WO2019167539A1

WO2019167539A1 - Method for confirming state of immunoassay device, and immunoassay device

Info

Publication number: WO2019167539A1
Application number: PCT/JP2019/003455
Authority: WO
Inventors: 卓弥能田; 徹植村; 金子　周平
Original assignee: シスメックス株式会社
Priority date: 2018-02-28
Filing date: 2019-01-31
Publication date: 2019-09-06
Also published as: JP2019148558A; JP7161851B2

Abstract

The objective of the present invention is to enable the state of a function for carrying out immunoconjugate transfer in an immunoassay device to be confirmed in a simple manner. This method for confirming the state of an immunoassay device (100) is a method for confirming the state of the immunoassay device (100) which performs an immunoconjugate transfer process in which an immunoconjugate (84), supported on the solid phase carrier (22) and including a substance (81) being tested for in a sample and a labeling substance (21), is liberated from the solid phase carrier (22) in a first container (11) accommodating the immunoconjugate (84), and a liquid phase in the first container (11) is transferred to a second container (12), wherein the method includes: a step of transferring the liquid phase in the first container (11) accommodating a control sample (20) containing at least the labeling substance (21) to the second container (12); a step of detecting the labeling substance (21) in the second container (12) to which the liquid phase has been transferred; and a step of assessing the state of the immunoassay device (100) which performs the immunoconjugate transfer process, on the basis of the detection result of the labeling substance (21) in the second container (12).

Description

Method for confirming state of immunoassay device and immunoassay device

This invention relates to an immunoassay device for measuring a test substance in a specimen.

As a technique for increasing the sensitivity of immunoassays, there is an immunoassay apparatus using an immune complex transfer method (see, for example, Patent Document 1).

As shown in FIG. 23, the immunoassay device disclosed in Patent Document 1 forms an immune complex 913 including a test substance 911 and a labeling substance 912 on a solid phase carrier 914 in a first container 901. Then, the immune complex 913 is released from the solid phase carrier 914 by the release reagent 915. Then, the immunoassay device performs the immune complex transfer method in which the liquid phase containing the released immune complex 913 is transferred to the second container 902 while leaving the solid phase carrier 914 in the first container 901, The signal based on the labeling substance 912 contained in the immune complex 913 in the two containers 902 is detected.

JP 2017-49059 A

By the way, since the measurement result obtained by the immunoassay device becomes a judgment material for diagnosing a disease or determining a treatment policy, the reliability of the measurement result is required. As a method for ensuring the reliability of measurement results, there is a conventional method of measuring a quality control sample containing a known concentration of a test substance, which can be carried out easily. Used in measuring equipment.

However, in the immunoassay device using the immune complex transfer method, functions that are not available in the conventional immunoassay device are added to execute the immune complex transfer method. For this reason, in the conventional method of measuring a quality control sample, there is a possibility that it is not possible to check the normality or abnormality of the function for executing the immune complex transfer method. Even in the above-mentioned Patent Document 1 using the immune complex transfer method, a method for confirming the state of the function for executing the immune complex transfer method is not disclosed, and the function for executing the immune complex transfer method is not disclosed. It is desirable to be able to easily check the state of the above.

The present invention is directed to making it possible to easily check the status of the function for executing the immune complex transfer method in the immunoassay device.

The method for confirming the state of an immunoassay device according to the first aspect of the present invention comprises an immune complex (84) comprising a test substance (81) and a labeling substance (21) in a specimen and supported on a solid phase carrier (22). ) Is released from the solid phase carrier (22) and the liquid phase in the first container (11) is transferred to the second container (12). A method for confirming a state of an immunoassay device (100) that performs body transfer treatment, wherein a liquid phase in a first container (11) containing a control sample (20) containing at least a labeling substance (21) is contained in a second container. (12), the step of detecting the labeling substance (21) in the second container (12) to which the liquid phase has been transferred, and the detection result of the labeling substance (21) in the second container (12). And a step of determining the state of the immunoassay device (100) performing the complex transfer process based on

In the method for confirming the state of the immunoassay device according to the first aspect, a complex is formed with respect to the first container (11) containing the control sample (20) containing the labeling substance (21) by being configured as described above. The labeling substance (21) in the control sample (20) transferred to the second container (12) can be detected by performing a liquid phase transfer operation similar to the transfer process. If the complex transfer process is not performed normally, an abnormal detection result deviating from the expected detection result is acquired. Thereby, based on the detection result of the labeling substance (21) in the second container (12), it is possible to determine the state of the immunoassay device, such as whether or not the complex transfer process has been successfully performed. As a result, it is possible to easily check the status of the function for executing the immune complex transfer method in the immunoassay device.

In the method for confirming the state of the immunoassay device according to the first aspect, preferably further comprising the step of determining an abnormality in the complex transfer process based on the detection result of the labeling substance (21) in the second container (12). Prepare. With this configuration, for example, whether or not there is an abnormality in the complex transfer process by determining the detection result in the immunoassay device (100) that has detected the labeling substance (21) in the control sample (20). Can be determined.

In this case, preferably, in the step of determining abnormality in the complex transfer treatment, the function of collecting the solid phase carrier (22) in the first container (11) and the liquid phase in the first container (11) are set to the second. Determining at least one abnormality of the function of transferring to the container (12). Here, in the complex transfer treatment, in order to move the liquid phase containing the immune complex (84) to the second container (12) and leave the solid phase carrier (22) in the first container (11), Whether the function of collecting the solid phase carrier (22) in the first container (11) is normal is important. In addition, the function of transferring the liquid phase in the first container (11) to the second container (12) in order to detect the immune complex (84) transferred to the second container (12) by the complex transfer treatment. Whether or not is normal is important. According to the above configuration, since it is possible to determine the presence / absence of at least one of these functions, it is particularly useful for ensuring the reliability of the measurement result.

In the case of including the step of determining abnormality in the complex transfer treatment, preferably, the detected value of the labeling substance (21) in the second container (12) exceeds the reference value (V1), or the detected value is the reference value. Abnormality is determined based on falling below (V2). If comprised in this way, based on the detection value obtained by the measurement in a normal state, the reference value (V1, V2) for distinguishing between normal and abnormality is preset, and a detection value and a reference | standard By simply comparing the values (V1, V2), the abnormality determination of the complex transfer process can be easily performed.

In the method for confirming the state of the immunoassay device according to the first aspect, preferably, before the step of transferring the liquid phase in the first container (11) to the second container (12), by the immunoassay device (100), The method further includes the step of dispensing the control sample (20) into the first container (11). If comprised in this way, even if the user of the immunoassay apparatus (100) and the service staff who performs maintenance service of the apparatus do not prepare the management sample (20) in the first container (11) in advance, the immunoassay The management sample (20) can be easily accommodated in the first container (11) by the apparatus (100).

In the method for confirming the state of the immunoassay device according to the first aspect, preferably, the management sample (20) includes a solid phase carrier (22) to which a labeling substance (21) is bound, and is contained in the first container (11). In the step of transferring the liquid phase to the second container (12), the solid phase carrier (22) to which the labeling substance (21) is bound is collected in the first container (11). With this configuration, the solid phase carrier (22) to which the labeling substance (21) is bound is collected in the first container (11). For example, when the detection value of the labeling substance (21) increases, When the phase is transferred to the second container (12), it is determined that the solid support (22) is not sufficiently collected and the solid support (22) is also likely to be transferred to the second container (12). it can. Therefore, it can be easily confirmed whether or not the function of collecting the solid phase carrier (22) in the first container (11) in the complex transfer treatment is normal.

In this case, preferably, the solid phase carrier (22) is a magnetic particle, and the process of collecting the solid phase carrier (22) includes a process of collecting the magnetic particles by the magnetic force source (52) and includes the second container (12). ) Further includes a step of determining abnormality of the magnetic flux collecting function by the magnetic force source (52) based on the detection result of the labeling substance (21) in the parenthesis. In this specification, the “magnetic collecting function” is a function of collecting magnetic particles, and that the normal magnetic collecting function means that the first container (11) to the second container (12 ) Indicates that the carry-over amount of the magnetic particles transferred together with the liquid phase is suppressed within an allowable range. According to this configuration, the magnetic particles to which the labeling substance (21) is bonded are collected, so that the magnetic source (52) collects the magnetic particles based on the detection value of the labeling substance (21) in the second container (12). It is possible to easily determine whether the magnetic function is normal or abnormal.

When determining the abnormality of the magnetic flux collecting function by the magnetic source (52), preferably, when the detected value of the labeling substance (21) exceeds the reference value (V1), the abnormal magnetic flux collecting function by the magnetic source (52) is detected. Judge that there is. If comprised in this way, when the detection value of a labeled | labeling substance (21) exceeds reference value (V1), when transferring a liquid phase to a 2nd container (12), it has not fully collected the magnetic particle. Since the possibility is high, the magnetic flux collecting function by the magnetic source (52) can be easily determined by comparing the detected value with the reference value (V1).

In this case, preferably, the reference value (V1) is an expected detection value of the labeling substance (21) corresponding to the allowable upper limit amount of the magnetic particles transferred to the second container (12) without being collected. If comprised in this way, when the magnetic particle of the quantity exceeding an allowable upper limit amount is moved to the 2nd container (12), it can determine easily that the magnetism collection function by a magnetic source (52) is abnormal. . Therefore, the reliability of the measurement result can be ensured by confirming that the allowable upper limit amount is not exceeded.

In the configuration in which the solid phase carrier is collected, preferably, the known concentration in the first container (11) is set before the step of transferring the liquid phase in the first container (11) to the second container (12). A sample containing the test substance (81), a labeling substance (83) binding to the test substance (81), a capture substance (86) binding to the test substance (81), and binding to the capture substance (86) The method further includes a step of contacting the solid phase carrier (82). If comprised in this way, the solid-phase carrier (82) which the label | marker substance (83) couple | bonded will be the 1st container (11) by the process process similar to the case where the test substance containing a test substance (81) is actually measured. Can be formed inside.

In this case, preferably, the control sample (20) includes the first solid phase carrier (82a) to be dispensed into the first container (11) before the complex transfer process when the specimen is measured. If comprised in this way, the solid-phase carrier which the label | marker substance (83) couple | bonded using the 1st solid-phase carrier (82a) dispensed when actually measuring the test substance containing a test substance (81) will be carried out. (82) can be formed in the first container (11). That is, since it is not necessary to separately prepare a dedicated solid phase carrier (22) for confirming the state of the immunoassay device (100), the convenience of the immunoassay device (100) can be improved.

When the sample including the test substance (81), the labeling substance (83), the capture substance (86), and the solid phase carrier (82) are provided, preferably, the control sample (20) A second solid phase carrier (82b) to be dispensed into the second container (12) after the complex transfer treatment at the time of measurement is included. If comprised in this way, the solid-phase carrier which the label | marker substance (83) couple | bonded using the 2nd solid-phase carrier (82b) dispensed when actually measuring the test substance containing a test substance (81) will be carried out. (82) can be formed in the first container (11). That is, since it is not necessary to separately prepare a dedicated solid phase carrier (22) for confirming the state of the immunoassay device (100), the convenience of the immunoassay device (100) can be improved.

When the sample including the test substance (81), the labeling substance (83), the capture substance (86), and the solid phase carrier (82) are brought into contact with each other, the liquid in the first container (11) is preferably used. Prior to the step of transferring the phase to the second container (12), the solid phase carrier (82) bound to the immune complex (84) containing the test substance (81), the labeling substance (83), and the capture substance (86). And a step of performing BF separation for separating the liquid phase. If comprised in this way, the labeling substance (83) in the liquid phase which did not couple | bond with the solid-phase carrier (82) can be excluded from a 1st container (11) by BF separation. Therefore, in the step of transferring the liquid phase in the first container (11) to the second container (12), the mixing of the labeling substance (83) in the liquid phase can be suppressed, so that the state of the immunoassay device (100) is confirmed. The detection accuracy of the labeling substance (83) for the purpose can be improved.

In this case, preferably, in the complex transfer treatment, the immune complex (84) is released from the solid phase carrier (82) by dispensing the free reagent (85) into the first container (11), and BF After the step of separating, before the step of transferring the liquid phase in the first container (11) to the second container (12), the liquid phase not containing the free reagent (85) is separated into the first container (11). The process of adding is further provided. If comprised in this way, it replaces with the free reagent (85) dispensed when actually measuring the test substance containing a test substance (81), and the liquid phase (25) which does not contain a free reagent (85) is used. Since dispensing is performed, the state of the immunoassay device (100) can be confirmed by the same operation as dispensing the free reagent (85) during actual sample measurement. Even in this case, since the binding between the labeling substance (83) and the solid phase carrier (82) is not eliminated, the function of collecting the solid phase carrier (82) can be appropriately confirmed.

When the control sample (20) includes the solid phase carrier (22) to which the labeling substance (21) is bound, preferably, the control sample (20) is not bound to the test substance (81) and the labeling substance ( 21) and a third solid phase carrier (23). If comprised in this way, using the 3rd solid-phase carrier (23) which does not contain the to-be-tested substance (81) as a reagent for exclusive use for the state confirmation of an immunoassay device (100), a solid-phase carrier (22 ) Can be confirmed. When the solid phase carrier (22) binds not only to the labeling substance (21) but also to the test substance (81), the solid phase carrier (22) and the labeling substance (which can be bound according to the type of the test substance (81)) 21), a plurality of types are prepared, whereas in the third solid phase carrier (23) that does not include the test substance (81), the immunoassay apparatus (100) is not dependent on the type of the test substance (81). The status can be checked. Therefore, it is possible to more easily check the state of the immunoassay device (100).

In the method for confirming the state of the immunoassay device according to the first aspect, preferably, the control sample (20) includes a solution in which the labeling substance (21) is dissolved. If comprised in this way, since the labeling substance (21) exists as a liquid phase in the 1st container (11), for example, when the detection value of the labeling substance (21) does not rise, the liquid phase is set in the second container. When moving to (12), it can be determined that there is a high possibility that the liquid phase has not been properly transferred from the first container (11). Therefore, it can be easily confirmed whether or not the function of transferring the liquid phase in the first container (11) to the second container (12) in the complex transfer treatment is normal.

