 30 controls the blocking process performed on the test subject by the blocking processing unit 18, the primary antibody process performed on the test subject by the primary antibody solution supply unit 20, the secondary antibody process performed on the test subject by the secondary antibody solution supply unit 22, and the cleaning process performed on the test subject by the cleaning solution supply unit 24. Specifically, the control unit 30 automatically controls the supply and discharge of liquids. Here, the processing of each process may be controlled at a preset time. Furthermore, as will be described later, the control unit 30 may detect the concentration of the liquid and, if a condition is met, proceed to the next process.

[0022] Next, the operation of the microorganism detection device 10 will be described with reference to Fig. 2. Fig. 2 is a flowchart showing an example of processing by the microorganism detection device. Part of the processing shown in Fig. 2 may be performed by an operator. Specifically, the operation of moving the wiping jig may be performed by an operator. All processing may be performed automatically using a machine. The operation of the microorganism detection device 10 will be described below.

[0023] The microorganism detection device 10 prepares a wiping jig (step S12). The microorganism detection device 10 prepares the attachment surface 40 of the wiping jig 14 so that the test subject can be attached to it. For example, the surface of the attachment surface 40 is processed to make it easier for the test subject to attach, and a wetting process and equilibration process are performed.

[0024] Next, the microorganism detection device 10 adheres the test object to the attachment surface (step S14). The microorganism detection device 10 grasps and operates the rod-shaped member 44 of the wiping jig 14, brings the attachment surface 40 into contact with the portion where the test object is placed, and wipes the test object with the attachment surface 40.

[0025] Next, the microorganism detection device 10 performs a blocking process (step S16). The microorganism detection device 10 places the wiping jig 14 with the test object attached in the reaction vessel 16. The microorganism detection device 10 opens the on-off valve 54 of the blocking processing unit 18, supplies a blocking liquid to the reaction vessel 16, and fills the reaction vessel 16 with the blocking liquid. By filling the reaction vessel 16 with the blocking liquid, the microorganism detection device 10 brings the blocking liquid into contact with the test object on the attachment surface 40, and causes the substances in the blocking liquid to react with non-specific parts of the test object, preventing the primary antibody from adhering.

[0026] Next, the microorganism detection device 10 performs a cleaning process (step S18). The microorganism detection device 10 opens the on-off valve 96 of the discharge unit 26 and discharges the blocking liquid in the reaction vessel 16 into the waste liquid tank 92. Thereafter, the microorganism detection device 10 opens the on-off valve 84 of the cleaning liquid supply unit 24 and flows the cleaning liquid into the reaction vessel 16. The microorganism detection device 10 may perform so-called pool rinsing, in which the cleaning liquid is stored in the reaction vessel 16 and then discharged, or may perform running water rinsing, in which the cleaning liquid is supplied to the reaction vessel 16 while being discharged with the on-off valve 96 open. The microorganism detection device 10 discharges the cleaning liquid from the reaction vessel 16 and completes the cleaning process.

[0027] Next, the microorganism detection device 10 performs primary antibody treatment (step S20). The microorganism detection device 10 opens the on-off valve 64 of the primary antibody liquid supply unit 20, supplies the primary antibody liquid to the reaction vessel 16, and fills the reaction vessel 16. By filling the reaction vessel 16 with the primary antibody liquid, the microorganism detection device 10 brings the primary antibody into contact with the test object on the attachment surface 40, and causes the primary antibody to react with a specific portion of the test object, resulting in the primary antibody being attached.

[0028] Next, the microorganism detection device 10 performs a cleaning process (step S22). The microorganism detection device 10 opens the on-off valve 94 of the discharge unit 26 and discharges the primary antibody liquid from the reaction container 16 into the primary antibody liquid tank 60. Thereafter, the microorganism detection device 10 opens the on-off valve 84 of the cleaning liquid supply unit 24 and flows the cleaning liquid into the reaction container 16. The microorganism detection device 10 discharges the cleaning liquid from the reaction container 16 and completes the cleaning process.

