JP2011158334A - Analyzing method, analyzer, and analyzing program - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide an analyzing method capable of shortening an analyzing treatment time and enhancing analyzing precision, and an analyzer. <P>SOLUTION: In the analyzing method, Raman scattered light Lfw before reaction by gold particles 52 is photometrically measured before the gold particles are bonded to an antigen 50 to be analyzed, and Raman scattered light Lfu after reaction by the gold particles 52 not bonded to the antigen 50 to be analyzed is photometrically measured in a state that the magnetic particles 51 not bonded to the antigen 50 to be analyzed and a composite 53 are removed from laser beam irradiation region after the composite 53 is formed. By operating a difference between the photometrically measured value of the Raman scattered light before reaction and that of the Raman scattered light after reaction, the intensity of the Raman scattered light by the composite 53 can be acquired without measuring the Raman scattered light by the composite 53 itself. <P>COPYRIGHT: (C)2011,JPO&INPIT

Description

  The present invention provides a magnetic reagent containing magnetic particles that specifically bind to an analyte in a sample, and Raman scattering light whose surface is enhanced by irradiation with laser light, and specifically to the analyte. The present invention relates to an analysis method, an analysis apparatus, and an analysis program for analyzing a sample using a labeling reagent containing a labeling substance to be bound.

Raman spectroscopic analysis is an analysis method that detects Raman scattered light generated by irradiating a sample with laser light, and because it can obtain information on the molecular structure of the target, biochemical substances such as viruses and proteins It is known as an effective analysis method for detecting environmental chemicals and biosensors. In this Raman spectroscopic analysis, it is known that the Raman scattering intensity of adsorbed species on the surface of metals such as gold and silver having an atomic level roughness may be enhanced 10 ² -10 ⁶ times compared to non-adsorbed species. This phenomenon is called surface enhanced Raman scattering (SERS).

  As an analysis apparatus using such SERS spectroscopy, a substrate provided with an antibody capable of binding to either the antigen to be analyzed or the surface-enhanced Raman particle is used, and the antibody and the surface-enhanced Raman particle are associated with each other. The difference between the intensity of the Raman scattered light and the intensity of the Raman scattered light when the surface-enhanced Raman particle contacts the antigen to be analyzed and the antigen to be analyzed is associated with the antibody instead of the surface-enhanced Raman scattering particle. An analysis method for detecting an antigen to be analyzed by detection has been proposed (see Patent Document 1). However, in the analysis method described in Patent Document 1, it is necessary to use a substrate having a complicated structure provided with an antibody capable of associating with an antigen to be analyzed for each measurement, and each analysis process is performed on each substrate. There has been a problem that a very complicated analyzer configuration is required to execute them in parallel in the same apparatus.

On the other hand, in recent years, it has been discovered that the Raman scattering intensity can be increased to about 10 ¹⁴ times that of non-adsorbed species by aggregating metal nanoparticles. Therefore, by using metal nanoparticles such as gold and silver having an atomic level roughness as a labeling substance, in addition to this, the complex of the labeling substance and the detection target is aggregated to enhance the Raman scattering intensity. An analysis method for analyzing a detection object has attracted attention. For example, an analyzer that injects a reagent containing magnetic particles that react with a detection target in a sample in addition to a reagent containing metal nanoparticles as a labeling substance has been proposed (see Patent Document 2). ). In this analyzer, a composite of a magnetic particle, a detection target, and a labeling substance is aggregated to a predetermined size by bringing a magnet having a pointed tip from the outside of the reaction container into proximity, and the generated aggregate By irradiating the laser beam to the surface and measuring the surface-enhanced Raman scattered light, the detection target in the specimen is detected without taking a complicated apparatus configuration.

Special table 2007-526488 International Publication No. 2008/014223

  However, in the analyzer described in Patent Document 2, since a magnet with a sharp tip is used to aggregate the complex, the magnet area close to the reaction vessel is reduced, so that the Raman scattered light can be detected. It takes a long time to agglomerate the complex until the entire analysis process takes a long time. Further, in the analyzer described in Patent Document 2, aggregates having a fixed shape are not always formed, but variations occur in the aggregate shape of the complex depending on the dispersion state of the complex. As a result, there is a difference in the irradiation state of the laser beam to the aggregate depending on the aggregate shape, and the intensity of the Raman scattered light measured by the photometer also varies, and it is impossible to perform a high-precision analysis. was there.

  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 analysis apparatus capable of shortening analysis processing time and improving analysis accuracy.

  In order to solve the above-described problems and achieve the object, an analysis method according to the present invention includes a magnetic reagent containing magnetic particles that specifically bind to an analyte in a sample, and laser light irradiation. In the analysis method for analyzing the specimen using a labeling reagent containing a labeling substance that emits surface-enhanced Raman scattered light and specifically binds to the analyte, the specimen, the magnetic reagent, and the labeling reagent A dispensing step in which the magnetic particles and the labeling reagent dispensed in the dispensing step are combined with the analyte in the sample before the magnetic particles are mixed with the laser beam. A pre-reaction photometric step of irradiating the reaction vessel with laser light in a state removed from the irradiation region, and measuring Raman scattered light from the labeling substance in the reaction vessel; A reaction promoting step for promoting a reaction that forms a complex in which the magnetic particles, the analyte and the labeling substance are specifically bound, and the magnetic particles not bound to the analyte after the reaction promoting step and the complex A post-reaction photometric step of irradiating the reaction container with the laser light in a state where the body is removed from the laser light irradiation region, and measuring Raman scattered light by the labeling substance in the reaction container; By calculating the difference or ratio between the photometric value in the photometric step and the photometric value in the post-reaction photometric step, the intensity of the Raman scattered light by the complex is obtained, and the intensity of the Raman scattered light by the acquired complex is obtained. And a calculation step for obtaining the concentration of the analyte in the specimen.

  Further, in the analysis method according to the present invention, in the pre-reaction photometry step, the magnetic particles are removed from the laser beam by generating a magnetic field in a reaction container in which the specimen, the magnetic reagent, and the labeling reagent are dispensed. In the post-reaction photometric step that is removed from the irradiation region, the laser beam is used to convert the magnetic particles that have not been combined with the analyte by generating a magnetic field in the reaction container in which the complex is generated, and the complex. It is characterized by removing from the irradiation area.

  Further, in the analysis method according to the present invention, in the pre-reaction photometry step, a magnet is brought close to an area other than the laser light irradiation area from the outside of the reaction container into which the sample, the magnetic reagent, and the labeling reagent are dispensed. By collecting the magnetic particles, the magnetic particles are collected outside the laser light irradiation region, and the post-reaction photometric step is performed in a region other than the laser light irradiation region from the outside of the reaction vessel in which the complex is generated. By bringing them close to each other, the magnetic particles that have not been combined with the analysis object and the composite are collected outside the irradiation region of the laser beam.

  Moreover, the analysis method according to the present invention is characterized in that the magnet has a size capable of being close to at least a part of a bottom surface or a side surface of the reaction vessel.

  The analysis method according to the present invention is characterized in that there are a plurality of the magnets.

