JP6300676B2 - Analysis method and automatic analyzer - Google Patents

Description

  The present invention relates to an analysis method and an automatic analyzer that perform analysis using an antigen-antibody reaction using blood, serum, plasma, or the like as a sample or specimen.

  For analysis methods using blood, serum, plasma, or body fluid as a sample or specimen, a trace substance such as a hormone, tumor marker, infectious disease pathogen marker, or infectious antibody in a sample is utilized using binding based on an antigen-antibody reaction. A method for detecting qualitatively or quantitatively is known. However, in the analysis method using the antigen-antibody reaction, when the sample contains a high concentration of the substance to be analyzed, the numerical value of the analysis result becomes lower than the actual value (zone phenomenon or hook phenomenon) May occur).

  With regard to such a zone phenomenon, for example, Patent Document 1 (Japanese Patent Publication No. 2010-520461) discloses an improved and accurate quantitative analysis of the analyte concentration by reducing the influence of immunoassay interference such as the prozone phenomenon. For the purpose of enabling measurement, the target sample present in the test sample is captured by the capture antibody, a complex of the capture antibody, the sample and the first antibody conjugate is formed, and the unbound sample is removed from the mixture. A technique relating to an immunoassay in which an analyte is detected by an antibody conjugate by adding a second antibody conjugate that binds to the analyte later is disclosed.

Special table 2010-520461

  By the way, a so-called antigen-antibody method two-step reaction (two-step method) is known as a method aimed at avoiding the zone phenomenon and the hook phenomenon. In general, in a two-step reaction, a solid phase and a capture antibody or a capture antigen and a sample are mixed, and after forming a conjugate of an analyte and a capture antibody, washing is performed, and unbound sample components and analysis are performed. After removing the target (bound / free separation) and adding a labeled antibody to form a capture antibody-analyte-labeled antibody sandwich, washing is performed to detect luminescence, etc. derived from the labeled antibody, and to be analyzed Measure quantity or concentration. In this two-step reaction, the unbound analyte is removed by the first washing, so that inhibition of the binding of the labeled antibody is suppressed, and therefore the zone phenomenon is unlikely to occur.

  In addition, as a method for suppressing the analysis time and the labor of the operator, the solid phase, the capture antibody, the capture antigen, the labeled antibody, the labeled antigen and the sample are dispensed after the sample is dispensed and before the labeled antibody is dispensed. There is known a one-step reaction (one-step method) in which a mixing reaction is performed without performing washing, and then washing is performed to remove unbound reagents and sample components on the solid phase and measurement of an analysis object is performed. However, in the one-step reaction, the zone phenomenon greatly affects the measurement result. In the one-step method, if an analyte is contained in a high concentration in the sample, or if it is expected, the sample is diluted in advance, and the sample is diluted to a concentration at which the zone phenomenon is not expected to occur. However, in this case, it is often necessary to perform two measurements for each sample: measurement without sample dilution and measurement with the sample.

  The present invention has been made in view of the above, and an object of the present invention is to provide an analysis method and an automatic analyzer that can suppress a decrease in accuracy of measurement results due to a zone phenomenon.

  In order to achieve the above object, the present invention includes a procedure for performing a first capturing body addition process of adding a capturing antibody reagent or a capturing antigen reagent containing a capturing body to a reaction container in which a sample to be analyzed is stored, A procedure for performing a first labeled body addition process in which a labeled antigen reagent or labeled antibody reagent containing a labeled body is added to a reaction container, and a first reaction solid phase in which a reaction solid phase reagent including a reaction solid phase is added to the reaction container. A procedure for performing a phase addition treatment, a procedure for performing a first reaction solid phase washing treatment for washing the reaction solid phase in order to remove components other than the components that form bonds including the reaction solid phase, and the reaction Removing at least a part of the reaction solid phase from the container, and performing a first measurement process for measuring a physical quantity related to the labeled body including the reaction solid phase to form a bond; and after the first measurement process Second time in the reaction vessel A procedure for performing a label addition process, a procedure for performing a second reaction solid phase washing process after the second label addition process, and a procedure for performing a second measurement process after the second reaction solid phase washing process And a procedure for selectively determining one measurement result from a plurality of measurement result candidate values obtained by the first measurement process based on the result of the second measurement process.

  A decrease in the accuracy of the measurement result due to the zone phenomenon can be suppressed.