In this case, preferably, in the step of transferring the liquid phase in the first container (11) to the second container (12), the liquid phase in the first container (11) is sucked and the sucked liquid phase is sucked into the second container. (12) includes a dispensing process to be discharged, and further includes a step of determining abnormality of the dispensing function based on the detection result of the labeling substance (21) in the second container (12). In the present specification, the “dispensing function” includes a function of sucking the liquid phase in the first container (11) and a function of discharging the sucked liquid phase to the second container (12). The normal function means that the liquid phase can be sucked from the first container (11) and the sucked liquid phase can be discharged to the second container (12). If comprised in this way, it can be easily determined whether the dispensing function which performs suction | inhalation of a liquid phase and discharge of a liquid phase is normal based on the detection value of a labeling substance (21).

When determining an abnormality in the dispensing function, preferably, it is determined that there is an abnormality in the dispensing function when the detected value of the labeling substance (21) is lower than the reference value (V2). With this configuration, when the detected value of the labeling substance (21) is lower than the reference value (V2), the liquid containing the labeling substance (21) from the first container (11) to the second container (12). Since there is a high possibility that the phases have not been appropriately transferred, the dispensing function can be easily determined by comparing the detected value with the reference value (V2).

In this case, preferably, the reference value (V2) is a value set as an allowable limit of an expected detection value of the labeling substance (21) contained in the control sample (20). If comprised in this way, even if the process which moves the liquid phase containing a labeling substance (21) to a 2nd container (12) is performed, when a detected value is less than a reference value (V2), a dispensing function is abnormal. It can be easily determined. Therefore, the reliability of the measurement result can be ensured by confirming that the detected value exceeds the reference value (V2).

When the management sample includes a solution in which the labeling substance (21) is dissolved, the management sample (20) preferably includes the labeling substance (21) to be dispensed into the first container (11) at the time of measurement of the specimen. . If comprised in this way, the liquid phase in a 1st container (11) will be made into a 2nd container using the label | marker substance (21) dispensed when actually measuring the test substance containing a test substance (81). It can be confirmed whether or not the function transferred to (12) is normal. That is, since it is not necessary to separately prepare a dedicated labeling substance (21) for confirming the state of the immunoassay device (100), the convenience of the immunoassay device (100) can be improved.

In the method for confirming the state of the immunoassay device according to the first aspect, the labeling substance (21) contained in the control sample (20) is an enzyme, and the labeling substance (21) in the second container (12) is detected. May be performed by adding a substrate of the enzyme to the second container (12) and measuring a signal generated from a reaction product generated by the enzyme reaction.

In the method for confirming the state of the immunoassay device according to the first aspect, in the first container (11), a sample containing a test substance (81) having a known concentration or a sample not containing the test substance (81), a test Contacting a labeling substance (83) that binds to the substance (81), a capture substance (86) that binds to the test substance (81), and a solid phase carrier (82) that binds to the capture substance (86); After dispensing a free reagent (85) for releasing the immune complex (84) containing the labeling substance (83) and the capture substance (86) from the solid phase carrier (82) in one container (11), the first You may further provide the process of moving the liquid phase in a container (11) to a 2nd container (12), and the process of detecting the labeling substance (83) which exists in a 2nd container (12). That is, an accuracy control method for measuring a control sample in the second container (12) by performing an operation similar to that for measuring a specimen using a control sample having a known concentration or a sample not containing a test substance. Further implementation may be performed.

The immunoassay device (100) according to the second aspect of the present invention comprises an immune complex (84) comprising a test substance (81) and a labeling substance (21) in a specimen and carried on a solid phase carrier (22). In the first container (11) containing the immunity, the immune complex (84) is released from the solid phase carrier (22), and the liquid phase in the first container (11) is transferred to the second container (12). An immunoassay device (100) for performing a transfer process, wherein a liquid phase in a first container (11) containing a control sample (20) containing at least a labeling substance (21) is transferred to a second container (12). Based on the detection result of the mechanism part (50) which performs processing, the detection part (60) which detects the labeling substance (21) in the second container (12) to which the liquid phase has been transferred, and the detection part (60) And a determination unit (70) for determining an abnormality in the complex transfer process.

The immunoassay device (100) according to the second aspect is configured as described above, whereby complex transfer is performed with respect to the first container (11) containing the control sample (20) containing the labeling substance (21). The labeling substance (21) in the control sample (20) transferred to the second container (12) can be detected by performing a liquid phase transfer operation similar to the processing. If the complex transfer process is not performed normally, an abnormal detection result deviating from the expected detection result is acquired. Thus, the determination unit (70) can determine whether or not the complex transfer process has been successfully performed from the detection result of the labeling substance (21) in the second container (12). The status of the function for carrying out the immune complex transfer method in can be easily confirmed.

In the immunoassay device (100) according to the second aspect, the determination unit (70) preferably has a function and a mechanism for collecting the solid phase carrier (22) in the first container (11) by the mechanism unit (50). An abnormality of at least one of the function of transferring the liquid phase in the first container (11) to the second container (12) by the unit (50) is determined. With this configuration, the determination unit (70) causes at least one of the abnormality of the function of collecting the solid phase carrier (22) in the complex transfer process and the function of transferring the liquid phase to the second container (12). Since the presence or absence can be determined, it is particularly useful for ensuring the reliability of the measurement result.

In this case, preferably, the control sample (20) includes magnetic particles to which the labeling substance (21) is bound, and the mechanism part (50) is a magnetic material to which the labeling substance (21) is bound in the first container (11). A magnetic source (52) for collecting particles is included, and the determination unit (70) determines abnormality of the magnetic collection function by the magnetic source (52). With this configuration, the solid phase carrier (22) to which the labeling substance (21) is bound is collected in the first container (11). For example, when the detection value of the labeling substance (21) increases. There is a possibility that when the liquid phase is transferred to the second container (12), the solid phase carrier (22) is not sufficiently magnetized and the solid phase carrier (22) is also transferred to the second container (12). Can be determined to be high. Therefore, it can be easily determined whether the magnetic flux collecting function by the magnetic force source (52) in the complex transition process is normal or abnormal.

In the configuration in which the determination unit (70) determines an abnormality in at least one of the function of collecting the solid phase carrier (22) and the function of transferring the liquid phase to the second container (12), preferably the control sample ( 20) includes a solution in which the labeling substance (21) is dissolved. The mechanism (50) sucks the liquid phase in the first container (11) and discharges the sucked liquid phase to the second container (12). The determination unit (70) includes a suction pipe (51) that performs the determination, and determines an abnormality in the dispensing function of the suction pipe (51). If comprised in this way, since the labeling substance (21) exists as a liquid phase in the 1st container (11), for example, when the detection value of the labeling substance (21) does not rise, the liquid phase is set in the second container. When dispensing into (12), it can be determined that there is a high possibility that the dispenser by the suction pipe (51) that performs the suction of the liquid phase and the discharge of the liquid phase is not normal. Therefore, the presence or absence of abnormality in the dispensing function by the suction tube (51) in the complex transfer process can be easily confirmed.

In the immunoassay device (100) according to the second aspect, preferably, the mechanism unit (50) includes a sample dispensing unit (120) for dispensing a sample containing the test substance (81), and a labeling substance. A reagent dispensing unit (130) for dispensing a labeling reagent containing (21, 83) and a solid phase reagent containing a solid phase carrier (22, 82), and in the first container (11) The control sample (20) is prepared. If comprised in this way, even if the user and service staff of an immunoassay device (100) do not prepare the management sample (20) in the 1st container (11) in advance, it is easy by the immunoassay device (100). The management sample (20) can be accommodated in the first container (11).

In this case, it is preferable that the reagent dispensing unit (130) removes the free reagent (85) that liberates the immune complex (84) from the solid phase carrier (82) when the sample is measured. 11), when determining an abnormality in the process of transferring the liquid phase in the first container (11) to the second container (12), the free reagent (85) is replaced with the free reagent (85). Dispense the free liquid phase (25). If comprised in this way, it replaces with the free reagent (85) dispensed when actually measuring the test substance containing a test substance (81), and the liquid phase (25) which does not contain a free reagent (85) is used. Since dispensing is performed, the state of the immunoassay device (100) can be confirmed by the same operation as dispensing the free reagent (85) during actual sample measurement. Even in this case, since the binding between the labeling substance (83) and the solid phase carrier (82) is not eliminated, the magnetic flux collecting function of the solid phase carrier (82) can be appropriately confirmed.

In the configuration in which the mechanism part (50) prepares the control sample (20) in the first container (11), the mechanism part (50) preferably includes a test substance (81) and a labeling substance (83). It includes a solid phase carrier (82) bound to the immune complex (84) and a BF separation unit (170) for separating the liquid phase, and the BF separation unit (170) is a control sample in the first container (11). BF separation for (20) is performed. If comprised in this way, the labeling substance (83) in the liquid phase which was not couple | bonded with a solid-phase carrier (82) can be excluded from the inside of the 1st container (11) by BF separation. Therefore, in the step of transferring the liquid phase in the first container (11) to the second container (12), the mixing of the labeling substance (83) in the liquid phase can be suppressed, so that the state of the immunoassay device (100) is confirmed. The detection accuracy of the labeling substance (83) for the purpose can be improved.

According to the present invention, it is possible to easily confirm the state of the function for executing the immune complex transfer method in the immunoassay device.

FIG. 1 is a schematic diagram showing a first example of a state confirmation method for an immunoassay device. FIG. 2 is a schematic diagram showing a second example of the state confirmation method of the immunoassay device. FIG. 3 is a schematic diagram showing an outline of the immunoassay device. FIG. 4 is a diagram for explaining an outline of measurement of a sample by the immunoassay device. FIG. 5 is a schematic plan view showing a specific configuration example of the immunoassay device. FIG. 6 is a schematic diagram for explaining the sample dispensing unit. FIG. 7 is a schematic diagram for explaining the reagent dispensing unit. FIG. 8 is a schematic diagram for explaining the BF separation unit. FIG. 9A is a schematic diagram illustrating a configuration example of a mechanism unit for transferring the liquid phase from the first container to the second container. FIG. 9B is a schematic diagram illustrating a configuration example of a mechanism unit for transferring the liquid phase from the first container to the second container. FIG. 9C is a schematic diagram illustrating a configuration example of a mechanism unit for transferring the liquid phase from the first container to the second container. FIG. 10 is a schematic diagram for explaining the detection unit. FIG. 11 is a diagram for explaining a method of determining a magnetic flux collecting function. FIG. 12 is a diagram for explaining the first embodiment for confirming the magnetism collecting function of the immunoassay device. FIG. 13 is a diagram for explaining the second embodiment for confirming the magnetism collecting function of the immunoassay device. FIG. 14 is a diagram for explaining the third embodiment for confirming the magnetism collecting function of the immunoassay device. FIG. 15 is a diagram for explaining Example 1 performed to confirm the effect of the third embodiment. FIG. 16 shows the measurement results of Example 1. FIG. FIG. 17 is a diagram for explaining a method for determining the dispensing function. FIG. 18 is a diagram for explaining the fourth embodiment for confirming the dispensing function of the immunoassay device. FIG. 19 is a diagram for explaining the fifth embodiment for confirming the dispensing function of the immunoassay device. FIG. 20 is a diagram for explaining Example 2 performed to confirm the effect of the fifth embodiment. FIG. 21 is a diagram showing the measurement results of Example 2. FIG. 22 is a flowchart for explaining the operation of the immunoassay device. FIG. 23 is a diagram for explaining the prior art.

Hereinafter, embodiments will be described with reference to the drawings.

[Overview of status check method of immunoassay device]
First, with reference to FIG. 1 and FIG. 2, the outline | summary of the state confirmation method of the immunoassay apparatus by one Embodiment is demonstrated.

The state confirmation method of the immunoassay apparatus 100 is a state confirmation method of the immunoassay apparatus 100 that performs a complex transfer process in which the liquid phase in the first container 11 is transferred to the second container 12.

The immunoassay apparatus 100 measures a test substance in a specimen using an antigen-antibody reaction. The specimen is a biological sample such as blood collected from a living body, for example. The blood may be whole blood, serum, or plasma. The test substance is, for example, an antigen or antibody contained in blood, a protein, a peptide, or the like.

The first container 11 and the second container 12 are, for example, cylindrical containers that are open at the upper end and closed at the bottom at the lower end, and are reaction containers (cuvettes) that can contain liquids such as specimens and reagents. These containers are, for example, disposable resin containers. In this case, the used container can be discarded as it is. The first container 11 and the second container 12 may be containers having the same shape or different shapes.

In this embodiment, the immunoassay apparatus 100 performs a complex transfer process by an immune complex transfer method. In the immunocomplex transfer method (see FIG. 4), a test substance 81 and a labeling substance 83 in a specimen are contained in an immunocomplex (conjugate by antigen-antibody reaction) 84 that is carried on a solid phase carrier 82. In this method, the immune complex 84 is released from the solid phase carrier 82 in one container 11 and the released immune complex 84 is separated from the solid phase carrier 82. The immunoassay apparatus 100 performs the complex transfer process by leaving the solid phase carrier 82 in the first container 11 and transferring the liquid phase in the first container 11 to the second container 12. When the immunoassay apparatus 100 moves the liquid phase in the first container 11 to the second container 12, the immunoassay apparatus 100 prevents the solid phase carrier 82 from being carried over to the second container 12 together with the liquid phase. Collect the solid support 82.

Thereby, unnecessary labeling substance 83 non-specifically bound to the solid phase carrier 82 in the process of forming the immune complex 84 on the solid phase carrier 82 is separated from the immune complex 84 together with the solid phase carrier 82. . As a result, compared to the case where measurement is performed without performing the immune complex transfer method, the noise level can be lowered, so that the baseline of the measurement data can be lowered and the sensitivity of the immunoassay can be increased.

In the present specification, the immune complex may be a labeling substance (an antibody that binds to the test substance or a labeling substance bound to a light source). The immune complex can be a conjugate of a labeling substance and an antibody. The immune complex can be a conjugate of a labeling substance, an antibody, and a test substance.

<Immunoassay device status confirmation method>
The method for confirming the state of the immunoassay apparatus 100 according to the present embodiment confirms whether or not the function for performing the complex transfer process by the immunoassay apparatus 100 is normal.