  Further, the analysis method according to the present invention includes the specimen, the magnetic reagent in a reaction container for post-reaction measurement different from the reaction container for pre-reaction measurement in which the Raman scattered light is measured in the pre-reaction photometry step. And a post-reaction measurement dispensing step for dispensing the labeling reagent, wherein the pre-reaction photometric step comprises dispensing a magnetic current collecting rod for generating a magnetic field into the specimen, the magnetic reagent, and the labeling reagent. The magnetic particles are inserted into a reaction vessel for pre-reaction measurement and transferred to the outside of the pre-reaction measurement reaction vessel with the magnetic particles collected by the magnetic rod. In the reaction container for post-reaction measurement in which the sample, the magnetic reagent, and the labeling reagent are dispensed in the post-reaction measurement dispensing step. The post-reaction photometric step promotes the reaction that forms the complex, and the post-reaction photometric step inserts the magnetic collecting rod into the reaction container for post-reaction measurement in which the complex is generated, and binds to the analyte. By transferring the magnetic flux collecting rod out of the reaction vessel for post-reaction measurement in a state where the magnetic particles and the composite that have not been collected on the surface of the magnetic flux collecting rod are not combined with the analyte. The magnetic particles and the composite are removed from the laser light irradiation region.

  Moreover, the analysis method according to the present invention is characterized in that the dispensing step and the pre-reaction photometric step are performed for each specimen.

  The analyzer according to the present invention emits Raman scattered light whose surface is enhanced by irradiation with laser light and a magnetic reagent containing magnetic particles that specifically bind to the analyte in the sample and the analysis. In an analyzer that uses a labeling reagent containing a labeling substance that specifically binds to an object and analyzes the specimen, photometry is performed by measuring the Raman scattered light from the labeling substance by irradiating the reaction container with a laser beam. Removing means for removing the magnetic material from the laser light irradiation region in the reaction container, and calculating means for obtaining the concentration of the analyte in the specimen based on the photometric value obtained by the photometric means, The removing means removes the magnetic particles from the irradiation region of the laser beam before the magnetic particles and the labeling reagent are combined with the analyte in the specimen. And a post-reaction removal process for removing the magnetic particles that have not been combined with the analyte after the complex is generated by the reaction means and the complex from the laser light irradiation region, and the photometry The treatment is performed by irradiating the laser beam into the reaction container in a state where the magnetic particles are removed from the laser light irradiation region by the pre-reaction removal process by the removing unit, and using the labeling substance in the reaction container. A state in which the magnetic particles that have not been combined with the analyte and the complex are removed from the irradiation region of the laser light by the pre-reaction photometric process for measuring Raman scattered light and the post-reaction removal process by the removing unit And performing post-reaction photometric processing in which the laser light is irradiated into the reaction container to measure Raman scattered light from the labeling substance in the reaction container, and the computing means includes: The intensity of Raman scattered light by the complex is obtained by calculating the difference or ratio between the photometric value in the pre-reaction photometric process and the post-reaction photometric process by the photometric means, and the obtained complex The concentration of the analyte in the specimen is obtained based on the intensity of the Raman scattered light generated by.

  In the analyzer according to the present invention, the removing means generates the magnetic particles by generating a magnetic field in a reaction container in which the sample, the magnetic reagent, and the labeling reagent are dispensed in the pre-reaction removal process. The magnetic particles that have not been combined with the analyte by being removed from the laser light irradiation region and generating a magnetic field in the reaction container in which the complex is generated in the post-reaction removal process, and the complex It removes from the irradiation area of the said laser beam, It is characterized by the above-mentioned.

  An analysis program according to the present invention causes an analysis apparatus to execute any one of the analysis methods described above.

  The present invention measures the Raman scattered light before the reaction with the labeling substance before binding to the antigen or antibody to be analyzed, and magnetic particles that have not bound to the complex and the antigen or antibody to be analyzed after complex formation. Measure the Raman scattered light after the reaction with the labeling substance that did not bind to the antigen or antibody to be analyzed in the state where is removed from the irradiation area of the laser light, the photometric value of the Raman scattered light before the reaction and the Raman scattering after the reaction By calculating the difference or ratio with the photometric value of light, the intensity of the Raman scattered light from the complex can be obtained without measuring the Raman scattered light from the complex itself. There is no need to form it, and analysis processing time can be shortened and analysis accuracy can be improved.

FIG. 1 is a schematic diagram illustrating the configuration of the analyzer according to the first embodiment. FIG. 2 is a diagram showing the configuration of the photometry unit shown in FIG. FIG. 3 is a flowchart showing a processing procedure of analysis processing by the analysis apparatus shown in FIG. FIG. 4 is a diagram for explaining analysis processing by the analyzer shown in FIG. FIG. 5 is a diagram showing the relationship between the reaction time after the stirring process and the number of unreacted particles in the analyzer shown in FIG. FIG. 6 is a flowchart showing a processing procedure of the arithmetic processing shown in FIG. FIG. 7 is a Raman spectrum obtained by the analysis apparatus shown in FIG. FIG. 8 is a diagram for explaining a conventional analysis process. FIG. 9 is a diagram showing another configuration of the photometry unit shown in FIG. FIG. 10 is a flowchart showing another processing procedure of the arithmetic processing shown in FIG. FIG. 11 is a schematic diagram illustrating the configuration of the analyzer according to the second embodiment. FIG. 12 is a perspective view schematically showing the magnetic flux collecting rod transfer unit and the magnetic flux collecting rod cleaning unit shown in FIG. FIG. 13 is a flowchart showing a processing procedure of analysis processing by the analysis apparatus shown in FIG. FIG. 14 is a flowchart showing a processing procedure of the pre-reaction measurement process shown in FIG. FIG. 15 is a diagram for explaining the pre-reaction measurement process shown in FIG. FIG. 16 is a flowchart showing a processing procedure of the post-reaction measurement process shown in FIG. FIG. 17 is a diagram for explaining the post-reaction measurement process shown in FIG. FIG. 18 is a flowchart illustrating a processing procedure of the arithmetic processing illustrated in FIG. FIG. 19 is a flowchart showing another processing procedure of the arithmetic processing shown in FIG.

  Hereinafter, an analyzer that is an embodiment according to the present invention will be described using an analyzer that performs analysis on a body fluid sample such as blood, urine, or saliva using SERS spectroscopy. Note that the present invention is not limited to the embodiments. In the description of the drawings, the same parts are denoted by the same reference numerals.

(Embodiment 1)
First, the first embodiment will be described. FIG. 1 is a schematic diagram illustrating the configuration of the analyzer according to the first embodiment. As shown in FIG. 1, the analyzer 1 according to the first embodiment emits laser light into a reaction vessel containing a labeling substance, and measures a surface-enhanced Raman scattered light by the labeling substance. And a control mechanism 4 that controls the entire analyzer 1 including the measurement mechanism 2 and that analyzes the measurement results in the measurement mechanism 2. The analyzer 1 automatically performs analysis on a plurality of specimens by the cooperation of these two mechanisms.

  The measurement mechanism 2 will be described. The measurement mechanism 2 is roughly divided into a reaction table 10, a sample transfer unit 19, a sample dispensing unit 20, a magnetic reagent container 21, a magnetic reagent dispensing unit 22, a probe washing unit 23, a labeling reagent container 24, and a labeling reagent dispensing unit. 25, a probe cleaning unit 26, a stirring unit 27, and a photometry unit 30 are provided.

  The reaction table 10 transports the reaction container 11 to a predetermined position in order to perform dispensing of a sample or a reagent to the reaction container 11, stirring the reaction container 11, and photometry while holding the reaction container 11 at a constant temperature. The reaction table 10 is rotatable about a vertical line passing through the center of the reaction table 10 as a rotation axis by driving a drive mechanism (not shown) under the control of the control unit 41 described later.