It is a figure which shows schematically the whole structure of an automatic analyzer. It is a figure which extracts and shows a sample dispensing mechanism with a periphery structure. It is a figure which shows the flow of an analysis process roughly. It is a figure which shows the relationship between the signal value and the density | concentration of an analyte in an example of the calibration curve obtained in the measurement process (1st time) of the process 3 of FIG. It is a figure which shows an example of the signal value obtained in the measurement process (2nd time) of the process 7 of FIG. It is a flowchart which shows the processing content of an analysis process.

  Hereinafter, an embodiment of the present invention will be described with reference to FIGS.

(1) Overall Configuration FIG. 1 is a diagram schematically showing an overall configuration of an automatic analyzer according to the present embodiment.

  In FIG. 1, an automatic analyzer 100 includes a sample container 108 for storing a sample or specimen (hereinafter simply referred to as a sample) to be analyzed, a rack transport line 113 for transporting a rack 107 on which the sample container 108 is mounted, and a sample. A labeled reagent container 201A containing a labeled antigen or a labeled reagent containing a labeled antibody that forms a specific antigen-antibody bond with the analyte component in the sample, and a capture that forms a specific antigen-antibody bond with the analyte component in the sample A reaction solid phase reagent container containing a reaction solid phase reagent containing a capture reagent container 201B containing a capture reagent containing an antigen or a capture antibody and a reaction solid phase that forms a specific antigen-antibody bond with the analyte in the sample Reaction in which 201C, reagent disk 202 carrying reagent containers 201A, 201B, and 201C, and a sample, a labeled reagent, and a reaction solid phase reagent are dispensed and reacted. Container 205, reaction disk 203 carrying reaction container 205, sample dispensing mechanism 206 for dispensing a sample from sample container 108 to reaction container 205, labeling reagent container 201A, capture reagent container 201B, and reaction solid phase reagent At the reagent dispensing mechanism 208 for dispensing the labeling reagent, capture reagent and reaction solid phase reagent from the container 201C to the reaction container 205, and at the reaction liquid suction position 212 of the reaction disk 203, the reaction liquid in the reaction container 205 is sucked, A detection unit 215 (analysis mechanism) that performs qualitative and quantitative analysis of components to be analyzed based on physical quantities of various labeling substances (labeled substances) in the reaction solution, a sample dispensed in the reaction vessel 205, a labeling reagent, and a reaction A reaction solid phase assembly mechanism 402 that collects the reaction solid phase in the mixed solution of the solid phase reagent on the inner wall of the reaction vessel, an unused reaction vessel 205, and a dispensing tip 210. A transfer mechanism 216 for transferring, is schematically a control unit 223 which controls the automatic analyzer 100 overall operation performs the process of the analysis process or the like according to the analysis of the sample (described below).

(1-1) Rack transport line 113
A plurality of sample containers 108 are transported along the rack transport line 113 in a state of being mounted on the rack 107. The sample container 108 contains a biological sample to be analyzed (hereinafter simply referred to as a sample) such as serum, plasma, and body fluid. A sample suction position 207 is disposed on the rack transport line 113.

(1-2) Reagent disc 202
On the reagent disk 202, a plurality of reagent containers 201A, 201B, and 201C in which various reagents used for analysis processing are stored are arranged in the circumferential direction. The reagent disk 202 is rotationally driven in the circumferential direction by a rotation driving device (not shown), thereby transporting the reagent containers 201A, 201B, 201C in the circumferential direction. A reagent suction position 209 is disposed on the transport path of the reagent containers 201A, 201B, and 201C in the reagent disk 202.

  Examples of the reagent container disposed on the reagent disk 202 include a labeling reagent container 201A, a capture reagent container 201B, and a reaction solid phase reagent container 201C. In the labeling reagent container 201A, an antibody or antigen that forms a specific antigen-antibody bond with a component to be analyzed in a sample is labeled with a labeled antibody or a labeled reagent containing a labeled antigen (labeled antibody reagent or labeled antigen reagent). ) And a capture reagent (capture antigen reagent or capture antibody) containing a capture body (capture antibody or labeled antigen) that forms a specific antigen-antibody bond with the analysis target component in the sample. Reagent) and a reaction solid phase reagent container 201C contains a reaction solid phase reagent including a reaction solid phase.

