JP2019082477A - Analytical curve generation method and autoanalyzer - Google Patents

Abstract

To measure a detection object with high sensitivity and high concentration, that is, to achieve a wide dynamic range with high sensitivity in measurement using an insoluble carrier having an antibody or an antigen immobilized thereon.SOLUTION: An analytical curve generation method includes generating an analytical curve on the basis of reaction curves C1 to C8 obtained from photometric results of a plurality of standard samples corresponding to different concentration including a detection object having known concentration. The analytical curve generation means includes generating the analytical curve on the basis of the photometric results of the plurality of standard samples obtained at different photometric timings.SELECTED DRAWING: Figure 5

Description

  Embodiments of the present invention relate to a calibration curve generation method and an automatic analyzer.

  The automatic analyzer adds reagents corresponding to various test items to a test sample such as blood, and causes the reagent to react with specific components contained in the test sample. The automatic analyzer analyzes the components of the test sample corresponding to the test item by optically measuring this reaction solution, for example.

  In an automatic analyzer, a technique for achieving high sensitivity of measurement using a latex agglutination method is known. In the latex aggregation method, a reagent in which an antibody is bound to the surface of latex particles, which is an insoluble carrier, is added to a sample to form a reaction solution. In the reaction solution, the antigen contained in the sample is bound to the antibody on the surface of the latex particle to aggregate the latex particle. Then, in the process of aggregation of the latex particles, the reaction solution is irradiated with light, and the transmitted light passing through the reaction solution or the intensity of the scattered light scattered in the reaction solution is measured to detect the sample contained in the sample. Measure the amount of component of interest.

  In the case of the agglutination method, if the particle size is increased, the reaction area per fixed time decreases and the antigen-antibody reaction decreases, but the scattering increases and even a slight aggregation can be detected. Conversely, if the particle size is small, the reaction area per fixed time will be large and the reaction will be large, but the scattering is small and detection is difficult. In general, the particle concentration is limited when a large diameter is used so that high sensitivity can be measured by the latex agglutination method. For example, when the particle concentration is higher than or equal to a certain level, the transmitted light is reduced, which leads to a decrease in detectability. In addition, when the concentration of the substance to be measured is high, the clumps become too large and self-sedimentation occurs, causing the clumps to deviate from the optical path and the absorbance to decrease. Therefore, when using large diameter particles, it is difficult to secure a dynamic range.

  On the other hand, when particles of small diameter are used, the particle concentration can be increased, so that measurement can be performed up to the high concentration region of the measurement object. However, although self-sedimentation due to aggregation hardly occurs and a wide dynamic range can be secured, detection sensitivity is degraded. The current latex agglutination method reagent achieves high sensitivity and wide range by using particles of two or more kinds of particle sizes of a large particle size with high sensitivity and a small particle size capable of obtaining a dynamic range. However, since the advantages of both have been gradually eliminated, the inherent performance of single-size particles has been reduced.

JP-A-2014-192963 Patent No. 6104746 specification

  The problem to be solved by the invention is that in the measurement using the insoluble carrier on which the antibody or antigen is immobilized, the detection object can be measured with high sensitivity and high concentration, that is, high sensitivity and wide dynamic range are realized. It is.

  According to an embodiment, the calibration curve generation method generates a calibration curve based on photometric results of a plurality of standard samples corresponding to different concentrations, including detection targets of known concentrations. The calibration curve generation method generates the calibration curve based on photometric results of a plurality of standard samples acquired at different photometric timings.

FIG. 1 is a block diagram showing a functional configuration of the automatic analyzer according to the first embodiment. FIG. 2 is a diagram showing a photometric timing line and photometric timing according to the first embodiment. FIG. 3 is a diagram showing the configuration of the analysis mechanism shown in FIG. FIG. 4 is a diagram showing measured values and response curves for standard samples. FIG. 5 is a diagram showing a photometric timing line set based on the response curve shown in FIG. 4 and the photometric timing. FIG. 6 is a diagram showing a photometric timing line set based on the response curve shown in FIG. 4 and the photometric timing. FIG. 7 is a diagram showing a photometric timing line set based on the measurement value and the photometric timing. FIG. 8 shows a standard calibration curve generated based on the photometric timing shown in FIG. FIG. 9 is a diagram showing a light measurement timing line and a light measurement timing when two light measurement timing lines are set. FIG. 10 is a diagram showing a standard calibration curve generated based on the photometric timing shown in FIG. FIG. 11 is a diagram showing a response curve, a photometric timing line, and a test data approximate curve. FIG. 12 is a diagram showing corrected calibration data corresponding to FIG. FIG. 13 is a diagram illustrating an example of a method of acquiring a standard calibration curve according to the second embodiment. FIG. 14 shows a standard calibration curve obtained by the method of FIG. FIG. 15 is a diagram illustrating another example of the method of acquiring a standard calibration curve. FIG. 16 is a diagram showing a standard calibration curve acquired by the method of FIG. FIG. 17 is a diagram illustrating another example of the method of acquiring a standard calibration curve. FIG. 18 is a diagram showing a standard calibration curve acquired by the method of FIG. FIG. 19 is a diagram illustrating another example of the method of acquiring a standard calibration curve. FIG. 20 is a diagram showing a part of a standard calibration curve acquired by the method of FIG. FIG. 21 is a diagram showing a part of a standard calibration curve acquired by the method of FIG. FIG. 22 is a diagram showing a response curve, a photometric timing line, and a test data approximate curve. FIG. 23 is a diagram showing corrected calibration data corresponding to FIG. FIG. 24 is a diagram showing a response curve, a photometric timing line, and a test data approximate curve. FIG. 25 is a diagram showing corrected calibration data corresponding to FIG.

  Embodiments will be described below with reference to the drawings.

First Embodiment
(Automatic analyzer)
FIG. 1 is a block diagram showing an example of the functional configuration of the automatic analyzer 1 according to the first embodiment. The automatic analyzer 1 shown in FIG. 1 includes an analysis mechanism 2, an analysis circuit 3, a drive mechanism 4, an input interface 5, an output interface 6, a communication interface 7, a storage circuit 8, and a control circuit 9.

  The automatic analyzer 1 is a device that measures the concentration of a sample or the like using a latex agglutination method, and various carrier particles can be used as an insoluble carrier to be added to a reagent. As the carrier particles, for example, latex particles, polystyrene, polystyrene latex, silica particles and the like can be used.

  The analysis mechanism 2 adds, to a standard sample or a sample such as a test sample, a reagent used in each inspection item set for the sample. The analysis mechanism 2 measures a reaction solution obtained by adding a reagent to a sample, and generates, for example, standard data represented by absorbance or light intensity and test data. In the present embodiment, the standard data represents the measurement data of the absorbance or the amount of scattered light for a standard sample whose concentration to be detected is known. Further, the test data represents measurement data of the absorbance or the amount of scattered light of the test sample.

  The analysis circuit 3 is a processor that analyzes standard data generated by the analysis mechanism 2 and test data, and generates calibration data, analysis data, and the like. The calibration data indicates, for example, the relationship between standard data and a standard calibration curve preset for a standard sample. The standard calibration curve is, for example, a calibration curve with high measurement accuracy calculated by a reagent manufacturer using a standard sample. Further, the analysis data is, for example, data represented as a concentration value and an activity value of an enzyme obtained by analyzing the test data based on the calibration data.

  The analysis circuit 3 executes an operation program stored in the storage circuit 8 and realizes functions corresponding to the operation program to generate calibration data, analysis data, and the like. For example, the analysis circuit 3 obtains standard data obtained for a standard sample whose absorbance or light intensity is known and has a concentration of 0, and a plurality of standard samples whose absorbance or light intensity is known and whose concentrations are known, respectively, Calibration data is calculated based on a standard calibration curve set in advance for a standard sample, and a photometric timing set in advance. Further, the analysis circuit 3 generates analysis data based on the test data, calibration data of an inspection item corresponding to the test data, and photometric timing set in advance. The analysis circuit 3 outputs the generated calibration data, analysis data and the like to the control circuit 9.

  The drive mechanism 4 drives the analysis mechanism 2 under the control of the control circuit 9. The drive mechanism 4 is realized by, for example, a gear, a stepping motor, a belt conveyor, and a lead screw.