  The sample transfer unit 19 includes a plurality of sample racks 19b that hold a plurality of sample containers 19a that store liquid samples such as blood, urine, or saliva and sequentially transfer them in the direction of the arrows in the figure. The sample in the sample container 19a transferred to the sample aspiration position on the sample transfer unit 19 is dispensed by the sample dispensing unit 20 into the reaction container 11 that is arranged and transported on the reaction table 10.

  The sample dispensing unit 20 includes an arm that freely moves up and down in the vertical direction and rotates around a vertical line passing through its base end as a central axis. A sample probe for aspirating and discharging the sample is attached to the tip of the arm. The sample dispensing unit 20 includes an intake / exhaust mechanism using an unillustrated intake / exhaust syringe or piezoelectric element. The sample dispensing unit 20 sucks the sample from the sample container 19a transferred to the predetermined position on the sample transfer unit 19 with the sample probe, rotates the arm counterclockwise in the drawing, The specimen is discharged into the reaction container 11.

  The magnetic reagent storage 21 can store a plurality of magnetic reagent bottles 21a in which magnetic reagents are stored. The labeling reagent storage 24 can store a plurality of labeling reagent bottles 24a in which labeling reagents are stored. The magnetic reagent in the magnetic reagent bottle 21a and the labeling reagent in the labeling reagent bottle 24a are respectively dispensed into the reaction container 11 of the reaction table 10. The magnetic reagent container 21 and the labeled reagent container 24 can be rotated clockwise or counterclockwise by driving a drive mechanism (not shown), and a desired reagent bottle can be dispensed with the magnetic reagent dispensing unit 22 or the labeled reagent dispensing. Transfer to the reagent suction position by the unit 25. Here, the analyte in the sample includes, for example, antibodies, proteins, peptides, amino acids, carbohydrates, hormones, steroids, vitamins, bacteria, DNA, RNA, cells, viruses, any antigenic substances, haptens, antibodies and There are combinations of these. A magnetic reagent is a reagent containing magnetic particles solid-phased with an antigen or antibody that binds to the analyte in the sample that is the subject of analysis. When the magnetic particles are located in a region to which the magnetic field extends, the magnetic particles have a magnetic force in the magnetic field and are attracted to the magnetic field generation source. The labeling reagent is a reagent containing a labeling substance that binds to the analyte in the sample. The labeling substance is a nanoparticle containing a metal such as gold, silver, copper, or platinum having an atomic level surface roughness, and the surface of the particle is coated with an antigen or an antibody that can bind to an analyte. ing. Even if this labeling substance is in the form of a labeling substance alone or a complex bound to the analyte, it emits Raman-enhanced light with enhanced surface when irradiated with laser light. In the following description, a case where gold particles are used as the labeling substance will be described as an example.

  The magnetic reagent dispensing unit 22 has a reagent probe for aspirating and discharging the magnetic reagent attached to the distal end, and freely moves up and down in the vertical direction and rotates around the vertical line passing through its base end as a central axis. Provide an arm. The magnetic reagent dispensing unit 22 includes an intake / exhaust mechanism using an unillustrated intake / exhaust syringe or piezoelectric element. The magnetic reagent dispensing unit 22 sucks the reagent in the magnetic reagent bottle 21a moved to a predetermined position by the magnetic reagent container 21 by the reagent probe, rotates the arm, and is transported to the magnetic reagent discharge position by the reaction table 10. Dispense into the reaction vessel 11. The labeling reagent dispensing unit 25 has the same configuration as the magnetic reagent dispensing unit 22, and the reagent in the labeled reagent bottle 24a moved to a predetermined position by the labeling reagent storage 24 is aspirated by the reagent probe, and the arm is moved. It is swung and dispensed into the reaction vessel 11 conveyed to the labeling reagent discharge position by the reaction table 10.

  The probe cleaning units 23 and 26 are arranged on the locus of the reagent probe of the magnetic reagent dispensing unit 22 and on the locus of the reagent probe of the labeled reagent dispensing unit 25, respectively, and clean the inside and outside of the reagent probe. After the reagent dispensing is completed, the reagent probe is washed by the probe washing units 23 and 26, and then the next reagent dispensing process is performed.

  The stirring unit 27 stirs the sample dispensed into the reaction container 11 and the reagent to promote the reaction. The sample dispensed in the reaction vessel 11, the magnetic particles contained in the magnetic reagent and the labeling reagent, the antibody or antigen to be analyzed, and the labeling substance are stirred by the stirring unit 27 to promote the reaction, and a predetermined reaction As time elapses, the analyte contained in the specimen, the magnetic particles, and the labeling substance are combined to form a complex. This composite has magnetic particles. Further, since this complex has a labeling substance, when the laser beam is irradiated, the surface-enhanced Raman scattered light is emitted by the labeling substance.

  The photometry unit 30 irradiates the reaction vessel 11 with laser light, and measures the surface-enhanced Raman scattering light by the labeling substance in the reaction vessel 11. For example, as shown in FIG. 2, the photometry unit 30 includes a laser light source 30a, a Raman spectrometer 30b, and lenses 30d, 30e, 30f, and 30g, and has a configuration in which laser light is incident on the side surface of the reaction vessel 11 from an oblique direction. . The laser light Li emitted from the laser light source 30a is collimated by the lens 30d and then condensed by the lens 30e and enters the reaction container 11 from an oblique direction with respect to the side surface of the reaction container 11. This laser beam Li irradiates the gold particles 52 located in the irradiation region S1 in the reaction vessel 11. As a result, the Raman scattered light Lf enhanced on the surface of the gold particle 52 is aligned with the parallel light by the lens 30f, collected by the lens 30g, and then incident on the Raman spectrometer 30b. The measurement result of the Raman spectrometer 30b is output to the control unit 41, and the analysis unit 43 analyzes the sample based on the measurement result.

  Then, as shown in FIG. 2, the photometry unit 30 further includes a permanent magnet 30c provided outside the reaction vessel 11 so as to be close to a region other than the irradiation region S1 of the laser light Li from the laser light source 30a. Have. The permanent magnet 30 c is disposed at a position close to the bottom surface of the reaction vessel 11 when the reaction vessel 11 is transferred to the upper portion by the reaction table 10. The permanent magnet 30 c has a size that can approach almost the entire bottom surface of the reaction vessel 11. The permanent magnet 30c collects the magnetic particles in the reaction vessel 11 and the magnetic particles constituting the composite on the bottom surface side of the reaction vessel 11 by generating a magnetic field inside the reaction vessel 11 located above the permanent magnet 30c. . In other words, the permanent magnet 30c removes magnetic particles and composites in the reaction vessel 11 from the irradiation region S1 of the laser beam Li by generating a magnetic field inside the reaction vessel 11 located above the permanent magnet 30c. . The permanent magnet 30c is provided only at the photometry position by the photometry unit 30, and the reaction vessel 11 transferred to the photometry position in the photometry unit 30 by the transfer process of the reaction table 10 is attached to the bottom surface of the reaction vessel 11 with the permanent magnet 30c. Are positioned so as to be close to each other, the internal magnetic particles and the composite are removed from the irradiation region S1 of the laser beam Li. In the reaction vessel 11 transferred to a position other than the photometric position by the reaction table 11, the magnetic particles and the composite are dispersed in the liquid in the reaction vessel 11 because they are not collected by the permanent magnet 30 c. Yes.