  As another aspect of the reagent container disposed on the reagent disk 202, a reagent having the functions of two types of reagents among the labeling reagent, the capture reagent, and the reaction solid phase reagent (for example, two kinds of reagents are mixed). A reagent container containing a reagent and a reagent container containing a reagent having the function of the remaining one type of reagent, or a reagent having the functions of three types of reagents simultaneously (for example, three types of reagents) There may be a case where a reagent container containing a mixed reagent) is disposed.

  In the automatic analyzer according to the present embodiment, when measuring analysis of trace substances such as hormones, tumor markers, infectious disease pathogen markers, and infectious antibodies using blood, serum, plasma or body fluid as a sample, the protein to be analyzed For each analysis item, protein or the like in the sample is detected qualitatively or quantitatively by binding based on the antigen-antibody reaction. As the reaction solid phase surface for the antigen-antibody reaction, plastic, glass plate or particles, magnetic particles containing iron oxide, or the like are used. Such an antibody or antigen that binds to the protein to be analyzed in the solid phase is referred to as a capture antibody, a capture antigen, a capture antibody, or a capture antigen. In addition, antibodies or antigens that bind radioisotopes, fluorescent dyes, luminescent dyes, enzymes, rare earth complexes, metal ions, etc., and bind to proteins to be analyzed are labeled antigens, labeled antibodies, labeled antigens, conjugates, tracers, etc. Called.

(1-3) Reaction disk 203
On the reaction disk 203, a plurality of reaction containers 205 that contain a mixed solution (reaction solution) of a sample and various reagents are arranged side by side in the circumferential direction and held at a constant temperature. The reaction disk 203 is driven to rotate in the circumferential direction by a rotation drive device (not shown), thereby conveying the reaction container 205 in the circumferential direction. On the transport path of the reaction container 205 on the reaction disk 203, a reaction container installation position 204, a sample discharge position 221, a reagent addition position 222, a reaction liquid suction position 212, and the like are arranged. The reaction disk 203 is provided with a reaction solid phase assembly mechanism 402 (see FIG. 2 and the like later) for collecting the reaction solid phase in the mixed solution (reaction solution) in the reaction vessel 205 on the inner wall of the reaction vessel 205. ing.

(1-3.1) Reaction solid phase assembly mechanism 402
The reaction solid phase assembly mechanism 402 collects and releases the reaction solid phase in accordance with the execution of various processes by the control unit 223, and the reaction solid phase in the mixed solution (reaction solution) contained in the reaction vessel 205. Are assembled and fixed to the inner wall of the reaction vessel 205. With the reaction solid phase fixed to the inner wall of the reaction vessel 205, pure water, a buffer solution, a dedicated cleaning solution, etc. are added to and removed from the reaction vessel 205 without causing the reaction solid phase to flow out. Washing (reaction solid phase washing treatment described later) can be performed. As a specific configuration of the reaction solid phase assembly mechanism 402, for example, when the reaction solid phase includes a magnetic material, the reaction solid phase is attracted to the inner wall of the reaction vessel 205 by magnetism such as a magnet to be fixed.

(1-4) Sample dispensing mechanism 206
FIG. 2 is a diagram showing the sample dispensing mechanism 206 extracted along with the peripheral configuration.

  The sample dispensing mechanism 206 is configured to be horizontally rotatable and vertically movable, and at the sample suction position 207, the tip of the probe 401 is brought into contact with the sample in the sample container 108 to perform a predetermined amount of suction. Then, the sample is discharged into the reaction container 205 at the sample discharge position 221. On the trajectory along which the probe 401 of the sample dispensing mechanism 206 moves, a coupling position 218 for coupling an unused dispensing tip 210 used for dispensing to the probe 401 and a tip discarding position for discarding a used dispensing tip. 220 is arranged. After dispensing, the probe 401 is cleaned of sample components by a probe cleaner 403. In the probe cleaner 403, the outer surface of the probe 401 inserted into the cleaning cup 407 is cleaned with the cleaning water supplied to the cleaning cup 407 through the water supply valve 408, and the cleaning water in the cleaning cup 407 is sucked up by the probe 401. Then, the probe 401 is cleaned by moving to the waste bottle 404 and discharging.