  The input interface 5 receives, for example, the setting of analysis parameters and the like of each inspection item related to the sample requested to be measured from the operator or via the in-hospital network NW. The input interface 5 is realized by, for example, a mouse, a keyboard, and a touch pad to which an instruction is input by touching the operation surface. The input interface 5 is connected to the control circuit 9, converts an operation instruction input from the operator into an electric signal, and outputs the electric signal to the control circuit 9. In the present specification, the input interface 5 is not limited to one having physical operation parts such as a mouse and a keyboard. For example, an electrical signal processing circuit that receives an electrical signal corresponding to an operation instruction input from an external input device provided separately from the automatic analyzer 1 and outputs the electrical signal to the control circuit 9 is also an input interface Included in example 5

  The output interface 6 is connected to the control circuit 9 and outputs a signal supplied from the control circuit 9. The output interface 6 is realized by, for example, a display circuit, a print circuit, and the like. The display circuit includes, for example, a CRT display, a liquid crystal display, an organic EL display, an LED display, a plasma display, and the like. Note that a display circuit also includes a processing circuit that converts data representing a display target into a video signal and outputs the video signal to the outside. The printed circuit includes, for example, a printer. An output circuit that outputs data representing a print target to the outside is also included in the print circuit.

  The communication interface 7 is connected to, for example, the in-hospital network NW. The communication interface 7 performs data communication with an HIS (Hospital Information System) via the in-hospital network NW. The communication interface 7 may perform data communication with the HIS via a laboratory information system (LIS) connected to the in-hospital network NW.

  The storage circuit 8 includes a processor-readable storage medium, such as a magnetic or optical storage medium, or a semiconductor memory. The memory circuit 8 does not necessarily have to be realized by a single memory device. For example, the storage circuit 8 may be realized by a plurality of storage devices.

  The memory circuit 8 stores an operation program executed by the analysis circuit 3 and an operation program executed by the control circuit 9. The storage circuit 8 stores calibration curve information on the reagent held in the analysis mechanism 2. The calibration curve information includes data on a standard calibration curve set in advance for the reagent for each test item. Further, in the calibration curve information, for example, data on a photometric timing line preset for the reagent is included for each inspection item. Calibration curve information is provided from the reagent manufacturer via the communication interface 7, for example, in units of lots of reagents. The calibration curve information may be provided, for example, by the reagent manufacturer together with the reagent, and may be input by the operator from the input interface 5.

  The photometric timing line is a reference line that defines measurement timing. The photometric timing represents a point in time of acquiring information such as absorbance used when generating calibration data. That is, the photometric timing represents, for example, an elapsed time after adding a reagent including carrier particles, which is an insoluble carrier on which a component that binds to a detection target is immobilized, to a standard sample. In addition, the photometric timing represents a point in time of acquiring information such as absorbance used when generating analysis data. That is, the photometric timing represents, for example, an elapsed time since the reagent containing the carrier particles is added to the test sample.

  FIG. 2 is a diagram showing an example of a photometric timing line and photometric timing for a predetermined reagent. In FIG. 2, a thick solid line indicates a photometric timing line. The thin solid line represents a reaction curve obtained based on the absorbance measured after adding the reagent to a standard sample of a predetermined concentration. Of the reaction curves shown in FIG. 2, the reaction curve having a steeper rise in absorbance represents the reaction curve for a standard sample of higher concentration. Circles are points of intersection of the photometric timing line and the response curve, and represent photometric timing. In FIG. 2, the photometric timing line obliquely extends from the lower right to the upper left in the drawing. That is, the photometric timing line is set so that the photometric timing is delayed when the reagent is added to the standard sample with low concentration, and the photometric timing is advanced when the reagent is added to the standard sample with high concentration. . It should be noted that the calibration curve information may include not data related to the light measurement timing line but data related to the light measurement timing.

  The storage circuit 8 stores calibration data generated by the analysis circuit 3 for each inspection item. The storage circuit 8 stores the analysis data generated by the analysis circuit 3 for each test sample.

  The control circuit 9 is a processor that functions as the center of the automatic analyzer 1. The control circuit 9 executes an operation program stored in the storage circuit 8 to realize a function corresponding to the operation program. The control circuit 9 may have a storage area for storing at least a part of the data stored in the storage circuit 8.

  FIG. 3 is a schematic view showing an example of the configuration of the analysis mechanism 2 shown in FIG. The analysis mechanism 2 shown in FIG. 3 includes a reaction disk 201, a constant temperature unit 202, a sample disk 203, a first reagent storage 204, and a second reagent storage 205.

  The reaction disk 201 transports the reaction container 2011 along a predetermined path. Specifically, the reaction disc 201 holds a plurality of reaction vessels 2011 arranged in a ring. The reaction disc 201 is alternately rotated and stopped at predetermined time intervals by the drive mechanism 4.

  The reaction container 2011 is formed of, for example, glass. The reaction vessel 2011 has a square pole shape and an opening at the top. The light emitted from the light source provided to the photometric unit 214 is incident from the outer surface of the first side wall among the first to fourth side walls forming the quadrangular prism. The light incident from the outer surface of the first side wall is emitted from the outer surface of the second side wall facing the first side wall among the first to fourth side walls.

  The constant temperature unit 202 stores the heat medium set to a predetermined temperature. The constant temperature unit 202 causes the reaction container 2011 to be immersed in a heat medium to be stored, thereby raising the temperature of the reaction liquid contained in the reaction container 2011.

  The sample disk 203 holds a plurality of sample containers for storing a sample. The sample disc 203 is rotated by the drive mechanism 4.

  The first reagent storage 204 cools a plurality of reagent containers containing a standard sample and a first reagent that reacts with a predetermined component contained in the test sample. The first reagent is, for example, a buffer solution containing bovine serum albumin (BSA) or the like. A reagent label is attached to the reagent container. The reagent label is printed with an optical mark indicating reagent information. For the optical mark, for example, an arbitrary pixel code such as a one-dimensional pixel code and a two-dimensional pixel code is used. The reagent information is information related to the reagent contained in the reagent container, and includes, for example, a reagent name, a reagent maker code, a reagent item code, a bottle type, a bottle size, a capacity, a production lot number, and a term of validity.

  Further, the first reagent storage 204 cools a plurality of standard sample containers for storing standard samples. The standard sample container may contain standard samples of the same component having different concentrations. The standard sample container may be held by the sample disc 203.

  In the first reagent storage 204, a reagent rack 2041 is rotatably provided. The reagent rack 2041 holds a plurality of reagent containers and a plurality of standard sample containers arranged in an annular shape. The reagent rack 2041 is rotated by the drive mechanism 4. In addition, in the first reagent storage 204, a reader (not shown) for reading reagent information from a reagent label attached to a reagent container is provided. The read reagent information is stored in the storage circuit 8.

  A first reagent suction position is set at a predetermined position on the first reagent storage 204. The first reagent aspirating position is, for example, at a position where the rotation trajectory of the first reagent dispensing probe 209 intersects with the movement trajectory of the opening of the reagent container and the standard sample container arranged annularly in the reagent rack 2041. Provided.

  The second reagent storage 205 cools a plurality of reagent containers storing the second reagent that makes a pair with the first reagent of the two reagent system. The second reagent is a solution containing a predetermined antigen or antibody contained in the sample and an insoluble carrier, eg, carrier particles, on which the antigen or antibody that binds or separates due to a specific antigen-antibody reaction is immobilized. It may be an enzyme, a substrate, an aptamer, or a receptor as one that binds or separates by a specific reaction. In the second reagent storage 205, a reagent rack 2051 is rotatably provided.

  The reagent rack 2051 holds a plurality of reagent containers arranged in an annular shape. In the second reagent storage 205, the standard sample container for storing the standard sample may be cooled. The reagent rack 2051 is rotated by the drive mechanism 4. In the second reagent storage 205, a reader (not shown) for reading reagent information from a reagent label attached to the reagent container is provided. The read reagent information is stored in the storage circuit 8.

  A second reagent suction position is set at a predetermined position on the second reagent storage 205. The second reagent suction position is provided, for example, at a position where the rotation track of the second reagent dispensing probe 211 intersects with the movement track of the opening of the reagent container arranged annularly in the reagent rack 2051.

  Further, the analysis mechanism 2 shown in FIG. 3 includes a sample dispensing arm 206, a sample dispensing probe 207, a first reagent dispensing arm 208, a first reagent dispensing probe 209, a second reagent dispensing arm 210, a second A reagent dispensing probe 211, a first stirring unit 212, a second stirring unit 213, a photometric unit 214, and a washing unit 215 are provided.

  The sample dispensing arm 206 is provided between the reaction disc 201 and the sample disc 203. The sample dispensing arm 206 is vertically vertically movable and horizontally rotatable by the drive mechanism 4. The sample dispensing arm 206 holds the sample dispensing probe 207 at one end.

  The sample dispensing probe 207 pivots along an arc-shaped pivoting track as the sample dispensing arm 206 pivots. The opening portion of the sample container held by the sample disc 203 is positioned on this rotating orbit. A sample discharge position for discharging the sample sucked by the sample dispensing probe 207 to the reaction container 2011 is provided on the rotation trajectory of the sample dispensing probe 207. The sample discharge position corresponds to the intersection of the rotation trajectory of the sample dispensing probe 207 and the movement trajectory of the reaction container 2011 held by the reaction disk 201.