  Note that the reaction vessel 11 for which the analysis process has been completed is discarded by a discard mechanism (not shown) for each analysis process. Further, the analyzer 1 may be provided with a container cleaning mechanism (not shown) to clean the reaction container 11 after the analysis process and then use it again for the analysis process. In this case, the container cleaning mechanism sucks and discharges the mixed liquid in the reaction container 11 with a nozzle, and cleans the reaction container 11 by injecting and sucking a cleaning liquid such as a detergent or cleaning water.

  Next, the control mechanism 4 will be described. The control mechanism 4 includes a control unit 41, an input unit 42, an analysis unit 43, a storage unit 44, and an output unit 45. These units included in the measurement mechanism 2 and the control mechanism 4 are electrically connected to the control unit 41.

  The control unit 41 is configured using a CPU or the like, and controls processing and operation of each unit of the analyzer 1. The control unit 41 performs predetermined input / output control on information input / output to / from each of these components, and performs predetermined information processing on this information. The input unit 42 is configured using a keyboard, a mouse, and the like, and acquires various information necessary for analyzing the sample, instruction information for the analysis operation, and the like from the outside. The analysis unit 43 analyzes the sample based on the Raman spectrophotometric value acquired from the photometry unit 30. The storage unit 44 is configured by using a hard disk that magnetically stores information and a memory that electrically loads various programs related to the process from the hard disk when the analyzer 1 executes the process, Various information including the analysis result of the sample is stored. The storage unit 44 may include an auxiliary storage device that can read information from a storage medium such as a CD-ROM, a DVD-ROM, or a PC card. The output unit 45 is configured using a printer, a speaker, and the like, and outputs various information including the analysis result of the sample. The output unit 45 may output information according to a predetermined format to an external device via a communication network (not shown).

  In the analyzer 1 having the above configuration, the intensity of scattered light by the complex itself in which the analyte, the magnetic particles, and the labeling substance contained in the sample are combined is not measured. The analyzer 1 measures the intensity of Raman scattered light that corresponds to the total amount of labeled substance dispensed before the reaction before binding to the analyte, and further generates a complex by a reaction promoting process. After that, the intensity of the Raman scattered light of the labeling substance not bound to the analysis object is measured in a state where the magnetic particles and the complex are removed. And in the analyzer 1, the difference of the intensity | strength of the Raman scattered light corresponded to the whole quantity of the labeled | labeled substance before reaction dispensed, and the Raman scattered light by the labeled substance after the reaction which was not couple | bonded with the analyte. By obtaining the value, the intensity of the Raman scattered light of the complex itself is indirectly acquired, and the concentration of the analyte in the sample is detected based on this difference value.

  Therefore, with reference to FIG. 3 and FIG. 4, the analysis processing of the analyzer 1 will be described in detail. FIG. 3 is a flowchart showing a processing procedure of analysis processing by the analysis apparatus 1 shown in FIG. 1, and FIG. 4 is a diagram for explaining analysis processing by the analysis apparatus 1 shown in FIG.

  In the analyzer 1, when the analysis process start is instructed, as shown in FIGS. 3 and 4 (1), the magnetic reagent dispensing unit 22 is transferred to a predetermined position by the magnetic reagent container 21. A magnetic reagent dispensing process is performed in which the magnetic reagent containing the magnetic particles 51 is dispensed into the reaction container 11 located at the magnetic reagent dispensing position of the reaction table 10 from within 21a (step S1). Next, as shown in FIG. 4B, the sample dispensing unit 20 dispenses the sample to be analyzed from the sample container 19a into the reaction container 11 into which the magnetic reagent has been dispensed (step S2). . The specimen includes the antigen 50 that is the analysis target, and also contains a contaminant 55 other than the antigen 50. In the specimen dispensing process, the contaminant 55 is also contained in the reaction container 11 together with the antigen 50. Dispensed within. Then, as shown in FIG. 4 (3), the labeling reagent dispensing unit 25 is capable of binding to the antigen 50 to be analyzed from the labeling reagent bottle 24 a moved to a predetermined position by the labeling reagent storage 24. A labeling reagent dispensing process is performed in which the labeling reagent containing 52 is dispensed into the reaction container 11 positioned at the labeling reagent dispensing position of the reaction table 10 and into which the magnetic reagent and the sample have been dispensed ( Step S3). Thereafter, the reaction table 10 transfers the reaction vessel 11 into which the magnetic reagent, the specimen, and the labeling reagent have been dispensed to the agitation position by the agitation unit 27, and the agitation unit 27 agitates the liquid in the reaction vessel 11. (Step S4).

  After completion of the stirring process by the stirring unit 27, the reaction table 10 transfers the reaction container 11 to the photometric position by the photometric unit 30 before the binding reaction of the magnetic particles 51, the antigen 50, and the gold particles 52 proceeds. In the photometry unit 30, as described with reference to FIG. 2, the permanent magnet 30c is close to the bottom surface of the reaction vessel 11 to be photometric.

  As a result, as shown in FIG. 4 (4), the antigen 50 in the reaction vessel 11 and the magnetic particles 51 before binding are collected on the bottom surface of the reaction vessel 11 close to the permanent magnet 30c. In other words, the magnetic particles 51 before being bound to the antigen 50 are removed from the irradiation region of the laser light Li. In the photometry unit 30, in this state, the laser light source 30a irradiates the reaction container 11 with the laser light Li, and the Raman spectrometer 30b determines the intensity of the Raman scattered light Lfw by the gold particles 52 before binding to the antigen 50. A pre-reaction photometry process (step S5) for photometry is performed. Here, since the magnetic particle 51 has a diameter that is 10 times or more that of the gold particle 52 that is the labeling substance and also has a property of absorbing the laser light Li, the laser light reaches the gold particle 52 by the magnetic particle 51. May be inhibited. In the pre-reaction photometric process, by collecting the magnetic particles 51 outside the irradiation region of the laser light Li, only the gold particles 52, the antigen 50 in the specimen, and the contaminants 55 are within the irradiation region in the reaction vessel 11. In the existing state, the Raman scattered light corresponding to the total amount of the dispensed gold particles 52 is accurately measured. Since the magnetic particles 51 are collected by bringing the permanent magnet 30c close to almost the entire bottom surface of the reaction vessel 11, the time required for collecting the light by the photometry unit 30 is about 10 seconds. Further, as shown in FIG. 5 showing the relationship between the reaction time after the stirring process and the number of unreacted particles, the pre-reaction photometric process is executed whenever it is during a period Tw before the number of unreacted particles starts to decrease. For example, it is executed at the timing when the reaction time has elapsed 5 minutes. In FIG. 4 (4), the light condensing mechanism in the photometry unit 30 is not shown.

  And after this pre-reaction photometric process is complete | finished, the stirring part 27 performs a stirring process again with respect to this reaction container 11 (step S6), and accelerates | stimulates reaction. Then, the analyzer 1 performs a reaction promoting process for promoting an antigen-antibody reaction that specifically binds the magnetic particles 51, the antigen 50, and the gold particles 52 in the reaction container for a predetermined time (step S7), and FIG. As shown in 5), a complex 53 in which the magnetic particles 51, the antigens 50, and the gold particles 52 are specifically bound is generated in the reaction vessel 11. As shown in FIG. 5, it is sufficient that the reaction time is the time included in the period Tu in which the number of unreacted particles that have decreased is constant and the reaction with the antigen is considered to be almost completed.