(1-5) Reagent dispensing mechanism 208
The reagent dispensing mechanism 208 has the same configuration as the sample dispensing mechanism 206. That is, the reagent dispensing mechanism 208 is configured rotated and vertically movably in the horizontal direction, the reagent containers 201A to the tip of the probe in the reagent aspirating position 209, 201B, by wetted the reagent in 201C Tokoro A fixed amount is aspirated and discharged into the reaction container 205 at the reagent addition position 222. After the dispensing, the reagent components are washed by a probe washing machine having the same configuration as the probe washing machine 403 in the sample dispensing mechanism 206. That is, in the probe washing machine of the reagent dispensing mechanism 208, the washing water supplied to the washing cup through the water supply valve is used to wash the outer surface of the probe inserted into the washing cup, and the probe is used to wash the washing water in the washing cup. The probe is washed by sucking up and moving to a waste bottle and discharging.

(1-6) Transfer mechanism 216
The automatic analyzer 100 includes a reaction container storage unit 219 that stores unused reaction containers 205 and a dispensing tip storage unit 217 that stores unused dispensing tips 210. The tip 210 is transferred by the transfer mechanism 216. The transfer mechanism 216 is configured to be movable in three directions of the X axis, the Y axis, and the Z axis, and can transport the reaction container 205 from the reaction container storage unit 219 to the reaction container installation position 204 of the reaction disk 203, The dispensing tip 210 is transferred from the dispensing tip storage unit 217 to the tip coupling position 218.

(1-7) Detection unit 215
The detection unit 215 is provided with a reaction liquid suction mechanism 211 that sucks the reaction liquid in the reaction vessel 205 at a reaction liquid suction position 212 of the reaction disk 203 and sends it to a flow cell (not shown) arranged inside the detection unit. The physical quantity relating to the labeled antigen or the labeled antibody in the reaction liquid, that is, the physical quantity relating to the labeled body is measured by passing the reaction liquid through the flow cell. As a physical quantity to be measured, for example, when a fluorescent dye or a luminescent dye is used for the label, the light amount is measured, and when a radioisotope is used for the label, the radiation is measured.

  The reaction solution suction mechanism 211 is configured to be able to rotate in the horizontal direction and move up and down, and a buffer for sucking a buffer solution (described later) used for analysis on the trajectory on which the probe of the reaction solution suction mechanism 211 moves. A liquid suction position 213 and a cleaning liquid suction position 214 for sucking a cleaning liquid used for cleaning the flow path of the sucked reaction liquid are arranged. In the reaction solution suction mechanism 211, the outer wall of the probe after suction of the reaction solution and the buffer solution is washed by a probe washing machine (not shown).

(1-8) Control unit 223
The control unit 223 controls the entire operation of the automatic analyzer 100, and as a process related to the analysis of the sample, a labeled body addition process, a capturing body addition process, a reaction solid phase addition process, a reaction solid phase cleaning process, and a measurement process It has a function as an analysis processing control apparatus for performing the above.

(1-8.1) Labeled body addition process In the labeled body addition process, the labeled reagent (labeled antibody reagent or labeled antigen reagent) accommodated in the labeled reagent container 201A is dispensed into the reaction container 205 by the reagent dispensing mechanism 208. In this process, either a labeled antibody reagent or a labeled antigen reagent is selected and used depending on the component to be analyzed.

(1-8.2) Capturing body addition process In the capturing body addition process, the capture reagent (capture antibody reagent or capture antigen reagent) accommodated in the capture reagent container 201B is dispensed into the reaction container 205 by the reagent dispensing mechanism 208. Either capture antibody reagent or capture antigen reagent is selected and used depending on the component to be analyzed.

(1-8.3) Reaction Solid Phase Addition Process The reaction solid phase addition process is a process of dispensing the reaction solid phase reagent accommodated in the reaction solid phase reagent container 201C into the reaction container 205 by the reagent dispensing mechanism 208. .

  The labeling body addition process, the capturing body addition process, and the reaction solid phase addition process may be performed substantially simultaneously depending on the state of the reagent container disposed on the reagent disk 202. For example, a reagent container containing a reagent having two types of reagent functions simultaneously among a labeling reagent, a capture reagent, and a reaction solid phase reagent and a reagent container containing a reagent having the function of the remaining one kind of reagent are used as a reagent disk. In the case of being arranged in 202, two of the three processes are performed at the same time, and a reagent container containing reagents having the functions of the three types of reagents is arranged in the reagent disk 202. In some cases, three processes are performed simultaneously.