  The sample dispensing probe 207 is driven by the drive mechanism 4 and moves vertically on the opening of the sample container held by the sample disk 203 or at the sample discharge position. Further, the sample dispensing probe 207 sucks the sample from the sample container located immediately below according to the control of the control circuit 9. Further, the sample dispensing probe 207 discharges the aspirated sample to the reaction container 2011 located immediately below the sample discharge position according to the control of the control circuit 9.

  The first reagent dispensing arm 208 is provided near the outer periphery of the first reagent storage 204. The first reagent dispensing arm 208 is vertically movable in the vertical direction by the drive mechanism 4 and rotatably provided in the horizontal direction. The first reagent dispensing arm 208 holds the first reagent dispensing probe 209 at one end.

  The first reagent dispensing probe 209 pivots along an arc-shaped pivoting track as the first reagent dispensing arm 208 pivots. A first reagent suction position is provided on the rotation track. In addition, the first reagent discharge position for discharging the first reagent sucked by the first reagent dispensing probe 209 or the standard sample to the reaction container 2011 is set on the rotation trajectory of the first reagent dispensing probe 209 It is done. The first reagent discharge position corresponds to an intersection point of the rotation trajectory of the first reagent dispensing probe 209 and the movement trajectory of the reaction container 2011 held by the reaction disk 201.

  The first reagent dispensing probe 209 is driven by the drive mechanism 4 and moves up and down at the first reagent suction position or the first reagent discharge position on the rotation track. Further, the first reagent dispensing probe 209 aspirates the first reagent or the standard sample from the reagent container positioned immediately below the first reagent aspirating position under the control of the control circuit 9. In addition, the first reagent dispensing probe 209 discharges the aspirated first reagent or standard sample to the reaction container 2011 located immediately below the first reagent discharge position according to the control of the control circuit 9. That is, the first reagent dispensing probe 209 is an example of the discharge unit in the present embodiment.

  The second reagent dispensing arm 210 is provided near the outer periphery of the first reagent storage 204. The second reagent dispensing arm 210 is vertically movable in the vertical direction by the drive mechanism 4 and rotatably provided in the horizontal direction. The second reagent dispensing arm 210 holds the second reagent dispensing probe 211 at one end.

  The second reagent dispensing probe 211 pivots along an arc-shaped pivoting track as the second reagent dispensing arm 210 pivots. A second reagent suction position is provided on the rotation track. In addition, on the rotation trajectory of the second reagent dispensing probe 211, a second reagent discharge position for discharging the second reagent sucked by the second reagent dispensing probe 211 to the reaction container 2011 is set. The second reagent discharge position corresponds to an intersection point of the rotation trajectory of the second reagent dispensing probe 211 and the movement trajectory of the reaction container 2011 held by the reaction disk 201.

  The second reagent dispensing probe 211 is driven by the drive mechanism 4 and vertically moves at a second reagent suction position or a second reagent discharge position on the rotation track. Further, the second reagent dispensing probe 211 aspirates the second reagent from the reagent container located immediately below the second reagent aspirating position under the control of the control circuit 9. In addition, the second reagent dispensing probe 211 discharges the aspirated second reagent to the reaction container 2011 positioned immediately below the second reagent discharge position according to the control of the control circuit 9. That is, the second reagent dispensing probe 211 is an example of the discharge unit in the present embodiment.

  The first stirring unit 212 is provided near the outer periphery of the reaction disc 201. The first stirring unit 212 has a first stirring arm 2121 and a first stirring bar provided at the tip of the first stirring arm 2121. The first stirring unit 212 stirs the standard sample and the first reagent contained in the reaction container 2011 located at the first stirring position on the reaction disc 201 by the first stirring element. Further, the first stirring unit 212 stirs the sample and the first reagent contained in the reaction container 2011 positioned at the first stirring position on the reaction disc 201 by the first stirring element.

  The second stirring unit 213 is provided in the vicinity of the outer periphery of the reaction disk 201. The second stirring unit 213 has a second stirring arm 2131 and a second stirring bar provided at the tip of the second stirring arm 2131. The second stirring unit 213 stirs the standard sample, the first reagent, and the second reagent contained in the reaction container 2011 positioned at the second stirring position on the reaction disc 201 by the second stirring element. In addition, the second stirring unit 213 stirs the sample, the first reagent, and the second reagent stored in the reaction container 2011 positioned at the second stirring position by the second stirring element.

  The photometry unit 214 optically measures the reaction solution of the sample, the first reagent, and the second reagent discharged into the reaction container 2011. The photometry unit 214 includes a light source and a light detector. The photometry unit 214 emits light from the light source according to the control of the control circuit 9. The irradiated light is incident from the first side wall of the reaction vessel 2011 and emitted from the second side wall opposite to the first side wall. The photometric unit 214 detects the light emitted from the reaction container 2011 by the light detector.

  Specifically, for example, the light detector is disposed at a position on the optical axis of the light emitted from the light source to the reaction container 2011. The photodetector detects light transmitted through the reaction solution of the standard sample, the first reagent, and the second reagent in the reaction container 2011, and generates standard data represented by absorbance based on the detected light intensity. . In addition, the light detector detects the light transmitted through the reaction solution of the test sample, the first reagent, and the second reagent in the reaction container 2011, and the test represented by the absorbance based on the detected light intensity. Generate data. The photometry unit 214 outputs the generated standard data and test data to the analysis circuit 3.

  The cleaning unit 215 cleans the inside of the reaction container 2011 in which the measurement of the reaction liquid has been completed by the photometry unit 214.

  The control circuit 9 shown in FIG. 1 implements the function corresponding to the program by executing the operation program stored in the memory circuit 8. For example, the control circuit 9 has a system control function 91, a calibration control function 92, and a measurement control function 93 by executing an operation program. Although the case where the system control function 91, the calibration control function 92, and the measurement control function 93 are realized by a single processor is described in this embodiment, the present invention is not limited thereto. For example, a plurality of independent processors may be combined to constitute a control circuit, and each processor may execute an operation program to implement the system control function 91, the calibration control function 92, and the measurement control function 93.

  The system control function 91 is a function that controls each part in the automatic analyzer 1 in an integrated manner based on input information input from the input interface 5.

  The calibration control function 92 is a function to control the analysis mechanism 2 and the drive mechanism 4 so as to generate standard data. Specifically, the control circuit 9 executes the calibration control function 92 at a predetermined timing. The predetermined timing is, for example, at the time of initial setting, at the time of apparatus startup, at the time of maintenance, or when an instruction to start the calibration operation is input from the operator.

  When the calibration control function 92 is executed, the control circuit 9 controls the analysis mechanism 2 and the drive mechanism 4. By controlling the analysis mechanism 2 and the drive mechanism 4, the analysis mechanism 2 generates standard data. Specifically, for example, by driving by the driving mechanism 4, the first reagent dispensing probe 209 of the analysis mechanism 2 aspirates the standard sample from the first reagent storage 204, and the aspirated standard sample is the reaction container 2011. To discharge. The first reagent dispensing probe 209 sucks the first reagent from the first reagent storage 204, and discharges the sucked first reagent to the reaction container 2011 from which the standard sample is discharged. The first stirring unit 212 stirs the solution in which the first reagent is added to the standard sample.

  The second reagent dispensing probe 211 aspirates the second reagent from the second reagent reservoir 205, and discharges the aspirated second reagent to a mixed solution in which the standard sample and the first reagent are mixed. The second stirring unit 213 stirs the solution in which the second reagent is added to the mixed solution. The photometry unit 214 generates standard data by optically measuring the reaction solution in which the standard sample, the first reagent, and the second reagent are agitated. The photometry unit 214 outputs the generated standard data to the analysis circuit 3. The photometry unit 214 repeats measurement of the reaction solution a preset number of times in a preset cycle, and outputs the generated standard data to the analysis circuit 3. The analysis mechanism 2 repeats the above operation with reference samples of a plurality of concentrations set in advance, and outputs the generated standard data to the analysis circuit 3.

  The measurement control function 93 is a function that controls the analysis mechanism 2 and the drive mechanism 4 so as to generate test data. Specifically, the control circuit 9 executes the measurement control function 93 in accordance with a predetermined instruction. The predetermined instruction is, for example, an instruction to start measurement operation input from the operator, an instruction indicating that a predetermined time has been reached, or the like.

  When the measurement control function 93 is executed, the control circuit 9 controls the analysis mechanism 2 and the drive mechanism 4. By controlling the analysis mechanism 2 and the drive mechanism 4, test data is generated in the analysis mechanism 2. Specifically, by being driven by the drive mechanism 4, the sample dispensing probe 207 of the analysis mechanism 2 sucks the test sample from the sample disk 203 and discharges the suctioned test sample to the reaction container 2011. The first reagent dispensing probe 209 aspirates the first reagent from the first reagent storage 204, and discharges the aspirated first reagent to the reaction container 2011 from which the test sample is discharged. The first stirring unit 212 stirs the solution in which the first reagent is added to the test sample.