  Next, the reaction table 10 transfers the reaction container 11 in which the complex has been generated after a predetermined reaction time to the photometric position by the photometric unit 30. In the photometry unit 30, as in the pre-reaction photometry process (step S3), as shown in FIG. 4 (6), the permanent magnet 30c is in close proximity to the bottom surface of the reaction vessel 11 to be photometrically measured. Magnetic particles 51 are collected on the bottom surface of the reaction vessel 11. In the reaction container 11 after the complex 53 is formed, magnetic particles constituting the complex 53 are mixed as a magnetic substance in addition to the magnetic particles 51 that are not bound to the antigen 50 to be analyzed. For this reason, in the reaction container 11 after the complex 53 is formed, the magnetic particles 51 and the magnetic particles of the complex 53 are collected on the bottom surface of the reaction container 11 by the permanent magnet 30c. Along with this, the gold particles that bind to the antigen 50 and form the complex 53 are also collected on the bottom surface side of the reaction vessel 11 and removed from the irradiation region of the laser beam Li.

  In this state, the laser light source 30a irradiates the reaction vessel 11 with the laser light Li, and the Raman spectrometer 30b is subjected to Raman scattered light by the gold particles 52 that are not bound to the antigen 50 that is the analysis target. A post-reaction photometric process (step S8) for measuring the intensity of Lfu is performed. In this post-reaction photometric processing, the magnetic particles 51 and the magnetic particles constituting the complex 53 are collected outside the laser light Li irradiation region, so that they do not bind to the antigen 50 in the irradiation region in the reaction vessel 11. In the state where only the gold particles 52 and the impurities 55 in the specimen are present, the Raman scattered light corresponding to the gold particles 52 not bound to the antigen 50 is accurately measured. In the analyzer 1, the permanent magnet 30c is brought close to almost the entire bottom surface of the reaction vessel 11 to collect the magnetic particles 51 and the magnetic particles of the composite 53, so that efficient magnetic collection processing is possible and photometry is performed. The magnetism collecting time by the unit 30 is sufficient for a short time of about 10 seconds. In addition, as shown in FIG. 5 showing the relationship between the reaction time after the stirring treatment and the number of unreacted particles, the post-reaction photometric treatment has a constant reduced number of unreacted particles, and the reaction with the antigen is almost completed. It may be executed at any time as long as it is during the period Tu considered to have been performed, for example, at a timing when a reaction time of 35 minutes has elapsed. In FIG. 4 (6), the light condensing mechanism in the photometric unit 30 is not shown.

  Then, the analysis unit 43 performs a calculation process for obtaining the concentration of the antigen 50 to be analyzed in the sample based on the photometric value in the pre-reaction photometric process (step S3) and the photometric value in the post-reaction photometric process (step S9). ). Thereafter, the output unit 45 performs an output process for outputting the concentration of the antigen 50 obtained by the analysis unit 43 (step S10), and ends the analysis process in the analyzer 1. The control unit 41 may store the concentration of the antigen 50 obtained by the analysis unit 43 in the storage unit 44.

  Next, the arithmetic processing shown in FIG. 3 will be described with reference to FIG. FIG. 6 is a flowchart showing a processing procedure of the arithmetic processing shown in FIG. As shown in FIG. 6, the analysis unit 43 acquires a pre-reaction photometric value (step S20), and acquires a post-reaction photometric value (step S22). Then, the analysis unit 43 calculates a difference value between the acquired pre-reaction photometric value and the post-reaction photometric value (step S24).

  Specifically, by measuring the Raman scattered light with the Raman spectrometer 30b, a Raman spectrum having a Raman scattering intensity on the vertical axis and a Raman shift on the horizontal axis as shown in FIG. 7 can be obtained as a measurement result. . Here, the curve Lw shown in FIG. 7 is a Raman spectrum obtained in the pre-reaction photometry process, and the curve Lu is a Raman spectrum obtained in the post-reaction photometry process. The analysis unit 43 acquires a value Iw obtained by subtracting the base value from the maximum intensity value in the curve Lw as a pre-reaction photometric value, and acquires a value Iu obtained by subtracting the base value from the maximum intensity value in the curve Lu as a pre-reaction photometric value. .

  This pre-reaction photometric value corresponds to the intensity of Raman scattered light corresponding to the total amount of gold particles 52 dispensed in the reaction vessel 11 as described above. As described above, the post-reaction photometric value corresponds to the intensity of Raman scattered light corresponding to the gold particle 52 that has not bound to the antigen 50 because the complex 53 is removed. For this reason, the difference value between the pre-reaction photometric value and the post-reaction photometric value is determined by the Raman scattering by the gold particles constituting the complex 53 by combining with the antigen 50 removed from the irradiation region of the laser light Li in the post-reaction photometric process. Corresponds to the intensity of light. That is, the difference value between the pre-reaction photometric value and the post-reaction photometric value corresponds to the intensity of Raman scattered light by the complex 53. In the analyzer 1, the analysis unit 43 calculates the difference value between the pre-reaction photometric value and the post-reaction photometric value, thereby indirectly acquiring the intensity of Raman scattered light from the complex 53.

  Then, the analysis unit 43 calculates an analysis value corresponding to the difference value based on the difference value corresponding to the intensity of the Raman scattered light by the complex 53 (step S26), and the control unit 41 calculates the calculation result. Is output, and the arithmetic processing ends.

  Thus, in the analyzer 1, the analyzer 1 measures the intensity of Raman scattered light by the labeling substance before the reaction before binding to the analyte, and after the complex is generated by the reaction promoting process, Measure the Raman scattered light intensity of the labeling substance that has not been bound to the analyte with the body removed, and bind to the analyte after the reaction and the intensity of the Raman scattered light from the labeling substance before the reaction. The concentration of the analyte in the sample is detected by indirectly obtaining the intensity of the Raman scattered light of the complex itself by obtaining the difference value from the Raman scattered light by the labeling substance that has not been present.

  Here, a conventional analysis process will be described. Conventionally, as shown in FIGS. 8 (1) to 8 (3), after dispensing the magnetic particles 51, the specimen containing the antigen 50 and the contaminants 55, and the gold particles 52, FIG. 8 (4) As shown, these are reacted to form a complex 53. Next, conventionally, as shown in FIG. 8 (5), a permanent magnet 130 c having a sharp tip is brought close to the bottom surface of the reaction container 11 to form an aggregate 54 on the bottom surface of the reaction container 11. Conventionally, as shown in FIG. 8 (6), the aggregate 54 is irradiated with the laser light Li 0 from the laser light source 130 a, and the Raman spectrometer 130 b emits the surface-enhanced Raman scattered light Lf 0 from the aggregate 54. I was measuring.

  In such a conventional analysis process, an aggregate having a fixed shape is not always formed, and the shape and size of the aggregate 54 vary depending on the dispersion state of the complex 53 and the like. As a result of the variation in the shape of the aggregate 54, a difference also occurs in the irradiation state of the laser beam Li0 to the aggregate 54. Therefore, the intensity of the Raman scattered light Lf0 measured by the Raman spectrometer 130b also varies, resulting in high accuracy. There was a problem that the analysis of could not be performed.

  Actually, the measurement results of Raman scattered light from the aggregate 54 formed in accordance with the conventional analysis process are shown in Table 1. Table 1 is a table showing the measured values of each Raman scattered light and the coefficient of variation CV (Coefficient of Variation) value when this conventional analysis process is performed 10 times under the same conditions. The used specimen contains TSH antigen at a concentration of 200 [μIU / ml]. The variation coefficient CV value is a value obtained by dividing the standard deviation by the average by a ratio of 100 minutes, and indicates a relative error with respect to the average value.