As shown in FIG. 1 and FIG. 2, the method for confirming the state of the immunoassay device 100 is a step of transferring the liquid phase in the first container 11 containing the control sample 20 containing at least the labeling substance 21 to the second container 12 ( a), a step (b) of detecting the labeling substance 21 in the second container 12 to which the liquid phase has been transferred, and a detection result of the labeling substance 21 in the second container 12, and a complex transfer process is performed. And (c) determining the state of the immunoassay device 100. That is, in the state confirmation method, as the step (a), the liquid phase transfer operation similar to the complex transfer process at the time of sample measurement by the immunoassay apparatus 100 is performed. In step (b), the control sample 20 in the transferred second container 12 is measured. In step (c), the state is determined based on the detection result obtained in step (b).

Step (a) is performed by, for example, a dispensing process in which the liquid phase in the first container 11 is sucked by the suction tube 51 provided in the immunoassay apparatus 100 and the sucked liquid phase is discharged into the second container 12.

In step (b), for example, the labeling substance 21 in the second container 12 is detected by a detection unit provided in the immunoassay apparatus 100.

In step (c), for example, the determination unit provided in the immunoassay apparatus 100 performs state determination based on the detection result. Step (c) can also be performed by, for example, a computer connected to the immunoassay device 100, a server device that can communicate with the immunoassay device 100 via a network, and the like.

<Labeling substance>
The labeling substance is not particularly limited as long as it can emit a detectable or measurable signal. For example, an enzyme, a fluorescent substance, a radioisotope, etc. are mentioned. Examples of the enzyme include alkaline phosphatase, β-galactosidase, peroxidase, glucose oxidase, tyrosinase, acid phosphatase, and luciferase, but are not particularly limited. Fluorescent substances include fluorescein isothiocyanate (FITC), coumarin, rhodamine, fluorescein, Cy3, Cy5, Hoechst 33342, 4 ′, 6-diamidino-2-phenylindole (DAPI), propidium iodide (PI), Alexa Fluor ( Fluorescent dyes such as Molecular Probes (registered trademark) series, fluorescent proteins such as green fluorescent protein (GFP), and the like are exemplified, but not limited thereto. Examples of the radioisotope include 125I, 14C, and 32P, but are not particularly limited.

In addition, in this specification, the labeling substance 21 is a concept including a labeling antibody (an antibody that binds to a test substance and a labeling substance bound) used as a measurement reagent.

In this specification, the labeling substance 21 included in the management sample 20 may be the same labeling substance as the labeling substance 83 (see FIG. 4) included in the immune complex 84 at the time of measurement of the specimen, or different. It may be a labeling substance.

The detection of the labeling substance is not particularly limited as long as it is performed by an appropriate method according to the type of label used for the labeling substance. What is necessary is just to detect the abundance of a labeling substance using the detection means corresponding to the labeling substance. For example, when the label used for the labeling substance is an enzyme, the measurement can be performed by measuring light, color, etc. generated by reacting a substrate with the enzyme. As a detection unit in this case, a photomultiplier tube, a spectrophotometer, a luminometer, or the like can be used. Further, when the labeling substance is a radioisotope, a scintillation counter or the like can be used as the detection unit. When the labeling substance is a fluorescent substance, the labeling substance can be detected using a fluorescence detector capable of detecting the emitted fluorescence.

When the labeling substance is an enzyme, a known substrate may be appropriately selected according to the enzyme used. For example, when alkaline phosphatase is used as the enzyme, the substrate is CDP-Star®, (4-chloro-3- (methoxyspiro {1,2-dioxetane-3,2 ′-(5′-chloro) trixiro [3.3.1.13,7] decan} -4-yl) phenyl phosphate disodium), CSPD® (3- (4-methoxyspiro {1,2-dioxetane-3,2- ( Chemiluminescent substrates such as 5′-chloro) tricyclo [3.3.1.13,7] decan} -4-yl) phenyl phosphate disodium); p-nitrophenyl phosphate, 5-bromo-4-chloro- Luminescent substrates such as 3-indolyl phosphate (BCIP), 4-nitroblue tetrazolium chloride (NBT), iodonitrotetrazolium (INT); 4-methylumberif Fluorescent substrates such as phenyl phosphate (4MUP); 5-bromo-4-chloro-3-indolyl phosphate (BCIP), disodium 5-bromo-6-chloro-indolyl phosphate, p-nitrophenyl phosphorus, etc. The chromogenic substrate can be used.

The detection result reflecting the abundance of the labeling substance 21 is acquired by the detection. The detection result can be acquired in the form of numerical information as a detection value corresponding to the amount of the labeling substance 21 present.

(Management sample including solid phase carrier bound with labeling substance)
As shown in FIG. 1, when the management sample 20 includes a solid phase carrier 22 to which a labeling substance 21 is bound, a process of collecting the solid phase carrier 22 is performed when performing the step (a). By the step (a), the liquid phase is moved from the first container 11 to the second container 12 while leaving the solid phase carrier 22. For this reason, the second container 12 contains a liquid phase that does not include the solid phase carrier 22 to which the labeling substance 21 is bound.

<Solid support>
The solid phase carrier is, for example, a known particle used in immunoassay. Examples of the particles include magnetic particles, latex particles, erythrocytes, and gelatin particles. In order to separate the liquid phase from the first container 11, it is preferable to use magnetic particles as the solid phase carrier 22. The magnetic particle may be any particle that contains a magnetic material as a base material and is used for normal immunoassay. For example, magnetic particles using Fe ₂ O ₃ and / or Fe ₃ O ₄ , cobalt, nickel, phyllite, magnetite, etc. as the substrate can be used.

In the present specification, the solid phase carrier 22 included in the management sample 20 may be the same solid phase carrier 82 as the solid phase carrier 82 that binds to the immune complex 84 when the specimen is measured, or a different solid phase carrier. It may be.

The binding between the labeling substance and the solid phase carrier may be performed directly by chemical bonding or indirectly through a capturing substance. For indirect binding, for example, a combination of biotin and avidin, hapten and anti-hapten antibody, nickel and histatidine tag, glutathione and glutathione-S-transferase, etc. can be used. “Avidins” means containing avidin and streptavidin.

As an example, a fluorescent dye conjugated with biotin can be bound to a solid phase carrier coated with streptavidin. As such a solid phase carrier 22 to which the labeling substance 21 is bound, a commercially available one can be suitably used.

Further, by using a test substance having a known concentration instead of a specimen as a quality control sample (calibrator) and performing a measurement process using a measurement reagent for each measurement item, a solid phase to which the labeling substance 21 is bound. The carrier 22 may be formed. That is, the solid phase carrier 22 to which the labeling substance 21 is bound is prepared by forming an immune complex 84 composed of the labeling substance 83, the test substance 81, and the capture substance 86 on the solid phase carrier 82 shown in FIG. May be. In this case, as described later, such a substance can be produced by causing the immunoassay apparatus 100 to execute part or all of the immunoassay operation.

In FIG. 1, when the step (b) is performed, the labeling substance 21 in the second container 12 is detected. When the function of the complex transfer process of the immunoassay apparatus 100 is normal, the solid phase carrier 22 is not carried over to the second container 12 beyond the allowable upper limit for use. Therefore, when the function of collecting the solid phase carrier 22 of the immunoassay apparatus 100 is normal, the detection value of the labeling substance 21 in step (b) is as low as that obtained when a blank sample not containing the labeling substance 21 is measured. It becomes. Here, the blank sample is a sample that does not contain the labeling substance 21. On the other hand, when the function of collecting the solid phase carrier 22 of the immunoassay device 100 is abnormal, for example, when the solid phase carrier 22 is dispersed in the first container 11 without being collected, the step (a) The solid phase carrier 22 to which the labeling substance 21 is bound is transferred to the second container 12 together with the liquid phase. Therefore, when the function of collecting the solid phase carrier 22 of the immunoassay apparatus 100 is abnormal, the detected value of the labeling substance 21 in step (b) becomes an abnormally high value that is outside the allowable range of the detected value in the normal state.

For this reason, the detection result by the step (b) is acquired in the step (c). Based on the detection result, the state of the immunoassay apparatus 100 in the complex transfer process is determined. That is, it is determined from the detection result whether the complex transfer process has been successfully performed. The state of determination may be a binary classification of “normal” or “abnormal”, “0 (no abnormality)” or “1 (abnormal)”. As a state of determination, the degree of normality or abnormality may be determined according to the detection value. For example, within the normal state, set the judgment range such as within the complete normal range, outside the normal range but within the allowable range, within the allowable range but within the boundary range close to the abnormal value (outside the allowable range), and depending on the detection value It may be determined which range belongs. In this case, the user can obtain information for determining the necessity of maintenance of the immunoassay device 100, for example, based on the determination result.

(Control sample containing liquid in which labeling substance is dissolved)
As shown in FIG. 2, when the control sample 20 contains a liquid in which the labeling substance 21 is dissolved, the liquid phase containing the labeling substance 21 is transferred from the first container 11 to the second container 12 in step (a). . For this reason, the liquid phase containing the labeling substance 21 is accommodated in the second container 12.

In FIG. 2, when the step (b) is performed, the labeling substance 21 in the second container 12 is detected. When the function of transferring the liquid phase in the first container 11 of the immunoassay apparatus 100 to the second container 12 is normal, the labeling substance 21 in the liquid phase is sufficiently transferred into the second container 12. Therefore, when the function of transferring the liquid phase in the first container 11 of the immunoassay apparatus 100 to the second container 12 is normal, the detected value of the labeling substance 21 in step (b) is the labeling substance contained in the management sample 20. Within the tolerances expected from 21 known concentrations. On the other hand, when the function of transferring the liquid phase in the first container 11 of the immunoassay apparatus 100 to the second container 12 is abnormal, for example, the liquid phase could not be transferred from the first container 11 to the second container 12. In this case, the liquid phase containing the labeling substance 21 is not transferred to the second container 12 even by the step (a). Therefore, when the function of transferring the liquid phase in the first container 11 of the immunoassay apparatus 100 to the second container 12 is abnormal, the detection value of the labeling substance 21 in step (b) is determined from the known concentration of the labeling substance 21. The value deviates from the expected allowable range.

For this reason, in step (c), from the detection results obtained by performing steps (a) and (b) on the control sample 20 containing the liquid in which the labeling substance 21 is dissolved, immunoassay in complex transfer treatment is performed. The state of the device 100 is determined. That is, it is determined whether the complex transfer process has been successfully performed.

As described above, in the method for confirming the state of the immunoassay device according to the present embodiment, the same operation as the complex transfer process is performed on the first container 11 containing the management sample 20 including the labeling substance 21, and the second The labeling substance 21 in the control sample 20 transferred to the container 12 can be detected. If the complex transfer process is not performed normally, an abnormal detection result deviating from the expected detection result is acquired. Thereby, based on the detection result of the labeling substance 21 in the second container 12, it is possible to determine the state of the immunoassay device 100, such as whether the complex transfer process has been successfully performed. As a result, it is possible to easily check the status of the function for executing the immune complex transfer method in the immunoassay apparatus 100.

<Treatment of collecting the solid support>
As shown in FIG. 1, when the management sample 20 includes a solid phase carrier 22 to which a labeling substance 21 is bound, in the step (a) of transferring the liquid phase in the first container 11 to the second container 12, the first container 11. The solid phase carrier 22 to which the labeling substance 21 is bound is collected.

The process of collecting the solid phase carrier 22 is performed, for example, by centrifuging the first container 11 with the immunoassay apparatus 100 and allowing the solid phase carrier 22 to settle at the bottom of the container. When the solid support 22 is a magnetic particle, the process of collecting the solid support 22 includes a process of collecting the magnetic particles by the magnetic force source 52. For example, magnetic particles are collected by a magnetic source 52 provided in the immunoassay apparatus 100.

Thereby, since the solid phase carrier 22 to which the labeling substance 21 is bound is collected in the first container 11, for example, when the detection value of the labeling substance 21 increases, the solid phase is transferred when the liquid phase is transferred to the second container 12. There is a high possibility that the carrier 22 is not sufficiently collected and the solid phase carrier 22 is also transferred. Therefore, it can be easily confirmed whether or not the function of collecting the solid phase carrier 22 in the first container 11 in the complex transfer process is normal.

In addition to steps (a) and (b), there may be the following steps.

(Step of determining abnormality in complex transfer process)
For example, in FIGS. 1 and 2, the method for confirming the state of the immunoassay apparatus further includes a step of determining an abnormality in the complex transfer process based on the detection result of the labeling substance 21 in the second container 12. As described above, the presence or absence of abnormality in the complex transfer process can be determined by determining the magnitude of the detection value acquired in step (b). Thereby, for example, it is possible to determine whether or not there is an abnormality in the complex transfer process by determining the detection result in the immunoassay apparatus 100 that detects the labeling substance 21 in the management sample 20. The determination can be performed by, for example, a determination unit 70 (see FIG. 3) included in the immunoassay apparatus 100 or an external apparatus such as a host computer connected to the immunoassay apparatus 100 so as to be communicable.

Specifically, the function for performing the complex transfer process includes a function of collecting the solid phase carrier 22 in the first container 11 and a function of transferring the liquid phase in the first container 11 to the second container 12. Including. Therefore, the step of determining abnormality in the complex transfer process includes at least one of a function of collecting the solid phase carrier 22 in the first container 11 and a function of transferring the liquid phase in the first container 11 to the second container 12. Including determining any abnormality.

In the complex transfer process, the liquid phase containing the immune complex 84 is transferred to the second container 12, and the solid phase carrier 22 in the first container 11 is left in the first container 11 to leave the solid phase carrier 22. Whether the collecting function is normal or not is important. As shown in FIG. 1, when the management sample 20 includes the solid phase carrier 22 to which the labeling substance 21 is bound, the solid phase carrier 22 in the first container 11 is changed based on the detection result of the step (b). It is possible to determine whether or not the function to be collected is normal.

In the complex transfer process, whether or not the function of transferring the liquid phase in the first container 11 to the second container 12 is normal in order to detect the immune complex 84 transferred to the second container 12 is determined. It becomes important. As shown in FIG. 2, when the management sample 20 includes a liquid in which the labeling substance 21 is dissolved, the liquid phase in the first container 11 is transferred to the second container 12 based on the detection result of the step (b). It can be determined whether or not the function to be transferred is normal. According to the above configuration, since it is possible to determine the presence / absence of at least one of these functions, it is particularly useful for ensuring the reliability of the measurement result. In order to ensure high reliability, both the determination of the function of collecting the solid phase carrier 22 in the first container 11 and the determination of the function of transferring the liquid phase in the first container 11 to the second container 12 are both performed. Are preferably performed.