[0029] Next, the microorganism detection device 10 performs secondary antibody treatment (step S24). The microorganism detection device 10 opens the on-off valve 74 of the secondary antibody liquid supply unit 22, supplies the secondary antibody liquid to the reaction container 16, and fills the reaction container 16. By filling the reaction container 16 with the secondary antibody liquid, the microorganism detection device 10 brings the secondary antibody into contact with the primary antibody attached to the test object on the attachment surface 40, and causes the secondary antibody to react with the primary antibody attached to a specific portion of the test object, resulting in the secondary antibody being attached.

[0030] Next, the microorganism detection device 10 performs a cleaning process (step S26). The microorganism detection device 10 opens the on-off valve 94 of the discharge unit 26 and discharges the secondary antibody liquid from the reaction container 16 into the secondary antibody liquid tank 70. Thereafter, the microorganism detection device 10 opens the on-off valve 84 of the cleaning liquid supply unit 24 and flows the cleaning liquid into the reaction container 16. The microorganism detection device 10 discharges the cleaning liquid from the reaction container 16 and ends the cleaning process.

[0031] Next, the microorganism detection device 10 processes the specimen object in the detection unit 28 to detect proteins (step S28). The microorganism detection device 10 moves the wiping jig 14 from the reaction container 16 to the detection unit 28 and performs a detection process on the attachment surface. Specifically, it performs a detection process on the secondary antibody attached to the attachment surface 40. The detection unit 28 detects the presence or absence of chemiluminescence from the secondary antibody and the luminescence intensity, thereby detecting the presence or absence and amount of a predetermined protein contained in the detection object.

[0032] The microorganism detection device 10 is provided with a blocking processing unit 18, a primary antibody liquid supply unit 20, and a secondary antibody liquid supply unit 22, and the control unit 30 automatically controls the operation of each unit, making it possible to more easily perform the detection process for the target protein contained in the test object on the wiping jig 14. The microorganism detection device 10 is also provided with a cleaning liquid supply unit 80, and the cleaning process is performed under the control of the control unit 30, which can prevent liquid mixing. This also reduces the burden on the operator. The microorganism detection device 10 can also reduce the amount of liquid used by recovering the primary antibody liquid and the secondary antibody liquid. The microorganism detection device 10 may also recover and reuse the blocking liquid and cleaning liquid. When recovering them, it is preferable to perform a process to remove impurities from the recovered liquid.

[0033] It is preferable that the liquid tanks and reaction vessels in each section are provided with temperature control mechanisms so that the liquid temperatures and processing temperatures can be adjusted.

[0034] FIG. 3 is a schematic diagram showing an example of the configuration of a reaction vessel. The reaction vessel 16a shown in FIG. 3 has a rotation unit 104 and a stirring mechanism 106. The rotation unit 104 is disposed at a portion that supports the rod-shaped member 44 of the wiping jig 14 and rotates the wiping jig 14 relative to the reaction vessel 16a. The rotation unit 104 stirs the liquid in the reaction vessel 16a by rotating the wiping jig 14. The stirring mechanism 106 is a screw or the like disposed inside the reaction vessel 16a. The stirring mechanism 106 stirs the liquid in the reaction vessel 16a. The reaction vessel 16a may also stir the liquid inside by supplying the liquid discharged from the discharge channel 90 to the reaction vessel 16a and circulating the liquid.

[0035] The reaction vessel 16a is provided with a mechanism for stirring the liquid inside, and stirring the liquid can promote contact between the test object attached to the attachment surface 40 and the liquid, thereby promoting reactions in various processes. This can shorten the processing time. Furthermore, it is not necessary to have multiple mechanisms for stirring the liquid, and the reaction vessel 16a may be provided with only one of the rotating unit 104, the stirring mechanism 106, and the mechanism for circulating the liquid.