  As shown in Table 1, when the aggregate 54 is formed according to the conventional analysis process, the CV value of each Raman scattered light is a very high value of 4.3%. As shown in Table 1, when the conventional analysis process is used, the intensity of the Raman scattered light obtained by the photometric process includes a large variation, and therefore the analysis calculated based on the photometric value The results also included variations.

  On the other hand, the analyzer 1 according to the first embodiment does not measure the Raman scattered light from the aggregate 54 in which the aggregate shape varies as in the conventional case, that is, the Raman scattered light from the complex 53 itself. . In the analyzer 1, the intensity of Raman scattered light corresponding to the total amount of the gold particles 52 before binding to the analysis object is measured, and after the complex is generated by the reaction promoting process and the complex is removed. The intensity of the Raman scattered light of the gold particles 52 not bonded to the analysis object is measured. Then, in any photometric process, the analyzer 1 removes the magnetic particles 51 having a diameter 10 times or more that of the gold particles 52 and also has the property of absorbing the laser light Li, and measures the impurities derived from the specimen. In the state where only the target gold particles 52 are dispersed, the Raman scattered light by the gold particles 52 to be measured is measured. In other words, the analyzer 1 does not combine the Raman scattered light corresponding to the total amount of the dispensed gold particles 52 and the analysis object in a state where the arrival of the laser light to the gold particles 52 is not hindered. Since both the Raman scattered light of the gold particles 52 can be measured, the intensity of the Raman scattered light corresponding to the total amount of the dispensed gold particles 52 and the gold particles 52 not bound to the analysis object Accurate values can be obtained for both the intensity of the Raman scattered light.

  And the analyzer 1 is a difference between the intensity of the Raman scattered light corresponding to the total amount of the gold particles 52 before the reaction and the intensity of the Raman scattered light by the labeling substance that has not been bonded to the analysis target after the reaction. By obtaining the value, the intensity of Raman scattered light of the complex is indirectly acquired, and the concentration of the analyte in the sample is detected based on this difference value. Therefore, the analysis apparatus 1 uses either the intensity of the Raman scattered light corresponding to the total amount of the gold particles 52 before the reaction or the intensity of the Raman scattered light by the labeling substance that has not been bonded to the analysis target after the reaction. Therefore, by using the pre-reaction photometric value and the post-reaction photometric value, it is possible to obtain an accurate value of the intensity of the Raman scattered light of the complex itself.

  Table 2 shows the results of the pre-reaction photometry process and the post-reaction photometry process when the analysis process shown in FIGS. Table 2 shows the pre-reaction photometric values in the pre-reaction photometric process and the reaction in the post-reaction photometric process when the analysis process is performed 10 times in the analyzer 1 under the same conditions for the same sample as in Table 1. It is a table | surface which shows the post-photometry value, the difference value of the photometry value before reaction, and the photometry value after reaction, and the variation coefficient CV value of each value.

  As shown in Table 2, in the analyzer 1, the CV value of the pre-reaction photometric value is 0.65%, and the CV value of the post-reaction photometric value is 1.30%, both of which are compared with the conventional one. Indicates a low value. This is because the pre-reaction photometry process and the post-reaction photometry process are performed in a state in which the magnetic particles 51 and the magnetic particles of the complex 53 are excluded from the irradiation region of the laser beam Li. In a state where the arrival is not hindered, the Raman scattered light corresponding to the total amount of the gold particles 52 before the reaction and the Raman scattered light by the labeling substance that has not been bound to the analyte after the reaction are accurately obtained. This is because photometry is possible.

  As shown in Table 2, the CV value of the difference value between the pre-reaction photometric value and the post-reaction photometric value is as low as 0.59%. This is because both the pre-reaction photometric value and the post-reaction photometric value have a small variation, and the difference between these values is also small. Since this difference value corresponds to the intensity of the Raman scattered light of the complex 53 itself, the analyzer 1 calculates the intensity of the Raman scattered light of the complex 53 itself with very little variation by calculating this difference value. Can be obtained stably. Furthermore, in the analyzer 1, since the post-reaction photometric process is performed on the same reaction vessel as the reaction vessel 11 that has performed the pre-reaction photometric process, it is not necessary to consider the photometric error due to the reaction container difference. Therefore, the analysis apparatus 1 can significantly improve the analysis accuracy as compared with the prior art.

  Further, in the conventional analysis method, as shown in FIG. 8 (5), since the permanent magnet 130c having a sharp tip is used for aggregating the composite 53, the magnet area close to the reaction vessel is reduced. A long time is required to agglomerate the complex to such a size that the scattered light can be detected, and there is a problem that the processing time of the entire analysis process becomes long. Specifically, in the conventional analysis process, about 40 minutes are required as the reaction time for forming the complex 53, and the processing time of about 2 minutes is required for the magnetic flux collecting process for forming the aggregate 54. Was necessary. Therefore, in the conventional analyzer, there is a limit in increasing the number of sample processes per unit time, and the processing capacity of the analysis process cannot be improved.

  On the other hand, since the analyzer 1 does not measure the Raman scattered light of the complex 53 itself, in the pre-reaction photometric process and the post-reaction photometric process, the magnetic particles 51, the complex It is sufficient that the magnetic particles of the body 53 can be collected on the bottom surface side of the reaction vessel 11. For this reason, in the analyzer 1, as shown in FIGS. 4 (5) and 4 (6), it becomes possible to use a large permanent magnet 30c that can be brought close to almost the entire bottom surface of the reaction vessel 11. The inner magnetic particles 51 and the magnetic particles 51 of the composite 53 can be efficiently collected on the bottom surface of the reaction vessel 11. Actually, the magnetic collection time in the pre-reaction photometry process and the post-reaction photometry process performed by the analyzer 1 is sufficient as short as about 10 seconds as described above. Therefore, according to the analyzer 1 according to the first embodiment, the processing time of the entire analysis process can be shortened more reliably than before, and the processing capacity of the analysis process can be increased by increasing the number of sample processes per unit time. Can be improved. Of course, as shown in FIG. 1, the analysis apparatus 1 can execute a plurality of analysis processes on a plurality of reaction vessels in parallel in the same apparatus without taking a complicated apparatus configuration. Each process can proceed promptly without stagnation.

  In the first embodiment, the photometric unit 30 having the configuration shown in FIG. 2 has been described as an example. However, the configuration is not limited to that shown in FIG. For example, if the permanent magnet 30c can collect the magnetic particles in the reaction container 11 and the magnetic particles constituting the composite on the bottom surface side of the reaction container 11, the permanent magnet 30c is not the entire bottom surface of the reaction container 11 but the entire surface of the reaction container 11. It may be close to at least a part of the bottom surface. For example, as shown in FIG. 9, the photometry unit 30 may have a configuration in which laser light is incident on the bottom surface of the reaction vessel 11 from an oblique direction. In this case, the laser light Li emitted from the laser light source 30a below the reaction vessel 11 is converged to parallel light by the lens 30d, and then collected by the lens 30e, and is incident on the bottom surface of the reaction vessel 11 from an oblique direction. To do. This laser beam Li irradiates the gold particles 52 located in the irradiation region S11 in the reaction vessel 11. As a result, the Raman scattered light Lf enhanced on the surface of the gold particle 52 is converged to parallel light by the lens 30f, collected by the lens 30g, and then incident on the Raman spectrometer 30b.