When performing the step of determining an abnormality in the complex transfer process, the abnormality is detected based on whether the detected value of the labeling substance 21 in the second container 12 exceeds the reference value V1 or the detected value is lower than the reference value V1. judge. Thus, by setting a reference value V1 for distinguishing between normal and abnormal from the detection value obtained by measuring the control sample 20 in advance, the composite value can be obtained simply by comparing the detection value with the reference value V1. Abnormality determination of the transfer process can be easily performed.

<Process for dispensing the control sample into the first container>
In FIG. 1 and FIG. 2, before the step of transferring the liquid phase in the first container 11 to the second container 12, a step of dispensing the management sample 20 into the first container 11 by the immunoassay device 100 may be further provided. . For example, the immunoassay apparatus 100 sucks the management sample 20 from a container (not shown) that stores the management sample 20 in advance by the suction tube 51 and discharges the management sample 20 into the first container 11. Thereby, even if the user or service staff of the immunoassay apparatus 100 does not prepare the management sample 20 in the first container 11 in advance, the management sample 20 is easily accommodated in the first container 11 by the immunoassay apparatus 100. Can be made.

[Outline of immunoassay device]
Next, with reference to FIG. 3, the outline | summary of the immunoassay apparatus 100 by one Embodiment is demonstrated.

The immunoassay device 100 is a device that measures a test substance in a specimen using an antigen-antibody reaction. The immunoassay apparatus 100 performs a complex transfer process by an immune complex transfer method. That is, as shown in FIG. 4, the immunoassay device 100 includes an inside of the first container 11 that contains an immune complex 84 that includes a test substance 81 and a labeling substance 83 in a sample and is carried on a solid phase carrier 82. Thus, the immunoassay apparatus 100 performs the complex transfer process in which the immune complex 84 is released from the solid phase carrier 82 and the liquid phase in the first container 11 is transferred to the second container 12.

As shown in FIG. 3, the immunoassay apparatus 100 includes a mechanism unit 50 that performs a process of transferring the liquid phase in the first container 11 in which the management sample 20 containing at least the labeling substance 21 is stored to the second container 12, and the liquid The detection part 60 which detects the labeled | labeling substance 21 in the 2nd container 12 in which the phase was transferred, and the determination part 70 which determines the abnormality in a complex transfer process based on the detection result of the detection part 60 are provided.

The mechanism unit 50 has at least a function of executing a complex transfer process. That is, the mechanism unit 50 is configured to perform a process of transferring the liquid phase in the first container 11 to the second container 12. The mechanism unit 50 may have a function of performing not only the complex transfer process but also a process necessary for the immunoassay on the reaction container. For example, the mechanism unit 50 can perform a process of forming an immune complex including the test substance 81 and the labeling substance 21 in the sample on the solid phase carrier 22 in the first container 11.

The mechanism unit 50 can include one or a plurality of processing units depending on the type and number of processing steps performed by the immunoassay apparatus 100. One processing unit may perform one type of processing step, or may be a processing unit capable of performing a plurality of types of processing steps.

The mechanism unit 50 includes, for example, a suction pipe 51 that sucks the liquid phase in the first container 11 and discharges the sucked liquid phase to the second container 12. The suction pipe 51 is connected to a pressure source (not shown) such as a metering pump, for example, and sucks a predetermined amount of liquid from the tip and can dispense a fixed amount.

When the management sample 20 includes the solid phase carrier 22 to which the labeling substance 21 is bound, the mechanism unit 50 includes a magnetic force source 52 that collects magnetic particles bound to the labeling substance 21 in the first container 11. The magnetic source 52 is positioned in the vicinity of the first container 11. Magnetic particles can be collected by the magnetic force of the magnetic source 52. The magnetic collection is to collect magnetic materials by applying a magnetic force. For example, the magnetic force source 52 applies a magnetic force to the magnetic particles in the first container 11 to collect the magnetic particles at a predetermined position such as the inner surface or the bottom of the first container 11. As the magnetic source 52, for example, a permanent magnet or an electromagnet can be employed.

For example, the mechanism unit 50 may include a sample dispensing unit for dispensing the sample into the first container 11. In this case, the mechanism unit 50 can perform a step of dispensing a specimen containing the test substance 81. When the mechanism unit 50 processes the first container 11 in which the sample has been dispensed in advance, it is not necessary to provide the sample dispensing unit in the mechanism unit 50.

Further, the mechanism unit 50 may include a reagent dispensing unit for dispensing the reagent to the first container 11. In this case, the mechanism unit 50 includes a step of dispensing a solid phase reagent including

solid phase carriers

22 and 82 such as magnetic particles, a step of dispensing a labeling reagent including

labeling substances

21 and 83, and a free reagent 85. A dispensing process can be performed. When the mechanism unit 50 processes the first container 11 in which these reagents are dispensed in advance, it is not necessary to provide the reagent dispensing unit in the mechanism unit 50.

The various reagents dispensed into the first container 11 are liquid reagents and are stored in separate reagent containers for each type. The solid phase reagent is a liquid reagent containing

solid phase carriers

22 and 82 such as magnetic particles in a liquid, and the labeling reagent is a liquid reagent containing

labeling substances

21 and 83 in the liquid. The free reagent 85 is a liquid reagent containing a component for eliminating the binding between the immune complex 84 and the solid phase carrier 82.

The release reagent 85 eliminates the binding between the immune complex 84 including the test substance 81 and the labeling substance 83 and the solid phase carrier 82, and releases the immune complex 84 from the solid phase carrier 82. When the solid phase carrier 82 and the test substance 81 are bound to each other, the free reagent 85 cancels the binding between the solid phase carrier 82 and the test substance 81. When the solid phase carrier 82 binds to the test substance 81 via the capture substance 86, the free reagent 85 binds to the solid phase carrier 82 and the capture substance 86, or the test substance 81 and the capture substance 86 bind to each other. Can be eliminated. The free reagent 85 is selected according to the type of binding between the immune complex 84 and the solid phase carrier 82.

For example, when the binding between the immune complex 84 and the solid phase carrier 82 is binding by a hapten-anti-hapten antibody, a hapten or a hapten derivative can be used as the free reagent 85. Further, when the bond between the immune complex 84 and the solid phase carrier 82 is a bond by ionic bond, a solution containing ions can be used as the free reagent 85. Further, when the binding between the immune complex 84 and the solid phase carrier 82 is a binding by a ligand-receptor as a separable binding, a ligand or a ligand analog can be used as the free reagent 85. When the bond between the immune complex 84 and the solid phase carrier 82 is a lectin-sugar chain bond as a separable bond, a carbohydrate can be used as the free reagent 85. When the immune complex 84 and the solid phase carrier 82 are bound by biotin-avidin, biotin can be used as the free reagent 85.

Further, the mechanism unit 50 may include a reaction unit for heating and reacting the sample in the first container 11. In this case, the mechanism unit 50 can promote the reaction of the sample in a temperature environment suitable for the reaction during the process of forming the immune complex 84 or the process of releasing the immune complex 84, so that the process is efficiently performed. Can do. When the reaction proceeds sufficiently without heating the sample in the first container 11, or when the entire mechanism unit 50 is configured as a constant temperature bath at a predetermined temperature, the reaction unit is provided in the mechanism unit 50. There is no need.

The detection unit 60 has a function of detecting the

labeling substances

21 and 83 in the liquid phase dispensed into the second container 12. As described above, the detection may be performed by an appropriate method according to the types of the

labeling substances

21 and 83, and the detection method is not particularly limited. As the detection unit 60, a photomultiplier tube, a spectrophotometer, a luminometer, or the like can be used. When detecting fluorescence, the detection unit 60 may include a light source for irradiating excitation light. Further, when the labeling substance is a radioisotope, a scintillation counter or the like can be used as the detection unit 60.

The determination unit 70 acquires the detection result of the detection unit 60. The determination unit 70 is configured by a computer including a processor such as a CPU and a storage unit such as a hard disk drive and a flash memory, for example. The processor functions as the determination unit 70 of the immunoassay device 100 by executing the control program stored in the storage unit. The determination unit 70 determines an abnormality in the complex transfer process based on the detection result of the detection unit 60.

The immunoassay device 100 according to the present embodiment performs the state confirmation method of the immunoassay device 100 shown in at least one of FIGS. 1 and 2 with the above-described configuration. Thereby, the liquid phase transfer operation similar to the complex transfer process is performed on the first container 11 containing the management sample 20 including the labeling substance 21, and the management sample 20 transferred to the second container 12. The labeling substance 21 inside can be detected. If the complex transfer process is not performed normally, an abnormal detection result deviating from the expected detection result is acquired. Thereby, it can be confirmed from the detection result of the labeling substance 21 in the second container 12 whether or not the complex transfer process has been normally performed, so that the immune complex transfer method in the immunoassay device is executed. The function status can be easily checked.

[Specific configuration example of immunoassay device]
Next, a specific configuration example of the immunoassay device 100 will be described in detail with reference to FIG.

The immunoassay apparatus 100 includes a mechanism unit 50, a detection unit 60, and a determination unit 70. In the configuration example of FIG. 5, the immunoassay device 100 includes an analysis unit 110 for analyzing an immunoassay result. The determination unit 70 is realized as part of the function executed by the analysis unit 110.

The mechanism unit 50 includes a sample dispensing unit 120, a reagent dispensing unit 130, a container supply unit 140, a reagent storage 150, a reaction unit 160, and a BF separation unit 170. Moreover, the mechanism part 50 contains the container transfer part 180 which conveys a container to each of these parts. The immunoassay apparatus 100 includes a sample transport unit 190 and a housing 105 that houses the mechanism unit 50 and the detection unit 60.

The housing 105 has a box shape that houses each part of the immunoassay device 100 therein. The housing 105 can be configured with one or a plurality of layers.

The sample transport unit 190 is configured to transport a sample collected from the subject to a suction position by the sample dispensing unit 120. The sample transport unit 190 can transport a rack in which a plurality of sample containers 191 (see FIG. 6) containing a sample are installed to a predetermined sample suction position.

The sample dispensing unit 120 can aspirate the sample conveyed by the sample conveyance unit 190 and dispense the aspirated sample into the first container 11. As shown in FIG. 6, the sample dispensing unit 120 includes a suction tube 121 connected to a fluid circuit for performing suction and discharge, and a moving mechanism (not shown) that moves the suction tube 121. The sample dispensing unit 120 attaches a dispensing tip 122 to the tip of the suction tube 121 from a tip supply unit (not shown), for example, and sucks a predetermined amount of the sample in the transported sample container 191 into the dispensing tip 122. The sample dispensing unit 120 dispenses the aspirated sample into the first container 11 disposed at a predetermined sample dispensing position. After dispensing, the sample dispensing unit 120 removes the dispensing tip 122 from the tip of the suction tube 121 and discards it.

The container supply unit 140 can store a plurality of unused reaction containers. That is, the container supply unit 140 can store a plurality of unused first containers 11 and second containers 12 and supply them to predetermined container supply positions. In this configuration example, reaction containers having the same shape and the same material are used as the first container 11 and the second container 12. That is, the unused reaction container supplied from the container supply unit 140 can be used as both the first container 11 and the second container 12. In addition, when the 1st container 11 and the 2nd container 12 correspond in common, and it is not necessary to distinguish, it is only called "reaction container 10."

The container transfer unit 180 can transfer the reaction container 10. The container transfer unit 180 acquires an empty container from the container supply position, and puts the container at each processing position such as the sample dispensing unit 120, the reagent dispensing unit 130, the reaction unit 160, the BF separation unit 170, the detection unit 60, and the like. Transport. The container transfer unit 180 includes, for example, a catcher that holds the container or a holding unit having a container installation hole, and a moving mechanism that moves the catcher or the holding unit. The moving mechanism moves in the direction of one axis or a plurality of axes by, for example, one or more linear motion mechanisms capable of linear movement. The moving mechanism can move, for example, in three orthogonal directions in the vertical direction and two horizontal directions. The moving mechanism may include an arm mechanism that rotates horizontally around the rotation axis and an articulated robot mechanism. One or a plurality of container transfer units 180 are provided according to the arrangement of the processing positions of the units in the housing 105.

The reaction unit 160 includes a heater and a temperature sensor, holds the reaction vessel 10, and heats and reacts the sample stored in the vessel. By heating, the specimen and reagent contained in the container react. One or more reaction units 160 are provided in the housing 105. The reaction unit 160 may be fixedly installed in the housing 105 or may be movably provided in the housing 105. When the reaction unit 160 is configured to be movable, the reaction unit 160 can also function as a part of the container transfer unit 180.

The reagent storage 150 has a box shape and includes a container holding portion 151 and a cooling mechanism inside. The container holding unit 151 holds the reagent container 155. The cooling mechanism keeps the reagent in the reagent container 155 at a constant temperature suitable for storage. The reagent storage 150 has a plurality of holes 152 on the upper surface for allowing the reagent dispensing unit 130 to enter the inside of the reagent storage 150.

The container holding part 151 is formed to hold a plurality of reagent containers 155 side by side in the circumferential direction. The container holding part 151 can hold a plurality of reagent containers 155 side by side in the radial direction. That is, the container holding part 151 can arrange the row | line | column of the some reagent container 155 arranged in a circle by arranging in the radial direction concentrically. The container holding part 151 can independently rotate the row of a plurality of concentric reagent containers 155 in the circumferential direction. As a result, the container holding unit 151 has a desired reagent container selected from the row of the corresponding reagent containers 155 at a position immediately below each of the plurality of holes 152 provided in accordance with the reagent dispensing unit 130. 155 can be arranged. As a result, the reagent in the reagent container 155 arranged at a position immediately below the hole 152 is aspirated by the reagent dispensing unit 130. In the container holding part 151, a reagent container 155 that stores r1 reagent, r4 reagent, r5 reagent, r6 reagent, and r7 reagent, which will be described later, is set. The reagent container 155 may have a structure that has a plurality of storage chambers and can store a plurality of types of reagents.