[0036] FIG. 4 is a schematic diagram showing an example of the configuration of the test object acquisition unit. The housing of the test object acquisition unit 12a shown in FIG. 4 serves as a container for storing the wiping jig 14, with the test object attached to its attachment surface 40, until it is placed in the reaction container 16. The test object acquisition unit 12 supports the rod-shaped member 44 in a state in which the attachment surface 40 of the wiping jig 14 does not come into contact with other parts. This prevents the attachment surface 40 from coming into contact with other parts. A desiccant 110 is disposed inside the housing of the test object acquisition unit 12a. The desiccant 110 is a material that absorbs moisture, such as silica gel.

[0037] By disposing the desiccant 110 in the housing of the inspection object acquisition unit 12a, it is possible to prevent the adhesion surface 40 to which the inspection object is attached from being contaminated by the growth of mold or the like. Furthermore, by supporting the adhesion surface 40 in a state where it does not come into contact with other parts, it is possible to prevent the inspection object acquisition unit 12a, the adhesion surface 40, and the inspection object from being contaminated.

[0038] FIG. 5 is a schematic diagram showing an example of the configuration of the inspection object acquisition unit. The inspection object acquisition unit 12b shown in FIG. 5 has a dry air supply unit 120. The dry air supply unit 120 has a supply pipe 122, an exhaust pipe 124, and a dry air supply source 126. The supply pipe 122 connects the dry air supply source 126 to the housing of the inspection object acquisition unit 12b. The exhaust pipe 124 is connected to the inspection object acquisition unit 12b and is open to the atmosphere. The exhaust pipe 124 may be provided with a check valve to prevent atmospheric air from flowing into the housing of the inspection object acquisition unit 12b. The dry air supply source 126 supplies dry air to the supply pipe 122. The dry air supply unit 126 has a blowing function and a dehumidifying function. The dry air supply unit 120 supplies dry air from the dry air supply source 126 to the housing of the inspection object acquisition unit 12b, thereby keeping the inside of the inspection object acquisition unit 12b dry and preventing the surface of the wiping jig 14 held by the inspection object acquisition unit 12b from becoming contaminated.

[0039] FIG. 6 is a schematic diagram showing an example of the configuration of the inspection object acquisition unit. The inspection object acquisition unit 12c shown in FIG. 6 has a protective liquid supply unit 130. The protective liquid supply unit 130 has a circulation path 132 and a liquid supply source 134. The circulation path 132 connects the housing of the inspection object acquisition unit 12c, which stores the wiping jig 14, to the liquid supply source 134. The liquid supply source 134 supplies a protective liquid that protects the surface of the attachment surface 40, suppresses the growth of mold, etc., and is easy to remove. The protective liquid is ethanol, for example, ethanol with a concentration of 70% or more, or methanol. The protective liquid supply unit 130 supplies the protective liquid from the liquid supply source 134 to the housing of the inspection object acquisition unit 12c, so that the protective liquid is in contact with the surface of the attachment surface 40. When the wiping jig 14 is moved to the reaction container 16, the protective liquid supply unit 130 recovers the protective liquid in the liquid supply source. In this way, instead of drying, a protective liquid can be supplied to prevent contamination of the adhering surface 40 of the wiping tool 14.

[0040] In the above embodiment, the blocking process is performed by supplying a blocking liquid to the attachment surface 40, but the blocking process is not limited to a process using a blocking liquid. Fig. 7 is a schematic diagram showing the general configuration of a microorganism detection device of another embodiment. Of the microorganism detection device 10a shown in Fig. 7, detailed description of the configuration that is similar to the microorganism detection device 10 will be omitted, and only points unique to the microorganism detection device 10a will be described.