(1-8.4) Reaction solid phase cleaning treatment In the reaction solid phase washing treatment, the reaction solid phase in the reaction solution contained in the reaction vessel 205 is assembled and fixed on the inner wall of the reaction vessel 205 by the reaction solid phase assembly mechanism 402. In this state, the reaction solid phase is washed by adding and removing pure water, a buffer solution, or a dedicated washing solution by the sample dispensing mechanism 206, the reagent dispensing mechanism 208, or a dedicated addition mechanism (not shown). It is a process to perform. By this reaction solid phase washing treatment, components (sample component, reagent component, labeled antibody, capture antibody) other than the component that forms a bond including the reaction solid phase are discarded.

(1-8.5) Measurement Process In the measurement process, the reaction liquid in the reaction vessel 205 is sucked by the reaction liquid suction mechanism 211, and the labeled antigen or the label in the reaction liquid is detected by the flow cell installed in the detection unit 215. This is a process for measuring a physical quantity related to an antibody, that is, a physical quantity related to a label. As a physical quantity to be measured, for example, when a fluorescent dye or a luminescent dye is used for the label, the light amount is measured, and when a radioisotope is used for the label, the radiation is measured. Depending on the type of label, radioimmunoassay, enzyme immunoassay, fluorescent immunoassay, chemiluminescent immunoassay, electrochemiluminescent immunoassay, and the like are known.

(2) Analysis process The analysis process by the control part (analysis process control apparatus) of this Embodiment is demonstrated, referring FIGS. 3-6.

  FIG. 3 is a diagram schematically showing the flow of analysis processing in the present embodiment.

  In the analysis process, first, a capture body addition process in which a capture antibody reagent or a capture antigen reagent is added to a reaction container 205 containing a sample to be analyzed, and a labeled antigen reagent or a labeled antibody reagent are added to the reaction container. The first labeled body addition process and the first reaction solid phase addition process for adding the reaction solid phase reagent to the reaction vessel are performed, followed by stirring and incubation (step 1 in FIG. 3).

  Next, a reaction solid phase cleaning process (first time) is performed in which components other than the components including the reaction solid phase that have formed a bond are removed (that is, bounce-free separation) (step 2 in FIG. 3).

  Next, a measurement process (first time) for measuring a physical quantity related to the label in the reaction container 205 is performed (step 3 in FIG. 3).

  Next, a small amount of labeled substance addition treatment (second time) is performed on the reaction solution remaining after the measurement treatment (first time) (step 4 in FIG. 3). Note that the amount of labeling reagent added (small amount) in the labeling process in the step is such that 1/2 to 1/1000 of the labeling substance is added compared to the amount of the labeling substance added in step 1. Amount.

  Next, the reaction solution subjected to the labeling treatment (second time) is stirred and incubated (step 5 in FIG. 3).

  Next, a reaction solid phase washing process (second time) is performed after the labeling substance addition process (second time) (step 6 in FIG. 3).

  Next, a measurement process (second time) is performed after the reaction solid phase cleaning process (second time) (step 7 in FIG. 3), and a plurality of measurements obtained by the measurement process (first time) based on the measurement result One measurement result is selectively determined from the result candidate values.

  FIG. 4 is a diagram showing the relationship between the signal value and the concentration of the analyte in an example of a calibration curve obtained in the measurement process (first time) of step 3 in FIG.

  In FIG. 4, the calibration curve 40 increases in signal value as the analyte concentration in the sample increases, draws a gentle S-shape, and reaches the maximum value (peak value) of the signal. Further, when the analyte concentration is higher, the signal value changes and gradually decreases as the concentration increases. When there is a drop from the peak value, it can be seen that there is a zone phenomenon. When an unknown sample is measured, the analyte concentration cannot be predicted, so it is unknown whether the concentration is indicative of a zone phenomenon. Therefore, when the measurement result takes the signal value S0, it is on the calibration curve 240. The concentration value VA (state A) or the concentration value VB (state B) in the range where the calibration curve 240 descends due to the occurrence of the zone phenomenon is taken.

  FIG. 5 is a diagram illustrating an example of signal values obtained in the measurement process (second time) of step 7 in FIG. 3.