  Further, the permanent magnet 30c of the photometry unit 30 is positioned outside the side surface of the reaction vessel 11 as shown in FIG. 9 as long as it is close to a region other than the irradiation region S11 of the laser light Li from the laser light source 30a. You may do it. Further, the permanent magnet 30c is not limited to a single magnet, and a plurality of permanent magnets 30c may be provided as shown in FIG. Furthermore, instead of the permanent magnet 30c, the photometry unit 30 may use an electromagnet formed by a coil wound with a conducting wire formed of a magnetic material as a magnetic field generating member. In this case, the control unit 41 controls the generation and stop of the magnetic field by the electromagnet by controlling the current supplied to the electromagnet, and the current is measured at least while the reaction container to be measured is located at the photometry position of the photometry unit 30. To generate a magnetic field from the electromagnet.

  In Embodiment 1, the analysis unit 43 obtains the Raman scattered light intensity of the labeling substance of the complex by calculating the difference value between the pre-reaction photometric value and the post-reaction photometric value. By calculating the ratio between the photometric value and the post-reaction photometric value, the relative intensity of the Raman scattered light of the complex labeling substance may be obtained to detect the analyte concentration in the sample.

  In this case, as shown in the flowchart of FIG. 10, the analysis unit 43 acquires the pre-reaction photometric value (step S120) and acquires the post-reaction photometric value, similarly to steps S20 to S23 shown in FIG. Step S122). Then, the analysis unit 43 calculates a ratio between the pre-reaction photometric value and the post-reaction photometric value (step S124), calculates an analysis value for the calculated ratio (step S126), and sends the calculation result to the control unit 41. Output, and the arithmetic processing ends. This calculated ratio corresponds to the relative intensity of the Raman scattered light of the labeling substance of the complex with respect to the concentration of the labeling reagent dispensed in the reaction container 11, and the analysis unit 43 is dispensed in the reaction container 11. Based on the concentration of the labeling reagent and the calculated ratio value, the concentration of the antigen or antibody to be analyzed in the sample is calculated.

(Embodiment 2)
Next, a second embodiment will be described. In the analyzer according to the second embodiment, the magnetic particles and the complex in the reaction vessel are removed from the reaction vessel 11 by using a magnetic collecting rod that can be directly inserted into the reaction vessel, so that the magnetic particles, The composite is removed from the laser light irradiation area.

  FIG. 11 is a diagram illustrating a configuration of the analyzer according to the second embodiment. As shown in FIG. 11, in the analysis apparatus 201 according to the second embodiment, the measurement mechanism 202 has a reaction table 210 and a photometry unit 30 instead of the reaction table 10 as compared with the analysis apparatus 1 shown in FIG. Instead of this, a photometric unit 230 is provided, and a magnetic flux collecting rod transfer unit 228 for transferring a magnetic flux collecting rod capable of collecting magnetic material and a magnetic flux collecting rod cleaning unit 229 for cleaning the magnetic flux collecting rod are further provided. The control mechanism 204 of the analysis apparatus 201 is different from the analysis apparatus 1 in that the control unit 241 controls the processing operations of the magnetic flux collecting rod transfer unit 228 and the magnetic flux collecting rod cleaning unit 229 in place of the control unit 41, and the analysis unit 201 An analysis unit 243 that performs the same processing procedure as that of the unit 43 and analyzes the sample is provided. The reaction table 210 also transfers the reaction vessel 11 to the magnetic collection position by the magnetic rod transfer unit 228. Compared with the photometric unit 30, the photometric unit 230 has a configuration in which the permanent magnet 30c shown in FIG.

  As shown in FIG. 12, the magnetic flux collecting rod transfer unit 228 includes an arm 228b that is attached to the tip end of a magnetic flux collecting rod 228a that generates a magnetic field and that can freely move up and down and rotate. The magnetic flux collecting rod transfer unit 228 accommodates the specimen and the reagent and is located inside the reaction vessel 11 located at a predetermined magnetic flux collecting position of the reaction table 10 or at a washing position by the magnetic flux collecting rod washing unit 229 described later. Transport. Specifically, the magnetic flux collecting rod transfer unit 228 is a reaction vessel 11 that is transferred to the reaction table 10 according to a predetermined locus L2, and is located inside the reaction vessel 11 that is located at a predetermined magnetic collection position on the locus L1. Alternatively, the magnetic flux collecting rod 228a is transferred into the cleaning tank 229a of the magnetic flux collecting rod cleaning unit 229.

  As shown in FIG. 12, the magnetic flux collecting rod transfer unit 228 has a support 228c that is connected to the other end of the arm 228b and whose vertical line passing through the other end of the arm 228b is the central axis. The magnetic flux collecting rod transfer unit 228 includes a rotating mechanism 228d that rotates the support column 228c and an elevating mechanism 228e that moves the support column 228c up and down.

  The rotation mechanism 228d is connected to the pulley 2282d and fixed to the stationary part of the apparatus main body, the rotation motor 2281d, the pulley 2284d provided integrally with the column 228c, and the pulley 2282d and the pulley 2284d. A rotation belt 2283d. When the rotation motor 2281d rotates in a direction corresponding to clockwise or counterclockwise rotation, the rotation of the rotation motor 2281d is sequentially transmitted to the pulley 2282d, the rotation belt 2283d, and the pulley 2284d, so that the column 228c rotates about the central axis. Rotate clockwise or counterclockwise. By the rotation of the column 228c, the arm 228b to which the column 228c is connected rotates along the locus L2 as indicated by an arrow Y5.

  The elevating mechanism 228e is connected to the pulley 2282e and fixed to an immovable part inside the analyzer 1, an elevating motor 2281e, an elevating plate 2286e provided integrally with the lower end of the column 228c, and parallel to the column 228c. Thus, a screw shaft 2285e fixed to a stationary portion inside the analyzer 1 and provided with a pulley 2284e at the lower end, and a lifting belt 2283e suspended over the pulley 2282e and the pulley 2284e are provided. The screw shaft 2285e passes through the elevating plate 2286e, and the elevating plate 2286e and the screw shaft 2285e constitute, for example, a ball screw mechanism. When the lifting motor 2281e rotates in a direction corresponding to the ascending direction or the descending direction, the rotation of the lifting motor 2281e is sequentially transmitted to the pulley 2282e, the lifting belt 2283e, and the pulley 2284e, whereby the screw shaft 2285e rotates. As the screw shaft 2285e rotates, the elevating plate 2286e rises or falls, so that the column 228c rises or falls. As the column 228c is moved up and down, the arm 228b connected to the column 228c is also moved up and down as indicated by an arrow Y6.

  The magnetic flux collecting rod transfer unit 228 inserts the magnetic flux collecting rod 228a into the reaction container 11 in which the specimen, the magnetic reagent, and the labeling reagent are dispensed and the complex is generated. The magnetic flux collecting rod 228a inserted into the reaction vessel 11 generates a magnetic field in the reaction vessel 11, thereby attracting and collecting the magnetic particles in the reaction vessel 11 and the magnetic particles in the complex on the surface of the magnetic flux collecting rod 228a. Magnetize. Thereafter, the magnetic flux collecting rod transfer unit 228 raises the arm 228b, further rotates the arm 228b, and transfers the magnetic flux collecting rod 228a to the reaction vessel 11, thereby reacting the magnetic particles and the complex in the reaction vessel 11. Remove outside the container 11. Next, the magnetic flux collecting rod transfer unit 228 moves the magnetic flux collecting rod 228a onto the cleaning tank 229a in the magnetic flux collecting rod cleaning unit 229 described later, and then lowers the arm 228b to move the magnetic flux collecting rod 228a into the cleaning tank 229a. insert. As a result, the magnetic flux collecting rod 228a is cleaned in the cleaning tank 229a.