[0041] The microorganism detection device 10a includes a test object acquisition unit 12, a wiping jig 14, a reaction container 16, a blocking processing unit 50a, a primary antibody liquid supply unit 20, a secondary antibody liquid supply unit 24, a discharge unit 26, a detection unit 28, and a control unit 30. The blocking processing unit 50a dries the surface of the attachment surface 40 instead of supplying a blocking liquid.

[0042] The blocking treatment unit 50a has a circulation path 200, an on-off valve 202, and a dry air supply source 204. The circulation path 200 is a pipe line connected to the reaction vessel 16 at both ends. The on-off valve 202 is disposed in the circulation path 200 and is a valve that switches the flow of air through the circulation path 200. The dry air supply source 204 has a blowing function and a dehumidifying function, and supplies dry air to the circulation path 200.

[0043] The blocking processing unit 50a supplies dry air from the dry air supply source 204 to the reaction vessel 16 via the circulation path 200, thereby drying the surface of the attachment surface 40 of the wiping jig 14 placed in the reaction vessel 16. This allows the inspection object to be dried. The dry air supply source 204 may be a mechanism that supplies heated air.

[0044] Fig. 8 is a flowchart showing an example of processing performed by the microorganism detecting apparatus. Of the processing shown in Fig. 8, detailed description of the same processing as that shown in Fig. 2 will be omitted.

[0045] The microorganism detecting device 10a prepares a wiping tool (step S12). Next, the microorganism detecting device 10a attaches the inspection target to the attachment surface (step S14).

[0046] Next, the microorganism detection device 10a performs a drying process (step S42). The microorganism detection device 10a places the wiping jig 14 with the test object attached in the reaction container 16. The microorganism detection device 10a supplies dry air from the blocking processing unit 50a to the reaction container 16, and fills the reaction container 16 with a dried firearm. The blocking processing unit 50a circulates the dry air through the circulation path 202, thereby reducing the humidity inside the reaction container 16.

[0047] Next, the microorganism detecting device 10a performs a primary antibody treatment (step S20). Next, the microorganism detecting device 10a performs a cleaning treatment (step S22). Next, the microorganism detecting device 10a performs a secondary antibody treatment (step S24). Next, the microorganism detecting device 10a performs a cleaning treatment (step S26). Next, the microorganism detecting device 10a processes the specimen object in the detection unit 28 to detect proteins (step S28).

[0048] The microorganism detection device 10a can perform the blocking process without using a blocking liquid by drying the attachment surface 40 of the wiping jig 14. This eliminates the need to prepare a blocking liquid, simplifying the process.

[0049] The wiping jig 14 can have a variety of structures. Fig. 9 is a schematic diagram showing an example of a wiping jig. The wiping jig 14a shown in Fig. 9 has attachment surfaces 150a and 150b provided on two opposing surfaces of a support member 152. This allows one wiping jig to be provided with two attachment surfaces 150a and 150b.

[0050] FIG. 10 is a schematic diagram showing an example of a wiping jig. The wiping jig 14b shown in FIG. 10 has a square cross section perpendicular to the extension direction of the rod-shaped member 164 of the support member 162. The wiping jig 14b has adhesive surfaces 160a, 160b provided on four faces of the quadrangle of the support member 162. Although only two faces are shown in FIG. 10, adhesive surfaces are also provided on two faces that are blind spots. This allows four adhesive surfaces 160a, 160b to be provided on one wiping jig. Note that the wiping jig may have a triangular cross section perpendicular to the extension direction of the rod-shaped member, with adhesive surfaces provided on three faces, or a pentagonal cross section, with adhesive surfaces provided on five faces.

[0051] As shown in wiping jigs 14a and 14b, by providing an attachment surface on each of multiple surfaces of a support member, multiple samples can be obtained with one wiping jig 14a, 14b. The method of using wiping jigs 14a and 14b is not limited, and test objects from different objects may be attached to each attachment surface, or test objects obtained from the same object may be attached to multiple attachment surfaces.