  In FIG. 5, in the case where the concentration of the analyte in the sample is relatively low (that is, state A), the signal value (T2) after the labeling reagent addition process (small amount) in step 4 of FIG. S1) does not increase significantly compared to the signal value (S1, T1) of the measurement process (first time) in step 3. On the other hand, when the concentration of the analyte in the sample is relatively high (ie, state B), it is captured on the surface of the reaction solid phase at the time of the measurement process (first time) (T1) in step 3 of FIG. Many binding substances remain between the body and the analyte, and therefore, after the labeling reagent addition process (small amount) in step 4 of FIG. 3 (T2), the sandwich of the capture body-antigen (antibody) -labeled body And the signal value (S2) rises because a large number of labeled bodies are observed. That is, when a high signal value (S2) is obtained in the measurement process (second time), it is in state B in FIG. 4, and therefore the concentration value VB is selected and determined as the measurement result. The determination of the state is performed by setting a reference value (Sth) in FIG. 5 in advance and comparing it with the measurement result of the measurement process (second time) to determine whether the analyte concentration is the value of state A (VA) or state B. It is possible to determine whether the value is VB. This reference value (Sth) is for determining whether or not there is an increase in the signal value after the labeling agent addition process in Step 4 of FIG. 3, and is therefore based on the signal value in the measurement process (first time). Thus, the reference value (Sth) itself goes up and down.

  FIG. 6 is a flowchart showing the processing contents of the analysis processing.

  In FIG. 6, a control unit 223 as an analysis control device first dispenses a sample to be analyzed into a reaction container 205 (step S100), and then adds a labeling reagent, a capture reagent, and a reaction solid phase reagent. An addition process is performed (step S110). Subsequently, the mixed solution (reaction solution) in the reaction vessel 205 is stirred and mounted in an incubator to react (step S120). After the reaction is completed, the reaction solution containing the sample and the remaining reagents is removed and the reaction solid phase is removed. A reaction solid phase cleaning process is performed (step S130). Next, the reaction solid phase is suspended in the solution, and the physical quantity (measurement process (first time)) related to the label held on the solid phase via the antigen-antibody binding is measured (step S140). Then, based on the measurement result of the measurement process (first time), a measurement result candidate value (VA) is calculated (step S141), and a measurement result candidate value (VB) is calculated (step S142). In parallel with the measurement process (first time) in step S140 (or when steps S141 and S142 are completed), the labeling reagent is added to the remaining reaction solution in the measurement process (first time) stored in the reaction vessel 205. A labeling reagent addition process (a small amount) is added (step S150). Subsequently, the mixed solution (reaction solution) in the reaction vessel 205 is stirred and loaded into the incubator to react (step S155), and after loaded into the incubator and the reaction is completed, the reaction solid phase washing process (second time) ) (Step S160), and subsequently, a measurement process (second time) is performed (step S170). Here, it is determined whether or not the measurement result (signal value) in step S170 is greater than or equal to the reference value (Sth) (step S180). If the determination result is NO, it is determined that there is no zone phenomenon. The measurement result candidate value (VA) is selected and determined as the measurement result (step S181), and the number of processes is counted. If the determination result in step S180 is YES, it is determined that there is a zone phenomenon, the measurement result candidate (VB) is selected and determined as the measurement result (step S182), and the process ends.

  The effect of the present embodiment configured as described above will be described.

  For analysis methods using blood, serum, plasma, or body fluid as a sample or specimen, a trace substance such as a hormone, tumor marker, infectious disease pathogen marker, or infectious antibody in a sample is utilized using binding based on an antigen-antibody reaction. A method for detecting qualitatively or quantitatively is known. However, in the analysis method using the antigen-antibody reaction, when the sample contains a high concentration of the substance to be analyzed, the numerical value of the analysis result becomes lower than the actual value (zone phenomenon or hook phenomenon) May occur).

  On the other hand, in the present embodiment, the procedure for performing the first capture body addition process of adding the capture antibody reagent or capture antigen reagent containing the capture body to the reaction container containing the sample to be analyzed, A procedure for performing a first labeled body addition process in which a labeled antigen reagent or labeled antibody reagent containing a labeled body is added to a reaction container, and a first reaction solid phase in which a reaction solid phase reagent including a reaction solid phase is added to the reaction container. A procedure for performing a phase addition treatment, a procedure for performing a first reaction solid phase washing treatment for washing the reaction solid phase in order to remove components other than the components that form bonds including the reaction solid phase, and the reaction A procedure for performing a first measurement process for measuring a physical quantity related to a label in a container, a procedure for performing a second label addition process after the first measurement process, and a procedure for performing a second label addition process after the second measurement process. The second reaction solid phase cleaning process And a plurality of measurement results obtained by the first measurement process based on the result of the second measurement process and the procedure of performing the second measurement process after the second reaction solid phase cleaning process. And a procedure for selectively determining one measurement result from the candidate value, it is possible to suppress a decrease in accuracy of the measurement result due to the zone phenomenon.