The reagent dispensing unit 130 sucks the reagent in the reagent container 155 and dispenses the sucked reagent into the reaction container 10. The reagent dispensing unit 130 can move a suction tube 131 for performing aspiration and discharge of the reagent in the horizontal direction between the hole 152 and a predetermined reagent dispensing position. In addition, as shown in FIG. 7, the reagent dispensing unit 130 can move the suction tube 131 in the vertical direction and pass through the hole 152 from above the hole 152 to enter the reagent container 155. The suction tube 131 can be retracted to a position above the hole 152. The suction tube 131 is connected to a fluid circuit (not shown), sucks a predetermined amount of reagent from the reagent container 155 of the container holding unit 151, and dispenses the reagent into the reaction container 10 transferred to the reagent dispensing position.

For example, one or a plurality of reagent dispensing units 130 are provided. In the example of FIG. 5, three reagent dispensing units 130 are provided on the reagent storage 150. Each of the three reagent dispensing units 130 is set in advance as to which of the r1 reagent, r4 reagent, r5 reagent, r6 reagent, and r7 reagent is to be dispensed. There is no particular limitation on whether or not to add. The mechanism unit 50 includes an R4 reagent dispensing unit 134 for dispensing the R4 reagent and an R5 reagent dispensing unit 135 for dispensing the R5 reagent.

The R4 reagent dispensing unit 134 and the R5 reagent dispensing unit 135 are provided at positions separated from the reagent storage 150. The R4 reagent dispensing unit 134 and the R5 reagent dispensing unit 135 are connected to a reagent container (not shown) containing the R4 reagent and the R5 reagent via a liquid feeding tube, respectively, and are transferred by the container transfer unit 180. The reagent can be discharged into the reaction container 10.

The BF separation unit 170 has a function of executing BF separation for separating the liquid phase and the solid phase from the reaction vessel 10. In the immunoassay apparatus 100, one or a plurality of BF separation units 170 are provided. As shown in FIG. 8, the BF separation unit 170 sucks the liquid component in the reaction vessel 10 with the suction tube 171 while collecting the magnetic particles with the magnetic collection unit 173, and supplies the cleaning liquid with the discharge tube 172 To do. The suction pipe 171 and the discharge pipe 172 are each connected to a fluid circuit (not shown). Thereby, the unnecessary substance contained in the liquid component can be separated and removed from the magnetic particles.

The mechanism unit 50 has a function of performing a complex transfer process.

As shown in FIGS. 9A to 9C, the mechanism unit 50 includes a magnetic source 52 that collects magnetic particles to which the labeling substance 21 is bound in the first container 11. The magnetic force source 52 is provided in the holding member 210 in which a holding hole 211 for holding the first container 11 is formed, for example. The magnetic source 52 is constituted by a permanent magnet, for example. The holding member 210 may be fixedly installed in the housing 105, may be configured to be movable, and may function as a part of the container transfer unit 180.

The mechanism unit 50 includes a suction pipe 51 that sucks the liquid phase in the first container 11 and discharges the sucked liquid phase to the second container 12. The suction tube 51 sucks the liquid phase from the first container 11 held by the holding member 210 and puts it into the second container 12 held by the container transfer part 180 or another container holding part (not shown). Dispense the sucked liquid phase.

The liquid phase transfer in the complex transfer process is performed by a configuration including a suction tube. For example, as shown in FIG. 9A, the complex transfer process can be executed by the suction tube 121 of the sample dispensing unit 120. The suction tube 121 also functions as the suction tube 51 that performs the complex transfer process. As shown in FIG. 9B, the complex transfer process can be executed by the suction tube 131 of the reagent dispensing unit 130. The suction tube 131 also functions as the suction tube 51 that performs the complex transfer process. As shown in FIG. 9C, the mechanism unit 50 may include a dedicated immune complex dispensing unit 220 for performing the complex transfer process. In that case, the liquid phase in the first container 11 is sucked by the suction tube 51 provided in the immune complex dispensing unit 220, and the sucked liquid phase is discharged to the second container 12.

As shown in FIG. 10, the detection unit 60 includes a photodetector 61 such as a photomultiplier tube. The detection unit 60 receives the second container 12 inside, and detects light generated in the reaction process of the labeling substance 83 (see FIG. 4) that binds to the analyte 81 of the specimen and the luminescent substrate by the photodetector 61. . The detection unit 60 counts the number of photons detected by the photodetector 61 and outputs it as a detection value.

Returning to FIG. 5, the analysis unit 110 is configured by a personal computer, for example. The analysis unit 110 includes, for example, a processor 111 such as a CPU and a storage unit 112 such as a ROM, a RAM, and a hard disk. The processor 111 functions as the analysis unit 110 of the immunoassay device 100 by executing the control program stored in the storage unit 112.

The analysis unit 110 is electrically connected to the mechanism unit 50, and controls the mechanism unit 50 so as to execute measurement of a sample and confirmation of the state of the immunoassay device 100. The analysis unit 110 causes the mechanism unit 50 to perform sample measurement according to the measurement order acquired from a host computer (not shown), and analyzes the detection result by the detection unit 60. That is, the analysis unit 110 compares the detection value obtained by the detection unit 60 with a calibration curve prepared in advance, so that the abundance of the test substance 81 in the sample (that is, the labeling substance bound to the test substance 81). 83 abundance). The analysis unit 110 includes a determination unit 70. That is, the analysis unit 110 also functions as the determination unit 70 when the processor 111 executes the control program. A dedicated processor that functions as the determination unit 70 may be provided.

The determination unit 70 is at least one of the function of collecting the solid phase carrier 22 in the first container 11 by the mechanism unit 50 and the function of transferring the liquid phase in the first container 11 by the mechanism unit 50 to the second container 12. Judge abnormalities. Accordingly, since it is possible to determine whether or not there is an abnormality in at least one of the function of collecting the solid phase carrier 22 in the complex transfer process and the function of transferring the liquid phase to the second container 12, the reliability of the measurement result is ensured. Is particularly useful.

Specifically, the determination unit 70 determines an abnormality in the magnetic collection function by the magnetic force source 52 (see FIGS. 9A to 9C) based on the detection result for the management sample including the magnetic particles to which the labeling substance 21 is bound. As a result, the solid phase carrier 22 to which the labeling substance 21 is bound is collected in the first container 11. For example, when the detection value of the labeling substance 21 increases, the liquid phase is transferred to the second container 12. There is a high possibility that the solid phase carrier 22 is not sufficiently magnetized and the solid phase carrier 22 is also transferred. Therefore, it can be easily determined whether the magnetic flux collecting function by the magnetic source 52 in the composite transition process is normal or abnormal.

In addition, the determination unit 70 determines abnormality of the dispensing function by the suction tube 51 (see FIGS. 9A to 9C) based on the detection result for the management sample including the solution in which the labeling substance 21 is dissolved. Thereby, since the labeling substance 21 exists in the first container 11 as the liquid phase, for example, when the detection value of the labeling substance 21 does not increase, the liquid phase is dispensed into the second container 12. There is a high possibility that the dispenser by the suction pipe 51 that performs the suction and the liquid phase discharge is not normal. Therefore, the presence or absence of abnormality in the dispensing function by the suction tube 51 in the complex transfer process can be easily confirmed.

(Overview of immunoassay)
In the configuration example shown in FIG. 5, as shown in FIG. 4, immunoassay is performed using the r1 reagent to r7 reagent, the R4 reagent, and the R5 reagent. Here, an example in which the test substance 81 is hepatitis B surface antigen (HBsAg) will be described as an example of immunoassay.

First, the reagent dispensing unit 130 dispenses the r1 reagent into the first container 11. The r1 reagent is a labeling reagent containing the labeling substance 83. The labeling substance 83 reacts with and binds to the test substance 81. In the example of FIG. 4, the labeling substance is an ALP (alkaline phosphatase) labeled antibody. After dispensing of each reagent, the sample in the reaction container 10 is heated to a predetermined temperature in the reaction unit 160 for a predetermined time. In the following description, description of the reaction unit 160 is omitted.

Next, the specimen dispensing unit 120 dispenses a specimen containing the test substance 81 in the first container 11. The test substance 81 binds to the labeling substance 83. Depending on the test substance 81, a reagent (r2 reagent) for alkali denaturation of the specimen as a pretreatment for releasing the antigen present in the specimen in a state in which the antibody is bound in advance from the antibody, or the alkali of the specimen is neutralized. A reagent (r3 reagent) or the like may be further dispensed.

Next, the reagent dispensing unit 130 dispenses the r4 reagent into the first container 11. The r4 reagent is a capture reagent containing a capture substance 86 that reacts with and binds to the test substance 81. The capture substance 86 includes a first binding substance 86a for binding the capture substance 86 to a first solid phase carrier 82a described later, and a second binding substance for binding the capture substance 86 to a second solid phase carrier 82b described later. 86b. The first binding substance 86a and the second binding substance 86b are substances that bind to the solid phase carrier with different binding abilities.

In the example of FIG. 4, the capture substance 86 is an antibody (DNP / biotin antibody) modified with DNP (dinitrophenyl group) and biotin. That is, the capture substance 86 is modified with DNP (dinitrophenyl group) as the first binding substance 86a and biotin as the second binding substance 86b.

Next, the reagent dispensing unit 130 dispenses the r5 reagent into the first container 11. The r5 reagent is a solid phase reagent containing a solid phase carrier 82. Specifically, the r5 reagent contains the first solid phase carrier 82a as the solid phase carrier 82. The first solid phase carrier 82a is a magnetic particle, specifically, a magnetic particle to which an anti-DNP antibody is immobilized (anti-DNP antibody-modified magnetic particle). The anti-DNP antibody-antimagnetic DNP antibody, which is an anti-hapten, reacts with and binds to the DNP of the capture substance 86, which is a hapten. As a result, an immune complex 84 including the test substance 81, the labeling substance 83, and the capture substance 86 is formed on the first solid phase carrier 82a.

The immune complex 84 formed on the first solid phase carrier 82a and the unreacted labeling substance 83 are separated by the primary BF separation process. Unnecessary components such as the unreacted labeling substance 83 are removed from the first container 11 by the primary BF separation process. The primary BF separation process is performed by the BF separation unit 170 (see FIG. 8).

After the primary BF separation process, the reagent dispensing unit 130 dispenses the r6 reagent into the first container 11. The r6 reagent is the free reagent 85. In the example of FIG. 4, DNP-Lys (DNP-Lysine) is used as the free reagent 85. DNP-Lys reacts and binds to the anti-DNP-antibody magnetic particles that are the first solid phase carrier 82a. Therefore, when the r6 reagent is dispensed into the first container 11, the binding between the DNP of the capture substance 86 and the first solid phase carrier 82a, the free reagent 85 (DNP-Lys), and the first solid phase carrier 82a. The binding competes and the binding between the capture substance 86 and the first solid phase carrier 82a is eliminated. As a result, the immune complex 84 is released from the first solid phase carrier 82a.

Next, a complex transfer process is performed. That is, the liquid phase containing the immune complex 84 released by the r6 reagent is sucked from the first container 11 by the suction tube 51 and dispensed into the second container 12.

By the complex transfer treatment, the liquid phase containing the immune complex 84 released from the first solid phase carrier 82a is transferred from the first container 11 to the second container 12. The first solid phase carrier 82a remains in the first container 11 after the liquid phase containing the immune complex 84 is aspirated. As a result, the labeling substance 83 nonspecifically bound to the first solid phase carrier 82a is separated from the immune complex 84.

The reagent 7 is then dispensed with the r7 reagent into the second container 12 into which the immune complex 84 has been dispensed. The r7 reagent contains the second solid phase carrier 82b. The second solid phase carrier 82b binds to the second binding substance 86b of the capture substance 86. The second solid phase carrier 82b is a magnetic particle, specifically, a magnetic particle (StAvi-binding magnetic particle) to which streptavidin that binds to biotin is immobilized. StAvi-bound magnetic particles streptavidin reacts with and binds to biotin as the second binding substance 86b. As a result, the immune complex 84 including the test substance 81, the labeling substance 83, and the capture substance 86 is bound to the second solid phase carrier 82b.

The immune complex 84 bound to the second solid phase carrier 82b and unnecessary components other than the second solid phase carrier 82b on which the immune complex 84 is formed are separated by the secondary BF separation process, and the unnecessary components are secondly separated. The container 12 is removed. The unnecessary component is, for example, a free reagent 85 contained in the liquid phase, a labeling substance 83 contained in the liquid phase together with the immune complex 84 without being bound to the test substance 81, and the like. The secondary BF separation process is performed by the BF separation unit 170 (see FIG. 8).

Thereafter, the R4 reagent dispensing unit 134 and the R5 reagent dispensing unit 135 respectively dispense the R4 reagent and the R5 reagent into the second container 12. The R4 reagent contains a buffer. The immune complex 84 bound to the second solid phase carrier 82b is dispersed in the buffer. The R5 reagent contains a chemiluminescent substrate. The buffer solution contained in the R4 reagent has a composition that promotes the reaction between the label (enzyme) of the labeling substance 83 contained in the immune complex 84 and the substrate. Light is generated by reacting the substrate with the label, and the intensity of the generated light is measured by the detection unit 60 (see FIG. 10).

(Immunoassay device status confirmation method)
Next, a state confirmation method in a specific configuration example of the immunoassay apparatus 100 illustrated in FIG. 5 will be described.

In the following example, the labeling substance 21 contained in the control sample 20 is an enzyme. The step (b) of detecting the labeling substance 21 in the second container 12 is performed by adding an enzyme substrate to the second container 12 and measuring a signal generated from a reaction product generated by the enzyme reaction.

<Confirmation of magnetism collecting function>
With reference to FIG. 12 to FIG. 14, an example in which the magnetism collecting function of the immunoassay apparatus 100 is confirmed will be shown. 12 to 14, the management sample 20 includes a solid phase carrier 22 to which a labeling substance 21 is bound, and the solid phase carrier 22 is a magnetic particle. The process of collecting the solid phase carrier 22 includes a process of collecting magnetic particles by the magnetic force source 52. The state confirmation method includes a step of determining abnormality of the magnetic flux collecting function by the magnetic source 52 based on the detection result of the labeling substance 21 in the second container 12. Thereby, in order to collect the magnetic particles to which the labeling substance 21 is bound, whether the magnetism collection function by the magnetic source 52 is normal or abnormal is determined based on the detection value of the labeling substance 21 in the second container 12. Easy to judge.

Specifically, as shown in FIG. 11, when the detection value of the labeling substance 21 exceeds the reference value V1, the determination unit 70 determines that there is an abnormality in the magnetic collection function by the magnetic source 52. That is, when the detection value of the labeling substance 21 exceeds the reference value V1, there is a high possibility that the magnetic particles cannot be sufficiently collected when the liquid phase is transferred to the second container 12. Thereby, the magnetism collecting function by the magnetic source 52 can be easily determined by comparing the detected value with the reference value V1.