[0052] Fig. 11 is a schematic diagram showing an example of a wiping jig. The wiping jig 14c shown in Fig. 11 has a structure in which the attachment surface is detachable from the support member. The wiping jig 14c has a support member 170 and a flexible member 172. The attachment surface 40 is formed on the surface of the flexible member 172. The flexible member 172 is a deformable rigid member. The support member 170 has a plurality of holes 174 formed therein.

[0053] The wiping jig 14c is wrapped around the support member 170 with the attachment surface 40 facing one surface of the flexible member 172, and a portion of the flexible member 172 is inserted into the hole 174, thereby fixing the flexible member 172 to the support member 170. This allows the attachment surface 40 to be exposed on the surface of the support member 170. Furthermore, the flexible member 172 can be removed from the support member 170 by removing the flexible member 172 from the hole 174.

[0054] The wiping jig 14c allows the attachment surface 40 to be detachable from the support member 170, thereby making it possible to reuse the support member 170. Also, only the flexible member 172 can be the subject of analysis.

[0055] 11, the attachment surface of the wiping jig 14 is detachable from the support member, but the method for forming the attachment surface on the support member is not particularly limited. The attachment surface of the wiping jig may be fixed to the surface of the support member by chemical bonding, for example, bonding with an adhesive.

[0056] It is preferable that the reaction vessel 16 be capable of holding a plurality of wiping jigs 14. Fig. 12 is a schematic diagram showing an example of a reaction vessel. The reaction vessel 16a shown in Fig. 12 has a support 170 with a pentagonal cross section. Furthermore, the wiping jig 14 has a magnet 172 disposed on the surface opposite the attachment surface 40. The reaction vessel 16a can fix the wiping jig 14 to each surface by bringing the magnet 172 of the wiping jig 14 into contact with each of the pentagonal surfaces of the support 170.

[0057] Fig. 13 is a schematic diagram showing an example of a reaction vessel. Reaction vessel 16b shown in Fig. 13 has a support 180 with a pentagonal cross section. Reaction vessel 16a has fixing jigs 182 arranged on each of the pentagonal faces of support 180. Fixing jig 182 is a claw-shaped member that holds the end face of support member 42 of wiping jig 14. Fixing jig 182 supports wiping jig 14.

[0058] As shown in Figures 12 and 13, the structure allows multiple wiping jigs 14 to be processed simultaneously, which makes it possible to improve work efficiency. Furthermore, even when using liquids with a short usable period, the ability to process multiple wiping jigs at once reduces the amount of unused liquid that is discarded. Furthermore, by using a structure in which the wiping jigs are held in place by magnets or a mechanical structure, as in this embodiment, the wiping jigs can be fixed to the supports 170 and 180 without using substances that may dissolve in the liquid, such as adhesives.

[0059] Fig. 14 is a schematic diagram showing the general configuration of a microorganism detection device according to another embodiment. Detailed description of the configuration of the microorganism detection device 10b shown in Fig. 14 that is similar to that of the microorganism detection device 10 will be omitted, and only points unique to the microorganism detection device 10b will be described.

[0060] The microorganism detection device 10b includes a test target acquisition unit 12, a wiping tool 14, a reaction vessel 16, a blocking processing unit 18, a primary antibody liquid supply unit 20, a secondary antibody liquid supply unit 24, a discharge unit 26, a detection unit 28, a control unit 30, and a measurement unit 300. The measurement unit 300 has a sampling line 304 and a measurement device 306. The sampling line 304 acquires a portion of the liquid in the reaction vessel 16 and supplies it to the measurement device 306. The sampling line 304 returns the liquid measured by the measurement device 306 to the reaction vessel 16. The liquid measured by the measurement device 306 may be discarded.