  The second reagent dispensing probe 211 aspirates the second reagent from the second reagent reservoir 205, and discharges the aspirated second reagent to a mixed solution in which the test sample and the first reagent are mixed. The second stirring unit 213 stirs the solution in which the second reagent is added to the mixed solution. The photometry unit 214 generates test data by optically measuring the reaction solution in which the test sample, the first reagent, and the second reagent are agitated. The photometry unit 214 outputs the generated test data to the analysis circuit 3. The photometry unit 214 repeats the measurement of the reaction solution a preset number of times in a preset cycle, and outputs the generated test data to the analysis circuit 3.

  The analysis circuit 3 shown in FIG. 1 implements the function corresponding to the program by executing the operation program stored in the memory circuit 8. For example, the analysis circuit 3 has a calibration data generation function 31 and an analysis data generation function 32 by executing an operation program. In the present embodiment, although the case where the calibration data generation function 31 and the analysis data generation function 32 are realized by a single processor will be described, the present invention is not limited thereto. For example, the calibration data generation function 31 and the analysis data generation function 32 may be realized by configuring an analysis circuit by combining a plurality of independent processors and executing the operation program by each processor.

  The calibration data generation function 31 is a function of generating calibration data based on the standard data generated by the analysis mechanism 2. Specifically, when the analysis circuit 3 receives the standard data generated by the analysis mechanism 2, the analysis circuit 3 executes the calibration data generation function 31. When the calibration data generation function 31 is executed, the analysis circuit 3 reads from the storage circuit 8 data on a standard calibration curve set in advance for the reagent of a predetermined inspection item and data on a photometric timing line. The analysis circuit 3 generates calibration data based on the standard data, the standard calibration curve, and the photometric timing line. The analysis circuit 3 stores the generated calibration data in the storage circuit 8.

  The analysis data generation function 32 is a function of generating analysis data by analyzing the test data generated by the analysis mechanism 2. Specifically, when the analysis circuit 3 receives the test data generated by the analysis mechanism 2, the analysis circuit 3 executes an analysis data generation function 32. When the analysis data generation function 32 is executed, the analysis circuit 3 reads out from the storage circuit 8 calibration data stored for a predetermined inspection item and data on a photometric timing line. The analysis circuit 3 generates analysis data based on the test data, the photometric timing line, and the calibration data.

(Setting of standard calibration curve, photometric timing line, and photometric timing)
Next, an example of setting of the standard calibration curve, the photometric timing line, and the photometric timing will be described in detail. In the following description, for example, a case where the standard calibration curve, the photometric timing line, and the photometric timing are set based on the measured absorbance is measured by using the above-described automatic analyzer 1 in a reagent manufacturer, for example. Do.

  First, the photometric timing and the setting of the photometric timing line will be described.

  When designing or prescribing reagents, a time course for a standard sample is obtained. For example, standard samples S1 to S8 having known concentrations are respectively dispensed into the reaction containers 2011-1 to 2011-8. For example, the concentration of standard sample S1 is 0, the concentration of standard sample S2 is 8 ng / ml, the concentration of standard sample S3 is 32 ng / ml, the concentration of standard sample S4 is 64 ng / ml, and the standard sample is The concentration of S5 is 129 ng / ml, the concentration of standard sample S6 is 259 ng / ml, the concentration of standard sample S7 is 518 ng / ml, and the concentration of standard sample S8 is 1037 ng / ml.

  Subsequently, the first reagent which is a buffer solution is added to the standard samples S1 to S8 respectively dispensed to the reaction containers 2011-1 to 2011-8. The standard samples S1 to S8 to which the first reagent is added are agitated. After stirring, the mixed solution contained in the reaction containers 2011-1 to 2011-8 is incubated at a predetermined temperature for a predetermined period. After the incubation, a second reagent containing carrier particles on which a component to be bound to the detection target in the standard sample is immobilized is added to the liquid mixtures respectively accommodated in the reaction containers 2011-1 to 2011-8. The mixture to which the second reagent is added is stirred.

  The reaction solutions of the standard samples S1 to S8, the first reagent, and the second reagent stored in the reaction vessels 2011-1 to 2011-8 are irradiated with light while being incubated at a predetermined temperature for a predetermined period. . Specifically, for example, in the reaction containers 2011-1 to 2011-8, after the second reagent is added to the standard samples S1 to S8, light is emitted from the light source 33 times in a cycle of 4.6 seconds. Be done. The transmitted light transmitted through the reaction containers 2011-1 to 2011-8 is detected by the photodetector. Based on the detected light intensity, a time course for the standard sample is obtained.

  FIG. 4 shows an example of measured values and time courses (response curves) for standard samples S1 to S8. In FIG. 4, the absorbance measured in the reaction solution for standard samples S1 to S8 having different concentrations is shown in 17 cycles to 33 cycles. In FIG. 4, square marks indicate measured values of absorbance, and lines connecting the square marks indicate reaction curves. The reaction curve C1 is calculated based on the absorbance of each cycle of the standard sample S1. Similar to the reaction curve C1, the reaction curves C2 to C8 are calculated based on the absorbance of each cycle of the standard samples S2 to S8. For the standard samples S2 to S4 with low concentrations, the increase in reaction time and the increase in absorbance are in a substantially proportional relationship. On the other hand, for the standard samples S5 to S8 with medium to high concentrations, the absorbance approaches saturation when the reaction time passes.

  The agglutination reaction of the carrier particles contained in the second reagent has a characteristic that the absorbance is saturated when the concentration of the sample increases and the hook phenomenon occurs. Therefore, it is desirable that the reaction solution with a standard sample having a high concentration be measured at a photometric timing shorter than that causing the hook phenomenon. In the present embodiment, based on the response curves C1 to C8, for example, the photometric timing line is set such that the photometric timing becomes shorter as the density of the standard sample changes from low density to high density. When there is a portion where the response curves intersect, the photometric timing is set earlier than the time when the response curves intersect.

  More specifically, for example, first, a measurement value extracted one by one from each of the reaction curves C1 to C8 is used as a data set, and a regression analysis using absorbance as a target variable is performed. The regression analysis is repeated while changing the measurement value to be extracted, and a regression equation satisfying a predetermined condition is extracted among the calculated plurality of regression equations. The predetermined condition includes, for example, that the determination coefficient of the regression equation is 0.9 or more, and that the order of the regression equation is less than or equal to a predetermined order. Then, the intersection with the reaction curves C1 to C8 is determined for each extracted regression equation, and the sum, the product or the like of the difference in absorbance between the intersections with the adjacent reaction curves is calculated. Then, among the extracted regression equations, the regression equation in which the value of the calculated sum or product is the largest is taken as the photometric timing line.

  FIGS. 5 and 6 are diagrams showing examples of photometric timing lines set based on the response curves C1 to C8 shown in FIG. 4 and photometric timing. In FIG. 5, the photometric timing line is represented by a thick broken line and has a linear shape. In FIG. 6, the photometric timing line is represented by a thick broken line and has a curved shape. In FIG. 5 and FIG. 6, circle marks indicate photometric timing. The photometric timing is set at the intersection of the photometric timing line and the response curves C1 to C8.

  The photometric timing line is not limited to a line based on regression as shown in FIGS. 5 and 6. The photometric timing line may be set based on the measurement value used for regression analysis. Specifically, for example, a regression equation satisfying a predetermined condition is extracted from among a plurality of regression equations calculated by regression analysis. Measured values for each of the reaction curves C1 to C8 used in the regression analysis are acquired for each extracted regression equation, and the sum or product or the like of the difference in absorbance between the measured values of adjacent reaction curves is calculated. Then, a line connecting the calculated sum or the measured value with the largest product value is taken as a photometric timing line.

  FIG. 7 is a diagram illustrating an example of a photometric timing line set based on measurement values used for regression analysis, and photometric timing. As described above, it becomes possible to set the photometric timing line using the actually measured values.

  Next, generation of a standard calibration curve will be described. The standard calibration curve is generated based on, for example, the absorbance measured at the photometric timing set for each of the reaction curves C1 to C8. The standard calibration curve represents the correlation between concentration and absorbance.

  FIG. 8 is a diagram showing an example of a standard calibration curve generated based on the photometric timing shown in FIG. According to the standard calibration curve shown in FIG. 8, the absorbance increases with the increase of the concentration to be detected. Therefore, regardless of the level of the concentration to be detected, the concentration to be detected can be uniquely determined from the absorbance.