The reference value V1 when confirming the magnetism collecting function is an expected detection value of the labeling substance 21 corresponding to the allowable upper limit amount of the magnetic particles that are transferred to the second container 12 without being magnetized. Thereby, when the amount of magnetic particles exceeding the allowable upper limit amount is transferred to the second container 12, it can be easily determined that the magnetic flux collecting function by the magnetic force source 52 is abnormal. Therefore, the reliability of the measurement result can be ensured by confirming that the allowable upper limit amount is not exceeded.

(Embodiment 1)
In the example of FIG. 12, the management sample 20 includes a third solid phase carrier 23 that does not bind to the test substance 81 but binds to the labeling substance 21. In the liquid phase, the labeling substance 21 is not substantially contained. The third solid phase carrier 23 is, for example, a magnetic particle to which the labeling substance 21 is fixed in advance. The third solid phase carrier 23 does not have a binding ability to the test substance 81. That is, the third solid phase carrier 23 when confirming the magnetism collecting function is the first solid phase carrier 82a included in the r5 reagent when the sample is measured, and the second solid phase carrier included in the r7 reagent. It is included in the quality control reagent prepared separately from 82b.

Thus, the function of collecting the solid phase carrier 22 can be confirmed using the third solid phase carrier 23 not containing the test substance 81 as a dedicated reagent for checking the state of the immunoassay apparatus 100. When the solid phase carrier 22 is bound not only to the labeling substance 21 but also to the test substance 81, a plurality of types of solid phase carriers 22 and labeling substances 21 that can be bound according to the type of the test substance 81 are prepared. With the third solid phase carrier 23 not including the test substance 81, the state of the immunoassay apparatus 100 can be confirmed regardless of the type of the test substance 81. Therefore, the state of the immunoassay device 100 can be confirmed more easily.

In the example of FIG. 12, the management sample 20 including the third solid phase carrier 23 bound to the labeling substance 21 is accommodated in the first container 11. The management sample 20 is dispensed into the first container 11 by the mechanism unit 50. For example, the control sample 20 is set in the sample transport unit 190 while being stored in a predetermined control sample container in the same manner as the sample container 191, and is dispensed into the first container 11 by the sample dispensing unit 120. Further, for example, the management sample 20 is set in the reagent storage 150 while being accommodated in a predetermined management sample container in the same manner as the reagent container 155, and is dispensed into the first container 11 by the reagent dispensing unit 130.

During the step (a) of transferring the liquid phase in the first container 11 containing the management sample 20 to the second container 12, the third solid phase carrier 23 in the first container 11 is Magnetized in the container 11. The liquid phase in the first container 11 is sucked and dispensed into the second container 12 by the suction pipe 51 in the state of being magnetized. The third solid phase carrier 23 remains collected in the first container 11.

Next, a step (b) of detecting the labeling substance 21 in the second container 12 to which the liquid phase has been transferred is performed by the detection unit 60. By the step (b), a detection value indicating the abundance of the labeling substance 21 is acquired.

In the example of FIG. 12, the dispensing operation of the r1 reagent, the specimen, the r4 reagent, the r5 reagent, the r6 reagent, and the r7 reagent shown in FIG. 4 can be skipped. When the dispensing operation of the immunoassay apparatus 100 is not skipped, an alternative liquid that does not cause an antigen-antibody reaction, such as a buffer solution, may be dispensed instead of each reagent and sample. In the example of FIG. 12, the primary BF separation process and the secondary BF separation process shown in FIG. 4 are skipped. When dispensing the alternative liquid, BF separation may also be performed. FIG. 12 shows an example in which the management sample 20 is dispensed at the dispensing timing of the r6 reagent shown in FIG. 4, but the management sample 20 is dispensed at any timing before the complex transfer process. May be.

(Embodiment 2)
In the example of FIG. 13, the state confirmation method includes a sample containing a test substance 81 having a known concentration in the first container 11, a sample containing the test substance 81 before the step of transferring the liquid phase in the first container 11 to the second container 12. A step of contacting a labeling substance 83 that binds to the test substance 81, a capture substance 86 that binds to the test substance 81, and a solid phase carrier 82 that binds to the capture substance 86; That is, also in the state confirmation method, as in the case of measuring the specimen, the quality control sample including the test substance 81 and various reagents are dispensed into the first container 11 to prepare the control sample 20. As a result, the management sample 20 includes the solid phase carrier 82 bound to the immune complex 84 including the test substance 81, the labeling substance 83, and the capture substance 86. That is, FIG. 13 shows an example in which the labeling substance 83 and the solid phase carrier 82 used in actual sample measurement are used as the labeling substance 21 and the solid phase carrier 22 used in the state confirmation method of the immunoassay apparatus 100.

Thereby, the solid phase carrier 82 to which the labeling substance 83 is bound can be formed in the first container 11 by the same processing steps as in the case of actually measuring the specimen including the test substance 81. In the liquid phase, the labeling substance 83 is substantially not contained.

In the example of FIG. 13, first, the r1 reagent containing the labeling substance 83 is dispensed into the first container 11. Next, a quality control sample containing a test substance 81 having a known concentration is dispensed. Next, the r4 reagent containing the capture substance 86 is dispensed. Then, the r5 reagent containing the first solid phase carrier 82a is dispensed. As a result, the immune complex 84 including the test substance 81, the labeling substance 83, and the capture substance 86 is bound to the first solid phase carrier 82a. Reagent dispensing is performed by the reagent dispensing unit 130, and dispensing of the quality control sample is performed by the sample dispensing unit 120.

Thus, in the example of FIG. 13, the management sample 20 includes the first solid phase carrier 82a dispensed into the first container 11 before the complex transfer process at the time of measurement of the specimen. That is, the control sample 20 includes the first solid phase carrier 82a that is a magnetic particle contained in the r5 reagent. Thereby, the solid phase carrier 82 to which the labeling substance 83 is bound is formed in the first container 11 by using the first solid phase carrier 82a dispensed when the specimen including the test substance 81 is actually measured. be able to. That is, since it is not necessary to separately prepare a dedicated solid phase carrier 22 for confirming the state of the immunoassay device 100, the convenience of the immunoassay device 100 can be improved.

Next, primary BF separation processing is performed by the BF separation unit 170. That is, in the example of FIG. 13, the state confirmation method includes an immune complex including the test substance 81, the labeling substance 83, and the capture substance 86 before the step of transferring the liquid phase in the first container 11 to the second container 12. And a step of performing BF separation for separating the solid phase carrier 82 bonded to 84 and the liquid phase. Thereby, the labeling substance 83 in the liquid phase that has not been bonded to the solid phase carrier 82 can be separated and removed from the solid phase carrier by BF separation. Therefore, in the step of transferring the liquid phase in the first container 11 to the second container 12, it is possible to prevent the labeling substance 83 from being mixed in the liquid phase, so that the detection of the labeling substance 83 for confirming the state of the immunoassay device 100 is detected. Accuracy can be improved.

After the primary BF separation process, the reagent phase dispensing unit 130 dispenses the liquid phase 25 that does not contain the free reagent 85 into the first container 11. That is, when the sample is measured, the immune complex 84 is released from the solid phase carrier 22 by dispensing the free reagent 85 into the first container 11 in the complex transfer process. On the other hand, in the example of FIG. 13, the state confirmation method does not include the free reagent 85 after the step of performing BF separation and before the step of transferring the liquid phase in the first container 11 to the second container 12. The liquid phase is dispensed into the first container 11.

The liquid phase 25 that does not contain the free reagent 85 is not particularly limited, but is, for example, a buffer solution. As a result, the state of the immunoassay device 100 is confirmed by the same operation as dispensing the free reagent 85 during actual sample measurement. In this case, since the liquid phase 25 does not contain the free reagent 85, the labeling substance 83 and the solid phase carrier 82 remain bonded.

Next, a complex transfer process is performed. During the step (a) of transferring the liquid phase in the first container 11 containing the management sample 20 to the second container 12, the first solid phase carrier 82 a in the first container 11 is Magnetized in the container 11. The liquid phase in the first container 11 is sucked and dispensed into the second container 12 by the suction pipe 51 in the state of being magnetized. The first solid phase carrier 82a is left magnetized in the first container 11. Since the immune complex 84 remains bound to the first solid phase carrier 82a, the liquid phase not containing the labeling substance 83 is transferred to the second container 12.

Next, the reagent dispensing unit 130 dispenses the r7 reagent including the second solid phase carrier 82b into the second container 12 to which the liquid phase has been transferred. Then, the BF separation unit 170 performs a secondary BF separation process on the control sample in the second container 12. Next, the R4 reagent dispensing unit 134 and the R5 reagent dispensing unit 135 dispense the R4 reagent containing the buffer solution and the R5 reagent containing the substrate into the second container 12.

(Embodiment 3)
The example of FIG. 14 is basically the same as the example shown in FIG. 13, but the solid phase carrier 82 to be bound to the labeling substance 83 is different. In the example of FIG. 14, the reagent dispensing order is different from the example of FIG.

Also in the example of FIG. 14, the state confirmation method includes a sample containing a test substance 81 having a known concentration in the first container 11 before the step of transferring the liquid phase in the first container 11 to the second container 12. A step of contacting a labeling substance 83 that binds to the test substance 81, a capture substance 86 that binds to the test substance 81, and a solid phase carrier 82 that binds to the capture substance 86;

In the example of FIG. 14, first, the r4 reagent containing the capture substance 86 is dispensed into the first container 11. Next, a quality control sample containing a test substance 81 having a known concentration is dispensed. Next, the r1 reagent containing the labeling substance 83 is dispensed into the first container 11. Then, the r7 reagent containing the second solid phase carrier 82b is dispensed. As a result, the immune complex 84 including the test substance 81, the labeling substance 83, and the capture substance 86 is bound to the second solid phase carrier 82b. Reagent dispensing is performed by the reagent dispensing unit 130, and dispensing of the quality control sample is performed by the sample dispensing unit 120.

As described above, in the example of FIG. 14, the management sample 20 includes the second solid phase carrier 82b to be dispensed into the second container 12 after the complex transfer process at the time of measurement of the specimen. That is, the management sample 20 includes the second solid phase carrier 82b that is a magnetic particle included in the r7 reagent. Thereby, the solid phase carrier 82 to which the labeling substance 21 is bound is formed in the first container 11 by using the second solid phase carrier 82b dispensed when the specimen including the test substance 81 is actually measured. be able to. That is, since it is not necessary to separately prepare a dedicated solid phase carrier 22 for confirming the state of the immunoassay device 100, the convenience of the immunoassay device 100 can be improved.

Next, primary BF separation processing is performed by the BF separation unit 170. In the state confirmation method, before the step of transferring the liquid phase in the first container 11 to the second container 12, the solid phase carrier 82 bound to the immune complex 84 including the test substance 81, the labeling substance 83, and the capture substance 86. And a step of performing BF separation for separating the liquid phase.

After the primary BF separation process, the liquid phase 25 not containing the free reagent 85 is dispensed into the first container 11. In the state confirmation method, after the step of performing BF separation, before the step of transferring the liquid phase in the first container 11 to the second container 12, the liquid phase 25 that does not contain the free reagent 85 is dispensed into the first container 11. The process of carrying out is provided. The liquid phase 25 not containing the free reagent 85 is, for example, a buffer solution. Since the liquid phase 25 does not contain the free reagent 85, the labeling substance 83 and the solid phase carrier 82 remain bonded.

Next, a complex transfer process is performed. During the step (a) of transferring the liquid phase in the first container 11 containing the control sample 20 to the second container 12, the second solid phase carrier 82 b in the first container 11 is Magnetized in the container 11. The liquid phase in the first container 11 is sucked and dispensed into the second container 12 by the suction pipe 51 in the state of being magnetized. The second solid phase carrier 82b is left magnetized in the first container 11. Since the immune complex 84 remains bound to the second solid phase carrier 82b, the liquid phase not containing the labeling substance 83 is transferred to the second container 12.

Next, the reagent dispensing unit 130 dispenses the r7 reagent including the second solid phase carrier 82b into the second container 12 to which the liquid phase has been transferred. Thus, in the example of FIG. 14, the r7 reagent including the second solid phase carrier 82b is dispensed before and after the complex transfer treatment. Then, the BF separation unit 170 performs a secondary BF separation process on the control sample in the second container 12. Next, the R4 reagent dispensing unit 134 and the R5 reagent dispensing unit 135 dispense the R4 reagent containing the buffer solution and the R5 reagent containing the substrate into the second container 12.

Next, a step (b) of detecting the labeling substance 83 in the second container 12 to which the liquid phase has been transferred is performed by the detection unit 60. By the step (b), a detection value indicating the abundance of the labeling substance 83 is acquired.

As described above, in the example shown in FIGS. 13 and 14, the mechanism unit 50 includes the sample dispensing unit 120 for dispensing the sample including the test substance 81 and the labeling reagent (r1 reagent including the labeling substance 83). And a reagent dispensing unit 130 for dispensing a solid phase reagent (r5 reagent or r7 reagent) including the solid phase carrier 82, and the management sample 20 is prepared in the first container 11. It is configured. Thereby, even if the user or service staff of the immunoassay apparatus 100 does not prepare the management sample 20 in the first container 11 in advance, the management sample 20 is easily accommodated in the first container 11 by the immunoassay apparatus 100. Can be made.

In the example shown in FIGS. 13 and 14, the reagent dispensing unit 130 provides the first container 11 with a free reagent 85 that releases the immune complex 84 from the solid phase carrier 82 when measuring the sample. When determining an abnormality in the process of dispensing and transferring the liquid phase in the first container 11 to the second container 12, the liquid phase 25 not containing the free reagent 85 is dispensed instead of the free reagent 85. Thereby, in place of the free reagent 85 that is dispensed when the specimen containing the test substance 81 is actually measured, the liquid phase that does not contain the free reagent 85 is dispensed, so that the free reagent 85 is measured during the actual specimen measurement. The state of the immunoassay device 100 can be confirmed by the same operation as that of dispensing. Even in this case, since the binding of the labeling substance 21 to the solid phase carrier 22 is not eliminated, the magnetic flux collecting function of the solid phase carrier 22 can be appropriately confirmed.