[0061] The measuring device 306 measures the protein concentration contained in the liquid (sample liquid) supplied from the sampling line 304 to detect the state inside the reaction vessel 16. The measuring device 306 measures the sample liquid using the BCA method, Bradford method, UV method, etc. The liquids to be measured by the measuring device 306 are liquids subjected to blocking treatment, primary antibody treatment, and secondary antibody treatment. The measuring unit 300 adds Coomassie brilliant blue, BCA (bicinchoninic acid), copper sulfate (Cu 2+ ) or a buffer solution is supplied and stirred, and the resulting test substance is measured using a spectrophotometer.

[0062] Here, the BCA method uses BCA (bicinchoninic acid) and copper sulfate (Cu 2+ This method utilizes a colorimetric reaction between copper ions (Cu ions) and proteins. Under alkaline conditions, the peptide bonds in proteins reduce divalent copper ions to monovalent copper ions, which are then chelated by BCA to form a stable complex that absorbs violet light, turning the solution from green to purple. The reduction reaction is proportional to the protein concentration, and the protein concentration can be calculated using the absorbance of the solution at 562 nm (purple) using a plate reader or spectrophotometer. This method can be used for protein concentrations ranging from 20 μg / ml to 2000 μg / ml.

[0063] The Bradford assay (Coomassie Brilliant Blue assay) is a method for detecting the reaction of Coomassie Brilliant Blue. Coomassie Brilliant Blue is a triphenylmethane dye that changes color from cationic, unbound red to anionic blue upon binding to proteins. In the presence of proteins, the negative sulfonic acid group of the red dye molecule donates an electron to the positive side chains of lysines and arginines. This destabilizes the protein, promoting van der Waals interactions and exposing hydrophobic residues to the dye, forming a stable complex with the blue anionic dye. The absorption peak of the blue form is at 595 nm, and the optical density of this complex has a linear relationship with protein content within a certain range. This assay can be used for protein concentrations ranging from 10 ng / ml to 2000 ng / ml.

[0064] The UV method uses an ultraviolet spectrophotometer to measure protein concentration. It takes advantage of the fact that aromatic amino acids (tryptophan, lysine, phenylalanine, etc.) in protein molecules contain conjugated double bonds that can absorb ultraviolet light. The maximum absorption peak is at a wavelength of 280 nm. Based on the Beer-Lambert law, the concentration can be calculated using ultraviolet absorption. It can be used when the protein concentration ranges from 50 μg / ml to 2000 μg / ml.

[0065] Here, the microbial detection device 10b has a blocking agent concentration of, for example, 10 mg / ml or less, a primary antibody usage concentration of 0.2 μg / ml or more and 10 μg / ml or less, and a secondary antibody usage concentration of 50 ng / ml or more and 1000 ng / ml or less.

[0066] 15 is a flowchart showing an example of processing by the microorganism detecting device 10b. The microorganism detecting device 10b performs this processing in at least one of the blocking processing, the primary antibody processing, and the secondary antibody processing.

[0067] The microorganism detecting device 10b collects the sample liquid (step S52). The microorganism detecting device 10b obtains a part of the liquid in the reaction vessel 16 as the sample liquid.

[0068] The microorganism detecting device 10b measures the protein concentration (step S54). The microorganism detecting device 10b processes the sample liquid with the measuring device 306 and measures the protein concentration of the sample liquid. The method to be used can be switched depending on the target concentration of the sample liquid to be measured.

[0069] Next, the microorganism detecting device 10b determines whether the protein concentration is equal to or greater than a threshold concentration (step S56). The threshold concentration is set for each of the blocking treatment, primary antibody treatment, and secondary antibody treatment. If the microorganism detecting device 10b determines that the protein concentration is not equal to or greater than the threshold concentration (No in step S56), it returns to step S52 and performs measurement. If the microorganism detecting device 10b determines that the protein concentration is equal to or greater than the threshold concentration (Yes in step S56), it determines whether a predetermined time or more has passed (step S58). In other words, it determines whether the state of the threshold concentration has continued for a predetermined time or more. The predetermined time is also set for each of the blocking treatment, primary antibody treatment, and secondary antibody treatment.