  The photometry timing line set in the present embodiment is not limited to one photometry timing line as shown in FIGS. 5 to 7. A plurality of photometric timing lines may be set according to the density of the standard sample. For example, as shown in FIG. 9, two photometric timing lines L1 and L2 may be set. At this time, the photometric timings T1 to T3 for the low density standard samples S1 to S3 are set at the intersections of the response curves C1 to C3 for the standard samples S1 to S3 and the photometric timing line L1. Further, the photometric timings T4 to T8 for the medium to high concentration standard samples S4 to S8 are set at the intersections of the reaction curves C4 to C8 for the standard samples S4 to S8 and the photometric timing line L2. In FIG. 9, the frequencies of light to be irradiated may be changed between the low concentration band and the high concentration band.

  FIG. 10 is a diagram showing an example of a standard calibration curve generated based on the photometric timing shown in FIG. The standard calibration curve shown in FIG. 10 represents the correlation between concentration and absorbance. The absorbance shown in FIG. 10 is calculated, for example, from the difference between the absorbance in the initial cycle in which the reaction in the reaction solution has not progressed and the absorbance measured at photometric timings T1 to T8 shown in FIG. According to FIG. 10, for each concentration band to be detected, a standard calibration curve is generated in which the absorbance increases with the increase of the concentration to be detected. This makes it possible to refer to the standard calibration curve for the middle and high concentration band for measurement in the short cycle, and to refer to the standard calibration curve for the low concentration band for the measurement in the long cycle. Can be determined uniquely.

  In the present embodiment, the photometric timing line and the photometric timing are set as an example based on the measurement results using the standard samples S1 to S8. However, the present invention is not limited to this. The number of standard samples to be used may be eight or less, for example, about six.

  As described above, in the first embodiment, to each of the standard samples S1 to S8, a reagent including an insoluble carrier on which a component that binds to the detection target in the standard samples S1 to S8 is immobilized is added. Each reaction solution in which the reagent is added to the standard samples S1 to S8 is optically measured at different photometric timings. Then, a calibration curve as a standard calibration curve is generated based on the photometric result of each standard sample. As a result, it is possible to generate a calibration curve using photometric results acquired at different photometric timings for each concentration.

  In the first embodiment, the photometric timing is set so as to be gradually delayed as the density of the standard sample changes from the high density to the low density. As a result, a calibration curve is generated using photometric results acquired earlier as the concentration is higher.

  Further, in the first embodiment, an intensity curve representing a temporal change in photometric intensity is calculated for each standard sample. Based on the intensity curve for each standard sample, the photometric timing line is set such that the time point for acquiring information is gradually delayed from the high concentration standard sample to the low concentration standard sample. The intersection point of the intensity curve of each standard sample and the photometric timing line is used as the photometric timing. This makes it possible to obtain information for generating a calibration curve in a reaction time suitable for each concentration to be detected.

(Generation of calibration data by automatic analyzer 1)
Next, an example of an operation in which the automatic analyzer 1 configured as described above generates calibration data will be described in detail.

  First, the automatic analyzer 1 acquires calibration curve information of the second reagent received in lot units, for example, via a network. The calibration curve information includes data on a standard calibration curve of the second reagent received in lot units and data on a photometric timing line. The acquired calibration curve information is stored in the storage circuit 8.

  The control circuit 9 executes, for example, the calibration control function 92 when the automatic analyzer 1 is started. When the calibration control function 92 is executed, the control circuit 9 controls the analysis mechanism 2 and the drive mechanism 4 to generate standard data for a predetermined inspection item. Specifically, for example, the opening of the container of at least one standard sample of standard samples S1 to S8 held in the reagent rack 2041 by rotating the reagent rack 2041 provided in the first reagent storage 204 Is placed at the first reagent aspiration position. The standard sample stored in the container is aspirated by the first reagent dispensing probe 209. The aspirated standard sample is discharged to the reaction container 2011.

  Subsequently, the reagent rack 2041 is rotated, and the opening portion of the reagent container of the first reagent held by the reagent rack 2041 is disposed at the first reagent suction position. The first reagent stored in the reagent container is aspirated by the first reagent dispensing probe 209. The aspirated first reagent is discharged to the reaction container 2011 from which the standard sample is discharged. The reaction container 2011 from which the standard sample and the first reagent have been discharged is transported by the reaction disk 201 to the first stirring position. The solution in which the first reagent is added to the standard sample is stirred by the first stirring unit 212 and incubated at a predetermined temperature for a predetermined period.

  Subsequently, the reagent rack 2051 provided in the second reagent storage 205 is rotated, and the opening portion of the reagent container of the second reagent held in the reagent rack 2051 is disposed at the second reagent suction position. The second reagent stored in the reagent container is aspirated by the second reagent dispensing probe 211. The aspirated second reagent is discharged into a mixed solution in which the standard sample and the first reagent are mixed. The reaction container 2011 from which the standard sample, the first reagent, and the second reagent are discharged is transported by the reaction disc 201 to the second stirring position. The solution in which the first reagent and the second reagent are added to the standard sample is stirred by the second stirring unit 213.

  The reaction container 2011 stirred by the second stirring unit 213 is transported by the reaction disk 201 to the photometric point. The light source of the photometry unit 214 applies light to the reaction container 2011 transported to the photometric point. The light detector of the photometry unit 214 detects the light transmitted through the reaction container 2011. Based on the detected light intensity, standard data as absorbance are generated. The generated standard data is output to the analysis circuit 3. The measurement by the photometry unit 214 is repeated a preset number of times in a preset cycle, and the generated standard data is output to the analysis circuit 3. The standard sample used to generate standard data is not limited to one. For example, two standard samples may be used. At this time, for example, standard samples S2 and S5 are used.

  When the analysis circuit 3 receives the standard data output from the analysis mechanism 2, the analysis circuit 3 executes the calibration data generation function 31. When the calibration data generation function 31 is executed, the analysis circuit 3 reads from the storage circuit 8 data on a standard calibration curve for a predetermined test item of the second reagent and data on a photometric timing line. The analysis circuit 3 calculates a reaction curve by drawing an approximate curve with respect to the measurement value of the standard sample included in the standard data. The analysis circuit 3 calculates the absorbance at the light measurement timing (the light measurement timings T2 and T5 when using the standard samples S2 and S5) which is the intersection of the reaction curve and the light measurement timing line.

  The analysis circuit 3 corrects and interpolates the standard calibration curve based on the absorbance at the photometric timing to obtain a calibration curve that enables measurement of the test data of the test item. The analysis circuit 3 stores, for example, the corrected calibration curve in the storage circuit 8 as calibration data. The method of obtaining a calibration curve by correcting a standard calibration curve based on standard data obtained by measurement of at least one standard sample is a commonly used method from the viewpoint of saving reagents. However, it is not limited to this. The analysis circuit 3 may acquire a calibration curve without using a standard calibration curve.

  Specifically, for example, measurements on all the standard samples S1 to S8 are performed to generate standard data as absorbance. When the analysis circuit 3 receives the standard data output from the analysis mechanism 2, the analysis circuit 3 executes the calibration data generation function 31. When the calibration data generation function 31 is executed, the analysis circuit 3 reads out from the storage circuit 8 data on a photometric timing line for a predetermined inspection item of the second reagent. The analysis circuit 3 calculates a reaction curve by drawing an approximate curve with respect to the measurement value of the standard sample included in the standard data. The analysis circuit 3 calculates the absorbance at the light measurement timing which is an intersection point of the reaction curve and the light measurement timing line. The analysis circuit 3 obtains a calibration curve based on the absorbance at the photometric timing. The analysis circuit 3 stores, for example, a calibration curve in the storage circuit 8 as calibration data. This makes it possible to obtain a more appropriate calibration curve under the current environment.

  As described above, in the first embodiment, the automatic analyzer 1 causes the first reagent dispensing probe 209 to cause at least one of the standard samples S1 to S8 a component to be bound to the detection target contained in the standard sample. Add a reagent containing the immobilized insoluble carrier. The photometry unit 214 obtains the photometric intensity of the standard sample by optically measuring the reaction solution in which the reagent is added to the standard sample. The memory circuit 8 stores a photometric timing line which is set such that the point of time when information on photometric intensity is acquired becomes gradually longer from a high density standard sample to a low density standard sample. Then, the analysis circuit 3 acquires an intensity curve representing the temporal change of the photometric intensity based on the acquired photometric intensity by the calibration data generation function 31, and uses information acquired at the intersection of the intensity curve and the photometric timing line. Calibration data is generated on the basis of this. Thereby, the automatic analyzer 1 can generate calibration data with a wide dynamic range by acquiring information at a reaction time suitable for the concentration of the detection target.

(Generation of analytical data by automatic analyzer 1)
Next, an example of an operation in which the automatic analyzer 1 generates analysis data will be described in detail.

  The control circuit 9 executes the measurement control function 93, for example, when an instruction to start the measurement operation is input from the operator. When the measurement control function 93 is executed, the control circuit 9 controls the analysis mechanism 2 and the drive mechanism 4 to generate test data for a predetermined inspection item.