In the example shown in FIGS. 13 and 14, the mechanism unit 50 includes a BF separation unit that separates the solid phase carrier 82 bound to the immune complex 84 including the test substance 81 and the labeling substance 83 from the liquid phase. 170, the BF separation unit 170 performs BF separation on the control sample 20 in the first container 11. That is, the BF separation unit 170 performs the primary BF separation process. Thereby, the labeling substance 83 in the liquid phase that has not been bonded to the solid phase carrier 82 can be separated from the solid phase carrier 82 and eliminated by BF separation. Therefore, in the step of transferring the liquid phase in the first container 11 to the second container 12, it is possible to prevent the labeling substance 83 from being mixed in the liquid phase, so that the detection of the labeling substance 83 for confirming the state of the immunoassay device 100 is detected. Accuracy can be improved.

<Process for judging magnetism collecting function>
As described above, in the examples of Embodiments 1 to 3 shown in FIGS. 12 to 14, the labeling substance 21 is bound to the solid phase carrier 22 in the control sample 20, and the magnetism is collected in the first container 11. Thus, the labeling substance 21 is left in the first container 11 together with the solid phase carrier 22 after the step (a) of transferring the liquid phase to the second container 12. Therefore, in the examples of Embodiments 1 to 3, when the magnetic flux collecting function is normal, as a detection result in the step (b), measurement is performed on a blank sample that does not substantially contain the labeling substance 21. A detection value of the same level as is acquired. More precisely, even if the magnetism collecting function is normal, not all the solid phase carriers 22 in the first container 11 can be magnetized, so that the solid phase carriers 22 in the first container 11 are very small. A part is carried over to the 2nd container 12 with a liquid phase. Therefore, the allowable carryover amount of the solid phase carrier 22 in the specification is set for the magnetism collecting function. The allowable carryover amount is set as a variation in a range that does not affect the detection result. As a reference value V1 (see FIG. 11) for determining whether or not the magnetic flux collecting function is normal, a value obtained by adding an upper limit range of the allowable carryover amount to an expected detection value when a blank sample is measured. Is set. When the acquired detection value is lower than the reference value V1, it is determined that the magnetism collecting function is normal.

On the other hand, when the magnetism collecting function is abnormal, for example, when the solid phase carrier 22 cannot be magnetized and is dispersed in the liquid phase of the first container 11, the solid substance bonded to the labeling substance 21 in step (a) The phase carrier 22 is transferred to the second container 12 together with the liquid phase. As a result, in the examples of the first to third embodiments, when the magnetic flux collecting function is abnormal, the detection value significantly increases as the detection result in the step (b) exceeds the upper limit of the allowable carryover amount. Therefore, when the acquired detection value becomes the reference value V1 or more, it is determined that the magnetism collecting function is abnormal.

Example 1
FIG. 15 shows experimental conditions of Example 1 performed for confirming the effect of the magnetic flux collecting function determination process, and FIG. 16 shows experimental results of Example 1. FIG. In Example 1, the procedure of Embodiment 3 shown in FIG.

In the comparative example of FIG. 15, in order to contrast with Example 1, each processing step at the time of measurement shown in FIG. 4 is listed. As shown in FIG. 15, in Example 1, the HISCL HBsAg R1 reagent (manufactured by Sysmex) was used as the r4 reagent, the HISCL HBsAg calibrator C4 (manufactured by Sysmex) was used as the quality control sample, and the HISCL HBsAg R3 reagent was used as the r1 reagent. (Manufactured by Sysmex Corporation), HICL CLB4 reagent (manufactured by Sysmex Corporation) as the R7 reagent, HISCL HBsAg R2 reagent (manufactured by Sysmex Corporation) as the r7 reagent, HICL CL wash solution (manufactured by Sysmex Corporation) as the liquid phase 25 not containing the free reagent 85 HISCL R5 reagent (manufactured by Sysmex Corporation) was used as the R5 reagent. The amount dispensed is as shown in FIG.

As shown in FIG. 16, in Example 1, in order to compare the normal state and the abnormal state of the magnetic flux collection function, when magnetic flux collection is performed (with magnets), magnetic flux collection is performed using the magnetic force source 52. The experiment was performed in the case of no (no magnet) and the procedure of FIG. Each experiment was performed 20 times, and the average value, standard deviation (SD), coefficient of variation (C.V.), maximum value, minimum value, and range (difference between the maximum value and the minimum value) of the detected values were obtained. . The detection value is a count number of photons acquired by the photodetector 61 of the detection unit 60.

From FIG. 16, the detection value is acquired in the range of about 1000 counts to about 2000 counts in normal (with magnets), and the detection value is acquired in the range of about 16 million counts to about 19 million counts in the abnormal (without magnets). It was done. The detection value of normal (with a magnet) is approximately the same as the measurement value when a blank sample is measured. The detected value of the abnormality (no magnet) substantially matches the detected value when the quality control sample is measured in the processing step shown in FIG. There is a difference of about 10,000 times in the detected value between normal (with magnet) and abnormal (without magnet). From Example 1, it is possible to clearly distinguish the normal range and the abnormal range of the detected value even in consideration of the carry-over amount of the magnetic particles in the normal state and the fluctuation of the detected value due to the secular change of the immunoassay device. It was confirmed that it was possible to determine normality and abnormality of the magnetic flux collecting function in the complex transition process by appropriately setting the reference value V1 for determination.

<Confirmation of dispensing function>
With reference to FIG. 18 and FIG. 19, the example which confirms the dispensing function of the immunoassay apparatus 100 is shown. In the example of FIGS. 18 and 19, the management sample 20 includes a solution in which the labeling substance 21 is dissolved. That is, in the control sample 20, the labeling substance 21 does not bind to the solid phase but exists as a liquid phase. In this case, since the labeling substance 21 exists in the first container 11 as the liquid phase, for example, when the detection value of the labeling substance 21 does not increase, the first container 11 is transferred when the liquid phase is transferred to the second container 12. The liquid phase may not be transferred properly. Therefore, it can be easily confirmed whether or not the function of transferring the liquid phase in the first container 11 to the second container 12 in the complex transfer process is normal.

Specifically, in the example of FIGS. 18 and 19, in the step (a) of transferring the liquid phase in the first container 11 to the second container 12, the liquid phase in the first container 11 is sucked and sucked liquid. A dispensing process for discharging the phase to the second container 12 is included. The state confirmation method includes a step of determining an abnormality in the dispensing function based on the detection result of the labeling substance 21 in the second container 12. Thereby, based on the detected value of the labeling substance 21, it can be easily determined whether the dispensing function for sucking the liquid phase and discharging the liquid phase is normal or abnormal.

The determination of the dispensing function determines that there is an abnormality in the dispensing function when the detected value of the labeling substance 21 is below the reference value V2, as shown in FIG. Thereby, when the detection value of the labeling substance 21 is lower than the reference value V2, it is highly likely that the liquid phase containing the labeling substance 21 is not properly transferred from the first container 11 to the second container 12. The dispensing function can be easily determined by comparing the value with the reference value V2.

The reference value V2 for determining the dispensing function is a value set as an allowable limit of the expected detection value of the labeling substance 21 included in the control sample 20. Thereby, even if it performs the process of moving the liquid phase containing the labeling substance 21 to the 2nd container 12, when a detected value is less than the reference value V2, it can determine easily that a dispensing function is abnormal. Therefore, the reliability of the measurement result can be ensured by confirming that the detected value exceeds the reference value V2.

(Embodiment 4)
In the example of FIG. 18, a labeling substance different from the labeling substance 83 used at the time of measuring the sample is used as the labeling substance 21 for confirming the dispensing function. That is, in the example of FIG. 18, the labeling substance 21 for confirming the dispensing function is a quality control reagent prepared separately from the labeling substance 83 included in the r1 reagent when measuring the sample. It is included.

In the example of FIG. 18, the management sample 20 in which the labeling substance 21 is dissolved in the liquid phase is accommodated in the first container 11. The labeling substance 21 is accommodated in the first container 11 as a liquid phase without being bound to the solid phase. The management sample 20 is dispensed into the first container 11 by the mechanism unit 50. For example, the control sample 20 is set in the sample transport unit 190 while being stored in a predetermined control sample container in the same manner as the sample container 191, and is dispensed into the first container 11 by the sample dispensing unit 120. Further, for example, the management sample 20 is set in the reagent storage 150 while being accommodated in a predetermined management sample container in the same manner as the reagent container 155, and is dispensed into the first container 11 by the reagent dispensing unit 130.

In the example of FIG. 18, the dispensing of the r1 reagent, the r4 reagent, the r5 reagent, and the r6 reagent is skipped or dispensed with a buffer solution or the like. In the example of FIG. 18, the primary BF separation process is skipped. When the alternative liquid is dispensed, a primary BF separation process may be performed.

Next, in the step (a) of transferring the liquid phase in the first container 11 in which the control sample 20 is accommodated to the second container 12, the liquid phase in the first container 11 is sucked by the suction tube 51 and the second phase is moved. It is dispensed into the container 12.

In the example of FIG. 18, the dispensing of the r7 reagent after the complex transfer treatment is skipped or dispensed with a buffer solution or a washing solution. In the example of FIG. 18, the secondary BF separation process is skipped. When the labeling substance 21 is an enzyme label, a substrate corresponding to the enzyme is dispensed as the R5 reagent.

In the example of FIG. 18, an example in which the management sample 20 is dispensed at the r6 reagent dispensing timing shown in FIG. 4 is shown, but at any timing the management sample 20 is before the complex transfer process. You may dispense.

(Embodiment 5)
The example of FIG. 19 is an example in which the same labeling substance as the labeling substance 83 used when measuring the specimen is used as the labeling substance 21 for confirming the dispensing function. That is, in the example of FIG. 19, the management sample 20 includes the labeling substance 83 of the r1 reagent dispensed into the first container 11 when the specimen is measured. Accordingly, whether or not the function of transferring the liquid phase in the first container 11 to the second container 12 is normal using the labeling substance 83 dispensed when the specimen including the test substance 81 is actually measured. I can confirm. That is, it is not necessary to separately prepare a dedicated labeling substance 83 for confirming the state of the immunoassay device 100, so that the convenience of the immunoassay device 100 can be improved.

In the example of FIG. 19, a management sample in which the labeling substance 83 is dissolved in the liquid phase is accommodated in the first container 11. The labeling substance 83 is accommodated in the first container 11 as a liquid phase without being bound to the solid phase. The r1 reagent containing the labeling substance 83 is aspirated from the reagent container 155 set in the reagent container 150 by the reagent dispensing unit 130 and dispensed into the first container 11.

19 shows an example in which the buffer solution is dispensed as an alternative liquid at the dispensing timing of the r1 reagent and the specimen shown in FIG. Dispensing of the r4 reagent, r5 reagent, and r6 reagent is skipped. Dispensing of the buffer solution may be skipped, and in this case, the control sample 20 may be the r1 reagent itself. The example of FIG. 19 shows an example in which the r1 reagent is dispensed at the r6 reagent dispensing timing shown in FIG. 4, but at any timing the r1 reagent is dispensed before the complex transfer treatment. May be. After dispensing the r1 reagent, the primary BF separation process is skipped.

In the example of FIG. 19, the dispensing of the r7 reagent after the complex transfer treatment is skipped or dispensed with a buffer solution or the like. In the example of FIG. 19, the secondary BF separation process is skipped. In the example of FIG. 19, the R4 reagent and the R5 reagent are dispensed into the second container 12.

(Example 2)
FIG. 20 shows the experimental conditions of Example 2 performed to confirm whether the dispensing function can be determined, and FIG. 21 shows the experimental results of Example 2. In Example 2, the procedure of the fifth embodiment shown in FIG. For comparison with Example 2, the comparative example of FIG. 20 lists each processing step at the time of measurement shown in FIG.

As shown in FIG. 20, in Example 2, the HISCL HBsAg R3 reagent (manufactured by Sysmex Corporation) was dispensed as the r1 reagent containing the labeling substance 83 at the dispensing timing of the r6 reagent at the time of measurement. The secondary BF separation process after the complex transfer process was skipped. HICL CL4 reagent (manufactured by Sysmex) was used as the R4 reagent, and HISCL R5 reagent (manufactured by Sysmex) was used as the R5 reagent.

As shown in FIG. 21, in Example 2, the dispensing amount in the step (a) of transferring the liquid phase of the first container 11 to the second container 12 is different in order to compare the normal state and the abnormal state of the dispensing function. The experiment was conducted under the conditions described above. In Example 2, the dispensing amount in the normal state was set to 20 [μL], the dispensing amount in the abnormal state was set to 10 [μL], and the experiment was performed according to the procedure of FIG. Each experiment was performed 20 times, and the average value, standard deviation (SD), coefficient of variation (C.V.), maximum value, minimum value, and range (difference between the maximum value and the minimum value) of the detected values were obtained. . The detection value is a count number of photons acquired by the photodetector 61 of the detection unit 60.

From FIG. 21, in normal (20 μL), detection values are acquired in the range of about 130 million counts to about 150,000,000, and in abnormal (10 μL), detection values are acquired in the range of about 80 million counts to about 90 million counts. It was done. There is a difference of 1.5 times or more in the detected value between normal (20 μL) and abnormal (10 μL). From Example 2, it can be seen that when the dispensing amount of the liquid phase into the second container 12 varies due to an abnormality, the detected value varies according to the dispensing amount. Therefore, by setting in advance a reference value V2 that defines the allowable range of the expected detection value of the labeling substance 83 contained in the control sample 20 from the set value of the liquid phase dispensing amount, It was confirmed that the order function can be judged normal or abnormal. For example, even if a range of (average value ± 2SD) is set as the allowable range, the normal state and the abnormal state can be sufficiently determined.

Note that the immunoassay apparatus 100 may be capable of arbitrarily setting the amount of liquid phase dispensed in the complex transfer process within a variable range. For example, in the immunoassay device 100, the user can arbitrarily set the dispensing amount of the liquid phase in the range of 10 μL to 80 μL. Also in this case, it can be seen from Example 2 that the detected value varies depending on the dispensed amount, and therefore, the reference value V2 that defines the allowable range of the predicted detected value corresponding to the preset dispensed amount of the liquid phase. Can be obtained in advance. The determination unit 70 acquires the set value of the liquid phase dispensing amount, selects the reference value V2 corresponding to the set value, and compares it with the detected value, so that the dispensing function in the complex transfer process is normal. Can determine if it is abnormal.