[0070] If the microorganism detection device 10b determines that the predetermined time or more has not passed (No in step S58), it returns to step S52 and performs measurement. If the microorganism detection device 10b determines that the predetermined time or more has passed (Yes in step S58), it ends the processing process (step S60) and ends the determination process in FIG.

[0071] The microorganism detecting device 10b can automatically switch between processes by having the control unit 30 determine whether to end each process based on the protein concentration measured by the measurement unit 300.

[0072] The present disclosure discloses the following inventions, but is not limited to the following. (1) a reaction vessel that holds an adhesion surface to which an object to be inspected is attached; a blocking treatment unit that performs a blocking treatment on the attachment surface of the reaction vessel; a primary antibody solution supply unit that supplies a primary antibody solution containing a primary antibody that captures a protein to the reaction vessel; a secondary antibody solution supply unit that supplies a secondary antibody solution containing a secondary antibody that reacts with the primary antibody to increase the reaction during measurement to the reaction vessel; a cleaning liquid supply unit that supplies a cleaning liquid to the reaction vessel; a discharge part for discharging liquid from the reaction vessel; a detection unit that measures the test object treated in the reaction vessel and detects microorganisms contained in the test object; a control unit that performs a blocking process on the adhesion surface in the reaction vessel, and then supplies and discharges a primary antibody liquid and a secondary antibody liquid in sequence into the reaction vessel.

[0073] (2) The microbial detection device according to (1), wherein the blocking processing unit has a blocking liquid supply unit that supplies a blocking liquid.

[0074] (3) The microorganism detection device according to (1) or (2), wherein the reaction vessel has a stirring means for stirring the liquid therein.

[0075] (4) A measuring unit for measuring the protein concentration in the reaction vessel; The microorganism detection device according to any one of (1) to (3), wherein the control unit determines completion of processing of each step based on the measurement result of the measurement unit.

[0076] (5) a wiping tool having the adhesion surface and removably held in the reaction vessel; The wiping jig includes a support member that supports the adhesion surface; The microorganism detection device according to any one of (1) to (4), further comprising: a rod-shaped member connected to the support member and held by an operator.

[0077] (6) The microbial detection device according to (5), wherein the wiping tool has the attachment surface on a plurality of surfaces of the support member.

[0078] (7) The reaction vessel has a plurality of holding mechanisms for holding the wiping jig, The microorganism detection device according to (5) or (6), wherein the holding mechanism holds the support member by magnetic force.

[0079] (8) The microbial detection device according to any one of (5) to (7), wherein the adhering surface of the wiping tool is fixed to the support member by chemical adhesion.

[0080] (9) The microbial detection device according to any one of (5) to (7), wherein a part of the attachment surface of the wiping jig is inserted into a hole in the support member and fixed to the support member.

[0081] (10) The microbial detection device according to any one of (1) to (9), wherein the blocking processing unit is disposed in the reaction vessel and has a drying unit that dries the attachment surface.

[0082] (11) performing a blocking process on the adhesion surface of the test object held in the reaction vessel and having the test object adhered thereto; Discharging the washing solution and supplying a primary antibody solution containing a primary antibody that captures proteins into the reaction vessel; Discharging the primary antibody solution and supplying a washing solution; Discharging the washing solution and supplying a secondary antibody solution containing a secondary antibody that reacts with the primary antibody and increases the reaction during measurement to the reaction vessel; Discharging the secondary antibody solution and supplying a washing solution; measuring the test object treated in the reaction vessel and detecting microorganisms contained in the test object; A microbial detection method that automatically controls the supply and discharge of liquids in each section, and sequentially supplies and discharges blocking liquid, primary antibody liquid, and secondary antibody liquid into the reaction vessel. [Explanation of symbols]