  When the analysis circuit 3 receives the test data output from the analysis mechanism 2, the analysis circuit 3 executes an analysis data generation function 32. When the analysis data generation function 32 is executed, the analysis circuit 3 reads out from the storage circuit 8 calibration data stored for the inspection item corresponding to the test data and data on the photometric timing line. The analysis circuit 3 calculates a test data approximate curve by drawing an approximate curve with respect to a plurality of actually measured absorbances included in the test data. The test data approximate curve is not limited to the approximate curve calculated based on the absorbance, and may be a straight line obtained by linearly interpolating between the absorbances. It is premised that the accuracy of the approximation equation is high, but the one represented by the approximation curve is more robust to noise than the one represented by the straight line.

  FIG. 11 is a diagram showing examples of reaction curves C1 to C8, standard photometric timing lines, and test data approximate curves calculated based on test data for standard samples S1 to S8. In the example shown in FIG. 11, the test data approximate curve is represented by a thick two-dot chain line, and intersects the photometric timing line between photometric timings T4 and T5.

  In order to determine the concentration of the detection target in the sample based on the test data, it is necessary to obtain the position of the intersection of the photometric timing line and the test data approximate curve, and replace the position on the calibration data. In the analysis circuit 3, for example, the position on the calibration data is specified by the following processing.

  FIG. 12 is a diagram illustrating an example of the corrected calibration data stored in the memory circuit 8. First, the analysis circuit 3 connects T4 and T5 on the photometric timing line shown in FIG. 11 with a straight line. When a point at which the photometric timing line intersects the test data approximate curve is X, it is assumed that T4X: XT5 = a: b. The analysis circuit 3 calculates the point at which the line segment T4T5 is divided into a: b on the corrected calibration data of the inspection item corresponding to the test data. This point is the intersection point X in which the intersection point on the photometric timing line is replaced on the calibration data. In FIG. 12, a vertical line is drawn from the intersection point X to the x axis of the graph, and the value intersecting the x axis is the concentration of the detection target in the sample.

  As described above, in the first embodiment, the automatic analyzer 1 adds the reagent including the insoluble carrier on which the predetermined component is immobilized to the test sample by the first reagent dispensing probe 209. The photometry unit 214 optically measures the reaction solution in which the reagent is added to the test sample to obtain the photometric intensity of the test sample. Then, the analysis circuit 3 acquires the intensity curve representing the temporal change of the photometric intensity based on the acquired photometric intensity of the test sample by the analysis data generation function 32, and acquires it at the intersection of the intensity curve and the photometry timing line. The analysis data is generated based on the information to be analyzed and the calibration data. As a result, the automatic analyzer 1 can generate analytical data using calibration data with a wide dynamic range.

Second Embodiment
In the first embodiment, the standard calibration curve and the calibration curve are described as representing, for example, the correlation between concentration and absorbance. In the second embodiment, the case where the standard calibration curve and the calibration curve represent, for example, the correlation between concentration and reaction time will be described.

(Setting of standard calibration curve, photometric timing line, and photometric timing)
In the second embodiment, the standard calibration curve represents, for example, the correlation between concentration and reaction time. The standard calibration curve is generated, for example, based on the reaction time until reaching the photometric timing set for each of the reaction curves C1 to C8 in a reagent maker or the like.

  Specifically, for example, when the photometric timing line has a curved shape as shown in FIG. 6, the reaction time until reaching each of the photometric timings T1 to T8 is expressed as shown in FIG. At this time, the standard calibration curve is calculated with the obtained reaction time as the ordinate and the concentration as the abscissa. FIG. 14 is a diagram showing an example of a standard calibration curve represented from concentration and reaction time, calculated based on FIG. According to the standard calibration curve shown in FIG. 14, the reaction time decreases with the increase of the concentration to be detected.

  In the second embodiment, the photometric timing defined by the photometric timing line is not limited to being set within the maximum number of measurements. That is, the photometric timing may be set, for example, to a time later than 33 cycles.

  For example, based on at least one of the time courses (reaction curves) for the standard sample, reaction curves in sections exceeding the maximum number of measurements are estimated. Then, a photometric timing line is set based on the response curve and the estimated response curve.

  Specifically, for example, reaction curves C1 to C8 for standard samples S1 to S8 shown in FIG. 4 are assumed. For the reaction curves C1 to C8, for example, linear regression analysis using four points of observation intervals 30 to 33 is performed. The section used for linear regression analysis is arbitrary and can be selected by the user and the reagent maker.

  Among the approximate straight lines calculated by linear regression analysis, an approximate straight line having a determination coefficient of, for example, 0.5 or more is extracted. This is because when the determination coefficient of the approximate straight line calculated by the four points in the observation sections 30 to 33 is a negative value or less than 0.5, curve approximation is more desirable than linear approximation. The extracted approximate straight line represents the reaction after 33 cycles for the reaction curve corresponding to the approximate straight line. For response curves for which an approximation line with a coefficient of determination less than 0.5 was calculated, responses after 33 cycles are not considered.

  For example, measurement values extracted one by one from each of the reaction curves C1 to C8 including a section after 33 cycles are used as a data set, and regression analysis using absorbance as a target variable is performed. The regression analysis is repeated while changing the measurement value to be extracted, and a regression equation satisfying a predetermined condition is extracted among the calculated plurality of regression equations. Then, the intersection with the reaction curves C1 to C8 is determined for each extracted regression equation, and the sum, the product or the like of the difference in absorbance between the intersections with the adjacent reaction curves is calculated. Then, among the extracted regression equations, the regression equation in which the value of the calculated sum or product is the largest is taken as the photometric timing line.

  FIG. 15 is a diagram illustrating an example of photometric timing lines set based on the response curves C1 to C8 including a section after 33 cycles, and photometric timing. In FIG. 15, the photometric timing line is represented by a thick broken line and has a curved shape. In FIG. 15, circle marks indicate photometric timing. In FIG. 15, the approximate straight lines calculated for the reaction curves C1 to C4 represent the reactions after 33 cycles. In FIG. 15, the photometric timing line and the response curves C1 to C3 intersect in a section exceeding 33 cycles. That is, photometric timings T1 to T3 for the standard samples S1 to S3 occur in a section exceeding 33 cycles. A standard calibration curve is generated based on the reaction time until reaching the photometric timings T1 to T8 shown in FIG.

  FIG. 16 is a diagram showing an example of a standard calibration curve generated based on the photometric timing shown in FIG. According to the standard calibration curve shown in FIG. 16, the reaction time decreases with the increase of the concentration to be detected.

  In addition, about the reaction curve in which the approximation line less than the coefficient of determination 0.5 was calculated, an approximation curve is newly calculated using four points of observation sections 30 to 33, and the reaction after the 33 cycles is calculated by the calculated approximation curve. It does not matter. Further, the threshold of the determination coefficient when extracting the approximate straight line is not limited to 0.5. The threshold can be set arbitrarily.

  Also, the photometric timing line is not limited to being set after calculating the reaction curve after 33 cycles. For example, the photometric timing line may be set before calculating the response curve after 33 cycles. For example, it is determined for each reaction curve which point the absorbance value becomes (reaction time) as a point on the standard calibration curve. A metering timing line is set based on the determined point.

  At this time, whether or not it is necessary to calculate a reaction curve after 33 cycles can be determined based on the set photometric timing line. For example, when the intersection point between the set photometric timing line and the response curve does not exist within 33 cycles, it is determined that it is necessary to calculate the response curve after 33 cycles. When it is necessary to calculate a reaction curve after 33 cycles, regression analysis is performed using measured values in a predetermined section of the reaction curve, and a reaction curve after 33 cycles is calculated.

  In the second embodiment, the photometric timing line may not be a line extending diagonally from the lower right to the upper left. For example, the photometric timing line may be a straight line parallel to the horizontal axis.

  For example, the photometric timing line is set as a straight line with fixed absorbance. FIG. 17 is a diagram illustrating an example of a photometric timing line whose absorbance is fixed, and the photometric timing. In FIG. 17, the photometric timing line is represented by a thick broken line, and the photometric timing is represented by a circle. In FIG. 17, the approximate straight lines calculated for the reaction curves C1 to C4 represent reactions after 33 cycles. In FIG. 17, the photometric timing line and the response curves C2 and C3 cross each other in a section exceeding 33 cycles. Note that, since the reaction curve C2 has a predetermined angle with respect to the horizontal axis, when these reaction curves are stretched, they will eventually intersect the photometric timing line. As a result, photometric timings T2 and T3 for the standard samples S2 and S3 are generated in a section exceeding 33 cycles. A standard calibration curve is generated based on the reaction time until reaching the photometric timing T2 to T8 shown in FIG.

  FIG. 18 is a diagram showing an example of a standard calibration curve generated based on the photometric timing shown in FIG. According to the standard calibration curve shown in FIG. 18, since the concentration is steep at a low concentration, the resolution of the calculated concentration is further improved.