(Modification of status check method)
In the state confirmation method, in addition to the above steps for confirming the magnetism collecting function and the dispensing function, another state confirmation step may be performed.

Specifically, the state confirmation method includes a sample containing a test substance 81 having a known concentration or a sample not containing the test substance 81 in the first container 11, a labeling substance 21 that binds to the test substance 81, a test The step of contacting the capture substance 86 that binds to the substance 81 and the solid phase carrier 22 that binds to the capture substance 86 and the state confirmation method include an immune complex including the labeling substance 21 and the capture substance 86 in the first container 11. After dispensing the free reagent 85 that liberates 84 from the solid phase carrier 22, the step of transferring the liquid phase in the first container 11 to the second container 12 and the labeling substance 21 present in the second container 12 are detected. You may further provide a process.

That is, instead of the specimen shown in FIG. 4, by using a quality control sample containing a test substance 81 having a known concentration or a sample not containing the test substance 81, the same operation as the measurement operation for a normal specimen is performed. The status may be checked. By measuring the control sample containing the test substance 81 having a known concentration, it is confirmed that the detection value obtained in the step (b) falls within the range of the reference value expected from the control sample having the known concentration. Further, the measurement value of the control sample that does not include the test substance 81 causes the detection value obtained in the step (b) to be within the range of reference values expected from the control sample that does not include the test substance 81 (blank sample). Confirmed to fit. Thereby, it is confirmed that the measurement accuracy of the immunoassay using the complex transfer treatment is normal.

(Explanation of the operation of the immunoassay device)
Next, the operation of the immunoassay apparatus 100 shown in FIG. 5 will be described with reference to FIG. In the following description, each step of the operation of the immunoassay apparatus 100 is referred to FIG. 22, and each part of the immunoassay apparatus 100 is referred to FIGS. For the status confirmation processing, refer to FIG. 11 to FIG. 14 and FIG. 17 to FIG. Operation control of the following steps is performed by the analysis unit 110 of the immunoassay device 100.

In step S1, the analysis unit 110 determines whether or not to check the state of the immunoassay device 100. The state confirmation is performed, for example, when a user inputs a state confirmation execution command via an input device (not shown) such as a touch panel or a mouse. The state confirmation is executed, for example, when a predetermined state confirmation execution timing comes. The execution timing of the status check is at least before the start of the first measurement on the first day, or at the first device startup on the first day. The execution timing of the state confirmation can be arbitrarily set in advance by the user.

If the status of the immunoassay apparatus 100 is not checked, the analysis unit 110 determines whether or not to perform immunoassay of the sample in step S2. The analysis unit 110 determines whether or not to perform immunoassay based on whether or not the measurement order is registered in the host computer. When the measurement order is registered, in step S3, the analysis unit 110 causes the mechanism unit 50 to start measurement processing. The immunoassay is performed according to the procedure shown in FIG. When the detection result of the detection unit 60 is acquired, the analysis unit 110 compares the detection value with the calibration curve and obtains the abundance of the test substance 81. If the measurement order is not registered in step S2, the process returns to step S1 and waits until the start timing for state confirmation or immunoassay.

In step S1, when the state of the immunoassay apparatus 100 is confirmed, the process proceeds to step S4. In steps S4 to S7, the analysis unit 110 causes the mechanism unit 50 to perform the state confirmation operation shown in FIGS. Here, the dispensing and the primary BF separation process performed before the complex transfer process are described as the preparation process of the control sample in step S4 for convenience. In addition, the dispensing and the secondary BF separation process performed after the complex transfer process are described as post-processing of step S6 for convenience.

Here, an example is shown in which both the confirmation of the magnetism collecting function and the confirmation of the dispensing function of the immunoassay apparatus 100 are performed. In this case, the analysis unit 110 includes the first container 11 containing the management sample 20 including the

solid phase carrier

22 or 82 to which the

labeling substance

21 or 83 is bound, as shown in FIGS. As shown in FIG. 19, the first container 11 containing the control sample 20 containing the solution in which the

labeling substance

21 or 83 is dissolved is prepared. Then, the magnetic flux collecting function is confirmed using the first container 11 containing the control sample containing the

solid phase carrier

22 or 82 to which the

labeling substance

21 or 83 is bound, and the solution in which the

labeling substance

21 or 83 is dissolved is obtained. The dispensing function is confirmed using the first container 11 containing the management sample 20 including the control sample 20.

In step S5, the analysis unit 110 causes the mechanism unit 50 to perform a complex transfer process. That is, the analysis unit 110 causes the mechanism unit 50 to perform the step (a) of transferring the liquid phase in the first container 11 containing the management sample 20 including the labeling substance 21 to the second container 12.

In step S7, the analysis unit 110 causes the detection unit 60 to perform the step (b) of detecting the labeling substance 21 in the second container 12 to which the liquid phase has been transferred. As a result of step S7, the measurement result of the control sample is acquired.

In step S8, the determination unit 70 determines a function for performing the complex transfer process. The determination unit 70 determines whether the magnetic collection function is normal or abnormal based on whether the detection value of the detection unit 60 is lower than the reference value V1 (see FIG. 11). . For the dispensing function, the determination unit 70 determines whether the dispensing function is normal or abnormal based on whether the detection value of the detection unit 60 exceeds the reference value V2 (see FIG. 17). .

When it is determined that the magnetism collecting function and the dispensing function are normal, the analysis unit 110 waits for immunoassay of the specimen. When it is determined that the magnetism collecting function or the dispensing function is abnormal, the analysis unit 110 has an abnormality in the function of performing the complex transfer process of the immunoassay device 100 by, for example, displaying a message on a display device (not shown). This is notified to the user. At this time, the analysis unit 110 may stop the function of performing the immunoassay of the specimen until the state confirmation is performed again and it is determined that the magnetism collecting function and the dispensing function are normal.

In addition, it should be thought that embodiment disclosed this time is an illustration and restrictive at no points. The scope of the present invention is shown not by the above description of the embodiments but by the scope of claims for patent, and further includes all modifications within the meaning and scope equivalent to the scope of claims for patent.

The present invention can be suitably used for confirmation of the state of an immunoassay device using, for example, an immune complex transfer method.

11: first container, 12: second container, 20: control sample, 21: labeling substance, 22: solid phase carrier, 23: third solid phase carrier, 50: mechanism, 51: suction tube, 52: magnetic source 60: detection unit, 70: determination unit, 81: test substance, 82: solid phase carrier, 82a: first solid phase carrier, 82b: second solid phase carrier, 83: labeling substance, 84: immune complex, 85: free reagent, 86: capture substance, 100: immunoassay device, 110: analysis unit, 120: sample dispensing unit, 121: suction tube, 130: reagent dispensing unit, 131: suction tube, 170: BF separation unit , V1: reference value, V2: reference value

Claims

In a first container that contains an immune complex that includes a test substance and a labeling substance in a sample and is supported on a solid phase carrier, the immune complex is released from the solid phase carrier, A method for confirming the state of an immunoassay device for performing a complex transfer process for transferring a liquid phase to a second container,
Transferring the liquid phase in the first container containing the control sample containing at least the labeling substance to the second container;
Detecting the labeling substance in the second container to which the liquid phase has been transferred;
Determining the state of the immunoassay device that performs the complex transfer process based on the detection result of the labeling substance in the second container.
The method for confirming the state of the immunoassay device according to claim 1, further comprising a step of determining an abnormality in the complex transfer process based on a detection result of the labeling substance in the second container.
The step of determining an abnormality in the complex transfer process includes at least one of a function of collecting the solid phase carrier in the first container and a function of transferring the liquid phase in the first container to the second container. The method for confirming the state of the immunoassay device according to claim 2, comprising determining whether or not the abnormality is present.
The state of the immunoassay device according to claim 2, wherein the abnormality is determined based on whether a detected value of the labeling substance in the second container exceeds a reference value or the detected value is lower than a reference value. Confirmation method.
The immunity according to claim 1, further comprising the step of dispensing the management sample into the first container by the immunoassay device before the step of transferring the liquid phase in the first container to the second container. How to check the status of the measuring device.
The management sample includes the solid phase carrier to which the labeling substance is bound,
2. The immunoassay device according to claim 1, wherein in the step of transferring the liquid phase in the first container to the second container, the solid phase carrier to which the labeling substance is bound is collected in the first container. Status check method.
The solid support is magnetic particles,
The process of collecting the solid phase carrier includes a process of collecting the magnetic particles by a magnetic source,
The method for confirming the state of the immunoassay device according to claim 6, further comprising a step of determining an abnormality of the magnetic flux collecting function by the magnetic force source based on a detection result of the labeling substance in the second container.
The method of confirming the state of the immunoassay device according to claim 7, wherein when the detected value of the labeling substance exceeds a reference value, it is determined that there is an abnormality in the magnetic flux collecting function by the magnetic force source.
The method for confirming the state of the immunoassay device according to claim 8, wherein the reference value is an expected detection value of the labeling substance corresponding to an allowable upper limit amount of the magnetic particles transferred to the second container without being collected. .
Before the step of transferring the liquid phase in the first container to the second container, the sample containing the test substance having a known concentration in the first container, the labeling substance that binds to the test substance, The method for confirming the state of the immunoassay device according to claim 6, further comprising a step of contacting a capture substance that binds to a test substance and the solid phase carrier that binds to the capture substance.
The method for confirming a state of an immunoassay device according to claim 10, wherein the management sample includes a first solid phase carrier that is dispensed into the first container before complex transfer treatment at the time of measurement of a specimen.
The method for confirming the state of the immunoassay device according to claim 10, wherein the management sample includes a second solid phase carrier that is dispensed into the second container after complex transfer treatment at the time of measurement of the specimen.
Before the step of transferring the liquid phase in the first container to the second container, the solid phase carrier bound to the immune complex including the test substance, the labeling substance, and the capture substance, and a liquid phase The method for confirming the state of the immunoassay device according to claim 10, further comprising a step of performing BF separation for separating the immunity.
In the complex transfer treatment, a free reagent is dispensed into the first container, whereby the immune complex is released from the solid phase carrier,
After the step of performing BF separation, before the step of transferring the liquid phase in the first container to the second container, further comprising the step of dispensing the liquid phase not containing the free reagent into the first container. The method for confirming the state of the immunoassay device according to claim 13.
The method for confirming the state of the immunoassay device according to claim 6, wherein the management sample includes a third solid phase carrier that is bound to the labeling substance without being bound to the test substance.
The method for confirming the state of the immunoassay device according to claim 1, wherein the control sample includes a solution in which the labeling substance is dissolved.
The step of transferring the liquid phase in the first container to the second container includes a dispensing process for sucking the liquid phase in the first container and discharging the sucked liquid phase to the second container,
The method of confirming the state of the immunoassay device according to claim 16, further comprising a step of determining an abnormality in a dispensing function based on a detection result of the labeling substance in the second container.
The method for confirming the state of the immunoassay device according to claim 17, wherein when the detection value of the labeling substance is below a reference value, it is determined that there is an abnormality in the dispensing function.
19. The method according to claim 18, wherein the reference value is a value set as an allowable limit of an expected detection value of the labeling substance contained in the control sample.
The method for confirming the state of the immunoassay device according to claim 16, wherein the control sample includes the labeling substance dispensed into the first container at the time of measurement of a specimen.
The labeling substance contained in the control sample is an enzyme;
The step of detecting the labeling substance in the second container is performed by adding a substrate of the enzyme to the second container and measuring a signal generated from a reaction product generated by an enzyme reaction. The state confirmation method of the immunoassay apparatus of description.
In the first container, a sample containing the test substance having a known concentration or a sample not containing the test substance, the labeling substance that binds to the test substance, a capture substance that binds to the test substance, and the Contacting the solid phase carrier that binds to the capture substance;
After dispensing a free reagent for releasing the immune complex containing the labeling substance and the capture substance from the solid phase carrier into the first container, the liquid phase in the first container is transferred to the second container. The method of confirming the state of the immunoassay device according to claim 1, further comprising a step of transferring, and a step of detecting the labeling substance present in the second container.
In a first container that contains an immune complex that includes a test substance and a labeling substance in a sample and is supported on a solid phase carrier, the immune complex is released from the solid phase carrier, An immunoassay device for performing a complex transfer process for transferring a liquid phase to a second container,
A mechanism for performing a process of transferring the liquid phase in the first container containing the control sample containing at least the labeling substance to the second container;
A detection unit for detecting the labeling substance in the second container to which the liquid phase has been transferred;
An immunoassay device comprising: a determination unit that determines an abnormality in the complex transfer process based on a detection result of the detection unit.
The determination unit is at least one of a function of collecting the solid phase carrier in the first container by the mechanism unit and a function of transferring the liquid phase in the first container to the second container by the mechanism unit. The immunoassay device according to claim 23, wherein abnormality is determined.
The management sample includes magnetic particles to which the labeling substance is bound,
The mechanism unit includes a magnetic force source that collects the magnetic particles to which the labeling substance is bound in the first container,
25. The immunoassay device according to claim 24, wherein the determination unit determines abnormality of a magnetic flux collecting function by the magnetic source.
The control sample includes a solution in which the labeling substance is dissolved,
The mechanism portion includes a suction pipe that sucks the liquid phase in the first container and discharges the sucked liquid phase to the second container;
The immunoassay device according to claim 24, wherein the determination unit determines an abnormality in a dispensing function by the suction tube.
The mechanism unit is a reagent for dispensing a sample dispensing unit for dispensing a sample containing the test substance, a labeling reagent containing the labeling substance, and a solid phase reagent containing the solid phase carrier. 24. The immunoassay device according to claim 23, comprising a dispensing unit, and configured to prepare the management sample in the first container.
The reagent dispensing unit is
When measuring the specimen, a free reagent for releasing the immune complex from the solid phase carrier is dispensed into the first container,
28. When determining an abnormality in the process of transferring the liquid phase in the first container to the second container, the liquid phase not containing the free reagent is dispensed instead of the free reagent. Immunoassay device.
The mechanism part includes a BF separation part for separating the solid phase carrier bound to the test substance, the immune complex containing the labeling substance, and a liquid phase,
28. The immunoassay device according to claim 27, wherein the BF separation unit performs BF separation on the management sample in the first container.