  That is, when performing an analytical measurement based on an antigen-antibody reaction of a sample in an analysis method or apparatus based on a fluorescence measurement method or a luminescence measurement method, there is a factor that decreases the signal value due to the zone phenomenon and impairs the accuracy of the measurement value. In response to this, the amount and concentration of the analyte in the sample can be correctly measured.

  Furthermore, the true value can be measured correctly even if the measurement object in the sample has a high concentration that causes the zone phenomenon. Similarly, if it is a negative value, it can be determined that the negative value is correct.

  In addition, when the sample is diluted and remeasurement is required, the reagent amount twice as large as that in the normal measurement is required. However, in this embodiment, it is sufficient that the labeling reagent is about 1/2 to 1/1000. In particular, the amount and concentration of the analyte in the sample can be measured correctly. Further, it is not necessary to repeat the two tests of the main measurement and the dilution measurement, and therefore the time required for the measurement can be shortened.

  Furthermore, although the one-step method has a problem that the zone phenomenon greatly affects the measurement result, in the present embodiment, the number of measurement range digits can be doubled while covering the weak points. . For example, in the case of having a measurement range of 10 ^ 4, it is possible to effectively measure up to a range of 10 ^ 8, and infectious disease items can be covered with the minimum reagent usage.

  In the present embodiment, the reference value is set based on the measurement result of the measurement process (first time), and the measurement process (first time) is compared with the measurement result of the measurement process (second time) and the reference value. However, the measurement result (see FIG. 5 and the like) is selectively limited (see FIG. 5 and the like). That is, for example, at least two measurements are performed immediately after the addition of the labeling reagent in the measurement process (second time) and after a predetermined time has elapsed since the addition of the labeling reagent, and based on the difference between the measurement results, The first measurement result may be selectively determined. Even in this case, the same effect as the present embodiment can be obtained.

  In the present embodiment, the reaction solid phase cleaning process (second time) is performed. However, the present invention is not limited to this, and the measurement process (second time) is performed after the label addition process (second time). , May be configured.

100 automatic analyzer 107 rack 108 sample container 113 rack transport line 201A labeling reagent container 201B capture reagent container 201C reaction solid phase reagent container 203 reaction disk 204 reaction container installation position 205 reaction container 206 sample dispensing mechanism 207 sample suction position 208 reagent part Injection mechanism 209 Reagent suction position 210 Dispensing chip 211 Reaction liquid suction mechanism 212 Reaction liquid suction position 213 Buffer liquid suction position 214 Cleaning liquid suction position 215 Detection unit 216 Reaction container transfer mechanism 217 Dispensing chip storage unit 218 Chip coupling position 219 Reaction container Storage unit 220 Chip discarding position 221 Sample discharge position 222 Reagent addition position 223 Control unit 240 Calibration curve 402 Reaction solid phase assembly mechanism

Claims (6)