  As described above, in the second embodiment, to each of the standard samples S1 to S8, a reagent including an insoluble carrier on which a component that binds to the detection target in the standard samples S1 to S8 is immobilized is added. Each reaction solution in which the reagent is added to the standard samples S1 to S8 is optically measured at different photometric timings. Then, based on the photometric result of each standard sample, a calibration curve is generated from the reaction time until reaching the predetermined absorbance. As a result, regardless of the level of the concentration to be detected, it is possible to uniquely determine the concentration to be detected from the reaction time until reaching the predetermined absorbance.

  The standard calibration curve may be a combination of the correlation between concentration and absorbance and the correlation between concentration and reaction time. The standard calibration curve is represented, for example, by the correlation between concentration and absorbance when the concentration of the detection target is low, and is represented by the correlation between the concentration and the reaction time when the concentration of the detection target is medium to high. Ru. Specifically, for example, as shown in FIG. 19, the response time until reaching the photometric timings T5 to T8 is acquired for standard samples with medium to high concentration. Then, based on the acquired reaction time, a standard calibration curve is generated for a sample whose concentration to be detected is medium to high. In addition, for standard samples with low concentrations, the absorbance is measured from the difference between the absorbance measured in the initial cycle in which the reaction in the reaction solution has not progressed and the absorbance measured at photometric timings T1 to T5 shown in FIG. Calculate Then, based on the calculated absorbance, a standard calibration curve for a sample having a low concentration to be detected is generated.

  FIG. 20 and FIG. 21 show examples of standard calibration curves generated by the method shown in FIG. The standard calibration curve shown in FIG. 20 is represented by the correlation between concentration and absorbance, and is used for measurement of a low concentration detection target. The standard calibration curve shown in FIG. 21 is represented by the correlation between concentration and reaction time, and is used for measurement of medium to high concentration detection targets. This makes it possible to determine the concentration of the detection target from the absorbance for the measurement of the detection target having a low concentration. In addition, for the measurement of medium to high concentration target, the concentration of the detection target can be obtained from the reaction time until the predetermined absorbance is reached. That is, regardless of the level of the concentration to be detected, the concentration to be detected can be uniquely determined.

(Generation of calibration data by automatic analyzer 1)
Next, an example of an operation in which the automatic analyzer 1 generates calibration data will be described.

  First, in the analysis mechanism 2, standard data is generated for at least one of the standard samples S1 to S8 related to a predetermined inspection item. When the standard data is generated, the analysis circuit 3 executes the calibration data generation function 31. When the calibration data generation function 31 is executed, the analysis circuit 3 reads from the storage circuit 8 data on a standard calibration curve for a predetermined inspection item and data on a photometric timing line. The analysis circuit 3 calculates a reaction curve by drawing an approximate curve with respect to the measurement value of the standard sample included in the standard data.

  The analysis circuit 3 determines whether the intersection of the photometric timing line and the response curve is within the maximum number of measurements, for example, 33 cycles. As shown in FIG. 13, when the photometric timing line falls within 33 cycles, the photometric timing line and the response curve always cross within 33 cycles. The analysis circuit 3 calculates the reaction time until reaching the photometric timing which is an intersection point of the response curve and the photometric timing line. The analysis circuit 3 corrects and interpolates, for example, a standard calibration curve set as shown in FIG. 14 based on the reaction time until the photometric timing is reached. In this way, a calibration curve that enables measurement of test data of a predetermined test item is obtained. The analysis circuit 3 stores, for example, the corrected calibration curve in the storage circuit 8 as calibration data.

  As shown in FIG. 15, when the photometric timing line does not fall within 33 cycles, the intersection with the photometric timing line exists in a cycle exceeding 33 cycles depending on the calculated response curve. When the intersection point of the photometric timing line and the reaction curve does not exist within 33 cycles, the analysis circuit 3 performs, for example, linear regression analysis using four points in the observation interval 30 to 33 for the calculated reaction curve. The analysis circuit 3 extends the reaction curve with the approximate straight line calculated by linear regression analysis. The analysis circuit 3 calculates the reaction time until reaching the photometric timing which is an intersection point of the response curve stretched after 33 cycles and the photometric timing line.

  Note that for a reaction curve for which an approximate straight line greater than a predetermined determination coefficient can not be calculated by linear regression analysis, the photometric timing line is preset so that the response curve and the photometric timing line intersect within 33 cycles. I do not care. This makes the calculation of the reaction time more accurate.

  The analysis circuit 3 corrects and interpolates, for example, the standard calibration curve set in FIG. 16 based on the reaction time until reaching the photometric timing. In this way, a calibration curve that enables measurement of test data of a predetermined test item is obtained. The analysis circuit 3 stores, for example, the corrected calibration curve in the storage circuit 8 as calibration data.

  The method of acquiring the calibration curve is not limited to the above. The analysis circuit 3 may acquire a calibration curve without using a standard calibration curve.

  Specifically, for example, measurements on all the standard samples S1 to S8 are performed to generate standard data as absorbance. The analysis circuit 3 calculates a reaction curve by drawing an approximate curve with respect to the measurement values of the standard samples S1 to S8 included in the standard data. The analysis circuit 3 calculates the reaction time until reaching each of the light measurement timings T1 to T8 which is the intersection point of the reaction curve and the light measurement timing line. The analysis circuit 3 obtains a calibration curve based on the reaction time until reaching each of the photometric timings T1 to T8. The analysis circuit 3 stores, for example, a calibration curve in the storage circuit 8 as calibration data.

  As described above, in the second embodiment, the automatic analyzer 1 acquires the intensity curve representing the temporal change of the photometric intensity based on the acquired photometric intensity by the calibration data generation function 31 of the analysis circuit 3, Calibration data is generated based on the reaction time until reaching the intersection of the intensity curve and the photometric timing line. Thus, the automatic analyzer 1 can generate calibration data with a wide dynamic range.

(Generation of analytical data by automatic analyzer 1)
Next, an example of an operation in which the automatic analyzer 1 generates analysis data will be described.

  When the measurement operation is started, test data for a predetermined inspection item is generated by the analysis mechanism 2. When the analysis circuit 3 receives the test data output from the analysis mechanism 2, the analysis circuit 3 executes an analysis data generation function 32. When the analysis data generation function 32 is executed, the analysis circuit 3 reads out from the storage circuit 8 calibration data stored for the inspection item corresponding to the test data and data on the photometric timing line. The analysis circuit 3 calculates a test data approximate curve by drawing an approximate curve with respect to a plurality of actually measured absorbances included in the test data.

  The analysis circuit 3 determines whether the intersection of the photometric timing line and the response curve is within the maximum number of measurements, for example, 33 cycles. FIG. 22 is a diagram illustrating an example of reaction curve C1 to C8, standard light measurement timing lines, and a test data approximate curve calculated based on test data for standard samples S1 to S8. In FIG. 22, the test data approximate curve is represented by a thick two-dot chain line, and the photometric timing line and the response curve intersect within 33 cycles.

  If the intersection of the photometric timing line and the response curve is within 33 cycles, the analysis circuit 3 generates analysis data using photometric timing near the intersection, for example, photometric timings T4 and T5. For example, the analysis circuit 3 connects T4 and T5 on the photometric timing line shown in FIG. 22 with a straight line. Here, assuming that a point at which the photometric timing line intersects the test data approximate curve is X, it is assumed that T4X: XT5 = a: b. The analysis circuit 3 calculates the point at which the line segment T4T5 is divided into a: b on the corrected calibration data on the inspection item corresponding to the test data. This point is a point X where the intersection point X on the photometric timing line is replaced on the calibration data. FIG. 23 is a diagram showing an example of corrected calibration data corresponding to FIG. 22 and the calculated point X. In FIG. 23, a vertical line is drawn from point X to the x axis of the graph, and the value intersecting the x axis is the concentration of the detection target in the sample.

  As shown in FIG. 24, when the photometric timing line does not fit within 33 cycles, the intersection with the photometric timing line exists in a cycle exceeding 33 cycles depending on the test data approximation curve. When the intersection point of the photometric timing line and the test data approximate curve does not exist within 33 cycles, the analysis circuit 3 performs linear regression using, for example, four points of observation intervals 30 to 33 for the calculated test data approximate curve. Perform the analysis. The analysis circuit 3 extends the test data approximate curve with the approximate straight line calculated by linear regression analysis. The analysis circuit 3 obtains an intersection point X of the test data approximate curve extended after 33 cycles and the photometric timing line.