[0083] 10 Microbial detection device 12 Inspection target acquisition unit 14 Wiping jig 16 Reaction vessel 18 Blocking processing section (blocking solution supply section) 20 Primary antibody solution supply section 22 Secondary antibody solution supply section 24 Cleaning liquid supply unit 26 Discharge section 28 Detector 30 Control Unit 40 Adhesion surface 42 Support member 44 Rod-shaped member 50 Blocking solution tank 52, 62, 72, 82 supply route 54, 64, 74, 84, 96, 98 On-off valves 60 Primary antibody solution tank 70 Secondary antibody solution tank 80 cleaning solution tank 90 Exhaust channel 92 Drainage tank 94 Recovery Route

Claims

1. a reaction vessel that holds an adhesion surface to which an object to be inspected is attached; a blocking treatment unit that performs a blocking treatment on the attachment surface of the reaction vessel; a primary antibody solution supply unit that supplies a primary antibody solution containing a primary antibody that captures a protein to the reaction vessel; a secondary antibody solution supply unit that supplies a secondary antibody solution containing a secondary antibody that reacts with the primary antibody to increase the reaction during measurement to the reaction vessel; a cleaning liquid supply unit that supplies a cleaning liquid to the reaction vessel; a discharge part for discharging liquid from the reaction vessel; a detection unit that measures the test object treated in the reaction vessel and detects microorganisms contained in the test object; a control unit that executes a process of supplying a primary antibody solution and a secondary antibody solution to the reaction vessel in order and then discharging them after performing a blocking process on the adhesion surface in the reaction vessel; a wiping tool having the adhesion surface and removably held in the reaction vessel; The wiping jig includes a support member that supports the adhesion surface; a rod-shaped member connected to the support member and held by an operator.

2. The microorganism detection device according to claim 1 , wherein the blocking processing unit includes a blocking liquid supply unit that supplies a blocking liquid.

3. 2. The microorganism detection device according to claim 1, wherein the reaction vessel has a stirring means for stirring the liquid therein.

4. a measuring unit that measures the protein concentration in the reaction vessel; The microorganism detection device according to claim 1 , wherein the control unit determines the end of processing in each step based on the measurement result of the measurement unit.

5. The microbial detection device according to claim 1 , wherein the wiping tool has the attachment surface on a plurality of surfaces of the support member.

6. the reaction vessel has a plurality of holding mechanisms for holding the wiping jigs; 2. The microorganism detection device according to claim 1, wherein the holding mechanism holds the support member by magnetic force.

7. The microbial detection device according to claim 1 , wherein the adhering surface of the wiping tool is fixed to the support member by chemical adhesion.

8. The microorganism detection device according to claim 1 , wherein the wiping jig is fixed to the support member by inserting a part of the adhesion surface into a hole in the support member.

9. The microorganism detection device according to claim 1 , wherein the blocking processing unit has a drying unit disposed in the reaction vessel and configured to dry the adhesion surface.

10. A step of attaching an inspection object to an attachment surface of a wiping jig; a step of holding the wiping jig in a reaction vessel and performing a blocking process on the adhesion surface on which the inspection object is adhered; supplying a primary antibody solution containing a primary antibody that captures a protein into the reaction vessel; Discharging the primary antibody solution and supplying a washing solution; Discharging the washing solution and supplying a secondary antibody solution containing a secondary antibody that reacts with the primary antibody and increases the reaction during measurement to the reaction vessel; Discharging the secondary antibody solution and supplying a washing solution; measuring the test object treated in the reaction vessel and detecting microorganisms contained in the test object; The system controls the supply and discharge of liquids in each section, and automatically supplies and discharges blocking solution, primary antibody solution, and secondary antibody solution to the reaction vessel in that order. Microbial detection methods.

11. The wiping jig comprises: a support member that supports the adhesion surface; The microorganism detection method according to claim 10, further comprising a rod-shaped member connected to the support member and held by an operator.

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