A procedure for performing a first capture body addition process of adding a capture antibody reagent or capture antigen reagent containing a capture body to a reaction container containing a sample to be analyzed;
A procedure for performing a first labeled body addition process of adding a labeled antigen reagent or labeled antibody reagent containing a labeled body to the reaction container;
A procedure for performing a first reaction solid phase addition treatment in which a reaction solid phase reagent containing a reaction solid phase is added to the reaction vessel;
A procedure for performing a first reaction solid phase washing treatment for washing the reaction solid phase in order to remove components other than the component that forms a bond including the reaction solid phase;
Removing at least a part of the reaction solid phase from the reaction vessel, and performing a first measurement process for measuring a physical quantity related to the labeled body including the reaction solid phase to form a bond;
A procedure for performing a second labeled body addition treatment on the reaction vessel after the first measurement treatment;
A procedure for performing a second reaction solid phase washing treatment after the second labeling agent addition treatment;
A procedure for performing a second measurement process after the second reaction solid phase washing process;
And a procedure for selectively determining one measurement result from a plurality of measurement result candidate values obtained by the first measurement process based on the result of the second measurement process. A procedure for performing a first capture body addition process of adding a capture antibody reagent or capture antigen reagent containing a capture body to a reaction container containing a sample to be analyzed;
A procedure for performing a first labeled body addition process of adding a labeled antigen reagent or labeled antibody reagent containing a labeled body to the reaction container;
A procedure for performing a first reaction solid phase addition treatment in which a reaction solid phase reagent containing a reaction solid phase is added to the reaction vessel;
Removing at least a part of the reaction solid phase from the reaction vessel, and performing a first measurement process for measuring a physical quantity related to the labeled body including the reaction solid phase to form a bond;
A procedure for performing a second labeled body addition treatment on the reaction vessel after the first measurement treatment;
A procedure for performing a second measurement process after the second labeled body addition process;
And a procedure for selectively determining one measurement result from a plurality of measurement result candidate values obtained by the first measurement process based on the result of the second measurement process. The analysis method according to claim 1 or 2,
The labeled antibody reagent to said at second labels addition treatment, the label of an amount of 1 / 2-1 / 1000 of the amount of added label in labeled form addition treatment of the first is added The analysis method characterized by adding. The analysis method according to claim 1 or 2,
The second measurement process includes:
A first measurement for measuring a physical quantity related to the label immediately after the second label addition process;
And a second measurement for measuring a physical quantity relating to the label when the predetermined time after the second label addition treatment has elapsed,
One measurement result is selectively determined from a plurality of measurement result candidate values obtained by the first measurement process based on a difference between the first measurement result and the second measurement result. Analysis method to do. The first time when a capture antibody or capture antigen containing a capture body, a labeled antigen or label antibody containing a label, and a first reagent containing a plurality of reaction solid phases are added to a reaction container containing a sample to be analyzed A procedure for performing the addition process of
Second time of adding a second reagent containing a capture antibody or capture antigen containing a capture body, a labeled antigen or labeled antibody containing a label, and a reaction solid phase not contained in the first reagent A procedure for performing the addition process;
Removing at least a part of the reaction solid phase from the reaction vessel, and performing a first measurement process for measuring a physical quantity related to the labeled body including the reaction solid phase to form a bond;
A procedure for performing an addition process of adding a reagent containing a labeled antigen or labeled antibody containing a labeled body to the reaction container after the first measurement process;
A procedure for performing a second reaction solid phase washing treatment after the second labeling agent addition treatment;
A procedure for performing a second measurement process after the second reaction solid phase washing process;
And a procedure for selectively determining one measurement result from a plurality of measurement result candidate values obtained by the first measurement process based on the result of the second measurement process. A sample container for storing a sample to be analyzed;
A labeled reagent container containing a labeled antigen that forms a specific antigen-antibody bond with a component to be analyzed in the sample or a labeled reagent containing a labeled antibody;
A capture reagent container containing a capture antigen that forms a specific antigen-antibody bond with a component to be analyzed in the sample or a capture reagent containing the capture antibody;
A reaction solid phase reagent container that contains a reaction solid phase reagent including a reaction solid phase that forms a specific antigen-antibody bond with a component to be analyzed in the sample;
A reaction container containing a reaction solid phase for dispensing and reacting the sample and the labeling reagent, the capture reagent, and the solid phase reagent;
A sample dispensing mechanism for dispensing the sample from the sample container to the reaction container;
A reagent dispensing mechanism for dispensing the labeling reagent, the supplementary reagent, and the solid phase reagent from the labeling reagent container , the capture reagent container, and the reaction solid phase reagent container , respectively, to the reaction container;
An analysis mechanism for performing qualitative and quantitative analysis of the component to be analyzed based on a physical quantity relating to a label in the reaction container;
A measurement mechanism for performing a measurement process for measuring a physical quantity related to the labeled body in the reaction container;
First capture body addition process for adding the capture reagent to the reaction container containing the sample, first label addition process for adding a labeling reagent to the reaction container, and a reaction solid phase in the reaction container A first solid phase addition treatment for adding a reagent, a first reaction solid phase washing treatment for washing the reaction solid phase in order to remove components other than the components that have formed the antigen-antibody bond including the reaction solid phase, Taking out at least a part of the reaction solid phase from the reaction vessel, and measuring the physical quantity of the labeled body including the reaction solid phase to form a bond, and the measurement process after the first measurement process A second labeling treatment process to the reaction vessel, a second reaction solid phase washing process after the second labeling substance addition process, and a second measurement process after the second reaction solid phase washing process. To the result of the second measurement process Zui, the automatic analyzer characterized by comprising an analysis processor control system for selectively placing one measurement results from a plurality of measurements candidate value obtained by the first measurement process.

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