  The analysis circuit 3 generates analysis data using photometric timing near the intersection point, for example, photometric timings T2 and T3. For example, the analysis circuit 3 connects T2 and T3 on the photometric timing line shown in FIG. 24 with a straight line. At this time, it is assumed that T2X: XT3 = a: b. The analysis circuit 3 calculates a point X at which the line segment T2T3 is divided into a: b on the corrected calibration data on the inspection item corresponding to the test data. This point X corresponds to the intersection point X on the photometric timing line. FIG. 25 is a diagram showing an example of the corrected calibration data corresponding to FIG. 24 and the calculated point X. In FIG. 25, a vertical line is drawn from point X to the x axis of the graph, and the value intersecting the x axis is the concentration of the detection target in the sample.

  The method of acquiring the concentration of the detection target in the sample is not limited to the above. For example, the analysis circuit 3 may calculate the reaction time to reach the intersection point X of the photometric timing line and the test data approximate curve. The analysis circuit 3 compares the calculated reaction time with, for example, the corrected calibration data shown in FIG. 23, and calculates the concentration of the detection target in the sample.

  Further, depending on the concentration of the detection target in the sample, for example, the photometric values in the observation sections 30 to 33 may vary. In such a case, the accuracy of the approximate straight line calculated by linear regression analysis using four points in the observation intervals 30 to 33 is not high. The analysis circuit 3 may set an identifier indicating that the accuracy of the calculated approximate straight line is not high, for example, an error flag, for example, when the variation of the photometric value in the observation sections 30 to 33 is equal to or greater than a preset value. .

  As described above, in the second embodiment, the automatic analyzer 1 uses the analysis data generation function 32 of the analysis circuit 3 to generate an intensity curve representing the change over time of the photometric intensity based on the acquired photometric intensity of the test sample. To get Then, the automatic analyzer 1 generates analysis data based on the reaction time until reaching the intersection point of the acquired intensity curve and the photometric timing line. By the way, when measuring the concentration of the detection target based on the calibration curve generated using the absorbance at the intersection of the intensity curve and the photometric timing line, minute variations in absorbance at the time of measurement greatly affect the measurement result There is. This is particularly pronounced at low concentrations. On the other hand, the calibration curve generated using the reaction time to reach the intersection of the intensity curve and the photometric timing line has high resolution even at low concentrations. For this reason, even if the reaction time at the time of measurement has a slight variation, the measurement result hardly varies greatly.

  According to at least one embodiment described above, in the measurement using the insoluble carrier on which the antibody or the antigen is immobilized, the detection target can be measured with high sensitivity and high concentration, that is, high sensitivity and wide dynamic range Can be realized.

  The word “processor” used in the description of the embodiment is, for example, a central processing unit (CPU), a graphics processing unit (GPU), or an application specific integrated circuit (ASIC)), a programmable logic device It means circuits such as (for example, Simple Programmable Logic Device (SPLD), Complex Programmable Logic Device (CPLD), and Field Programmable Gate Array (FPGA)). The processor realizes a function by reading and executing a program stored in the memory circuit 8. Instead of storing the program in the memory circuit 8, the program may be directly incorporated in the circuit of the processor. In this case, the processor implements the function by reading and executing a program embedded in the circuit. Each processor in each of the above embodiments is not limited to the case where it is configured as a single circuit for each processor, but a plurality of independent circuits are combined to configure as one processor, and the functions thereof are realized It is also good. Furthermore, a plurality of components in the above embodiment may be integrated into one processor to realize its function.

  While certain embodiments of the present invention have been described, these embodiments have been presented by way of example only, and are not intended to limit the scope of the invention. These embodiments can be implemented in other various forms, and various omissions, replacements, and modifications can be made without departing from the scope of the invention. These embodiments and modifications thereof are included in the invention described in the claims and the equivalents thereof as well as included in the scope and the gist of the invention.

DESCRIPTION OF SYMBOLS 1 automatic analysis device 2 analysis mechanism 3 analysis circuit 4 drive mechanism 5 input interface 6 output interface 7 communication interface 8 storage circuit 9 control circuit 31 calibration data generation function 32 analysis data generation function 91 ... System control function 92 ... Calibration control function 93 ... Measurement control function 201 ... Reaction disk 202 ... Constant temperature section 203 ... Sample disk 204 ... Reagent storage 205 ... Reagent storage 206 ... Sample dispensing arm 207 ... Sample dispensing probe 208 ... Reagent portion Note arm 209 ... reagent dispensing probe 210 ... reagent dispensing arm 211 ... reagent dispensing probe 212 ... agitation unit 213 ... agitation unit 214 ... photometric unit 215 ... cleaning unit 2011, 2011-1 to 2011-8 ... reaction container 2041 ... Reagent rack 2051 ... Reagent rack 2121 ... Stirring arm 131 ... stirring arm

Claims (20)

A calibration curve generation method for generating a calibration curve based on photometric results of a plurality of standard samples corresponding to different concentrations, including detection targets of known concentrations,
A calibration curve generation method for generating the calibration curve based on photometric results of a plurality of standard samples acquired at different photometric timings.   The calibration curve generation method according to claim 1, wherein the photometric timing is set so as to be stepwise delayed as the concentration of the standard sample changes from high concentration to low concentration.   The method according to claim 1 or 2, wherein the calibration curve represents a correlation between reaction time and concentration. Based on the intensity curve of each of the plurality of standard samples, a photometric timing line is set which is different at the time of acquiring the photometric intensity for each of the standard samples,
The calibration curve generation method according to claim 1, wherein an intersection point of an intensity curve for each of the standard samples and the light measurement timing line is used as the light measurement timing.   Based on the intensity curve of each of the plurality of standard samples, the timing of acquiring the photometric intensity for each of the standard samples is gradually delayed based on the intensity curve of each of the plurality of standard samples, from the high concentration standard sample to the low concentration standard sample. The method of generating a calibration curve according to claim 4 to be set.   The calibration curve generation method according to claim 4 or 5, wherein the photometric timing line is set by regression analysis using an intensity curve for each of the standard samples.   5. The method of generating a calibration curve according to claim 4, wherein the photometric timing line is set to intersect an actual measurement point included in an intensity curve of each of the standard samples.   The calibration curve generation method according to any one of claims 4 to 7, wherein a plurality of photometric timing lines are set.   The calibration curve generation method according to any one of claims 1 to 8, wherein the calibration curve is generated based on the photometric intensity acquired at the photometric timing.   The calibration curve generation method according to any one of claims 1 to 8, wherein the calibration curve is generated based on a reaction time acquired at the photometric timing.   The calibration curve is generated based on the reaction time acquired at the photometric timing according to the concentration of the standard sample, or the calibration curve is generated based on the photometric intensity acquired at the photometric timing. The calibration curve generation method as described in any one of 8. Approximating and stretching at least one of the intensity curves,
5. The calibration curve generation method according to claim 4, wherein an intersection point between the stretched intensity curve and the photometric timing line is used as the photometric timing. An ejection unit for adding a reagent containing an insoluble carrier on which a component that binds to the detection target is immobilized to a standard sample containing the detection target at a known concentration;
A photometry unit that optically measures a reaction solution in which the reagent is added to the standard sample;
A storage unit for storing information on photometric timing which is set to be different for each of a plurality of standard samples different in concentration of a detection target included;
An automatic analyzer comprising: a calibration data generation unit configured to generate calibration data based on the photometry result acquired at the photometry timing.   The automatic analyzer according to claim 13, wherein the photometric timing is set so as to be gradually delayed as the concentration of the standard sample changes from high concentration to low concentration.   The automatic analyzer according to claim 13 or 14, wherein the calibration data represents a correlation between reaction time and concentration. The photometric unit obtains the photometric intensity of the standard sample by optically measuring the reaction solution in which the reagent is added to the standard sample.
The storage unit stores a photometric timing line which is set such that the point of time when the information on the acquired photometric intensity is acquired is different for each of a plurality of standard samples having different concentrations of the detection target.
The calibration data generation unit acquires, based on the acquired photometric intensity, an intensity curve representing a temporal change of the photometric intensity, and uses an intersection point of the intensity curve and the photometric timing line as the photometric timing. The automatic analyzer according to 14.   The photometric timing line is based on the intensity curve of each of the plurality of standard samples so that the point at which the photometric intensity is obtained for each of the standard samples is gradually delayed from the high density standard sample to the low density standard sample. The automatic analyzer according to claim 16, which is set.   The automatic analyzer according to claim 16 or 17, wherein the calibration data generation unit generates the calibration data based on the photometric intensity acquired at the photometric timing.   The automatic analyzer according to claim 16 or 17, wherein the calibration data generation unit generates the calibration data based on a reaction time acquired at the light measurement timing. The discharge unit adds the reagent to a test sample,
The photometry unit optically measures the reaction solution in which the reagent is added to the test sample to obtain the photometric intensity of the test sample.
Based on the acquired photometric intensity of the test sample, an intensity curve representing a change with time of the photometric intensity is acquired, and information acquired at the intersection of the intensity curve and the photometric timing line, and the calibration data The automatic analyzer according to any one of claims 16 to 19, further comprising an analysis data generation unit that generates analysis data.