  As shown in FIG. 12, the magnetic flux collecting bar cleaning unit 229 is connected to the cleaning tank 229a, the side surface of the cleaning tank 229a, connected to the cleaning liquid tank and supplied with the cleaning liquid, and connected to the bottom surface of the cleaning tank 229a. And a drainage pipe 229c that is connected to a drainage tank and discharges cleaning liquid and the like, and cleans the magnetic flux collecting rod 228a.

  In the analyzer 201 according to the second embodiment, the magnetic collecting rod 228a is directly inserted into the liquid in the reaction vessel 11 to collect the magnetic particles and the composite in the reaction vessel 11 by the operation process of the magnetic collecting rod transfer unit 228. The magnetic particles and the composite are removed from the reaction vessel 11 by transferring the magnetism collection rod 228a to the outside of the reaction vessel 11 in a state of being collected on the surface of the magnetic rod 228a. As a result, the analyzer 201 removes the magnetic particles and the complex from the laser light irradiation region. The magnetic particles and composites collected on the surface of the magnetic flux collecting rod 228a are removed from the surface of the magnetic flux collecting rod 228a by the cleaning treatment of the magnetic flux collecting rod 228a by the magnetic flux collecting rod cleaning unit 229.

  In the analyzer 201 according to the second embodiment, the pre-reaction photometric process and the post-reaction photometric process are performed not on the same reaction vessel but on different reaction vessels 11 respectively. The analyzer 201 performs the pre-reaction photometric process on the pre-reaction photometric process reaction vessel 11 in advance for each specimen. The analyzer 201 uses the photometric value obtained by the pre-reaction photometric process as reference data for the same specimen. That is, when the analysis apparatus 201 is instructed to perform analysis processing a plurality of times for the same sample, the analyzer 201 performs the pre-reaction measurement process only once instead of performing the pre-reaction measurement process for each analysis process. Then, using the pre-reaction photometric value obtained by the pre-reaction measurement process as reference data, the difference value with the photometric value of each post-reaction photometric process of each analysis process is calculated to detect the concentration of the analyte in the sample.

  Specifically, the analysis process of the analyzer 201 will be described in detail. FIG. 13 is a flowchart illustrating a processing procedure of analysis processing performed by the analysis apparatus 201 illustrated in FIG. 11. As shown in FIG. 13, when an analysis process for a predetermined item is instructed for a predetermined sample, the analyzer 201 first applies a magnetic reagent, a sample, and a sample to the reaction vessel 11 for pre-reaction photometric processing. The labeling reagent is dispensed, and the magnetic particles before binding to the antigen are removed from the reaction container 11 before the binding reaction of the magnetic reagent, the antigen in the specimen and the gold particles proceeds, and then the gold particles before binding to the antigen. A pre-reaction measurement process (step S202) for measuring the intensity of the Raman scattered light due to is performed.

  Next, the analyzer 201 dispenses the magnetic reagent, the specimen, and the labeling reagent into the reaction container 11 different from the reaction container 11 used in the pre-reaction measurement process, and the magnetic particles in the magnetic reagent, After the complex is formed by the binding reaction of the antigen and the gold particle, the magnetic particle and the complex are removed from the reaction vessel 11 and then the intensity of the Raman scattered light by the gold particle that has not bound to the antigen as the analysis target. A post-reaction measurement process (step S204) for photometric measurement is performed.

  Next, in the analysis apparatus 201, the control unit 241 determines whether or not the measurement process for all the times specified for the sample to be analyzed has been completed (step S206). When the control unit 241 determines that the instructed measurement process has not been completed for the sample to be analyzed (step S206: No), the control unit 241 returns to step S204 and applies to the new reaction container 11. Perform measurement after the reaction. On the other hand, when the control unit 241 determines that the instructed measurement process has been completed for the sample to be analyzed (step S206: Yes), the analysis unit 243 performs the pre-reaction photometric process (step S202). ) And a photometric value in each post-reaction photometric process (step S204), a calculation process is performed to determine the concentration of the antigen 50 to be analyzed in the sample (step S208). Thereafter, the output unit 45 performs an output process for outputting the antigen concentration obtained by the analysis unit 43 (step S210), and ends the analysis process in the analyzer 1.

  Next, the pre-reaction measurement process shown in FIG. 13 will be described with reference to FIGS. 14 and 15. FIG. 14 is a flowchart showing a processing procedure of the pre-reaction measurement process shown in FIG. 13, and FIG. 15 is a diagram for explaining the pre-reaction measurement process shown in FIG.

  In the analyzer 201, first, as shown by arrows Y21 to Y23 in FIGS. 14 and 15 (1), the magnetic reagent dispensing unit 22 includes magnetic particles 51 in the reaction container 11a for pre-reaction measurement processing. And a sample dispensing process in which the sample dispensing unit 20 dispenses a sample containing the antigen 50 and the contaminant 55 in the pre-reaction measurement process reaction container 11a (step S220). Step S222) and a labeling reagent dispensing process (Step S224) in which the labeling reagent dispensing unit 25 dispenses the labeling reagent containing the gold particles 52 into the reaction container 11a for the pre-reaction measurement process. Then, the stirring unit 27 performs a stirring process for stirring the liquid in the reaction vessel 11a into which the magnetic reagent, the specimen, and the labeling reagent have been dispensed (step S226). Thereafter, the reaction table 210 transfers the reaction vessel 11a to a predetermined magnetic collection position before the binding reaction of the magnetic particles 51, the antigen 50, and the gold particles 52 proceeds.

  Then, the magnetic flux collecting rod transfer unit 228 performs a magnetic body removing process for removing the magnetic particles 51 from the reaction vessel 11a (step S228). Specifically, first, the magnetic flux collecting rod transfer unit 228 is shown in the reaction container 11a in which the magnetic particles 51, the antigen 50, the impurities 55, and the gold particles 52 are dispersed, as indicated by an arrow Y25 in FIG. The magnetic flux collecting rod 228a is inserted into The magnetic particles 51 are attracted and attracted to the surface of the magnetic collecting rod 228a by the magnetic field generated in the reaction vessel 11a by the magnetic collecting rod 228a. In this state where the magnetic particles 51 are collected on the surface of the magnetic flux collecting rod 228a, the magnetic flux collecting rod transfer unit 228 pulls up the magnetic flux collecting rod 228a as indicated by an arrow Y26 in FIG. The rod 228a is transferred out of the reaction vessel 11a. As a result, the pre-reaction measurement processing reaction container 11 a contains only the antigen 50, the impurities 55, and the gold particles 52 before binding to the antigen 50. That is, the magnetic particles before binding to the antigen 50 are removed from the laser light irradiation region. In the analyzer 201, since the magnetic particles 51 are collected by inserting the magnetic flux collecting rod 228a directly into the liquid in the reaction vessel 11a, efficient magnetic flux collecting processing is possible, and magnetic flux collecting by the magnetic flux collecting rod 228a is possible. A short time of about 10 seconds is sufficient.

  Next, the reaction table 10 transfers the reaction container 11 a containing only the antigen 50, the impurities 55, and the gold particles 52 before binding to the antigen 50 to the photometric position of the photometric unit 230. As shown in FIG. 15 (4), in the photometry unit 230, the laser light source 30 a irradiates the reaction vessel 11 a with the laser light Li, and the Raman spectrometer 30 b has the gold particles 52 before binding to the antigen 50. A pre-reaction photometry process (step S230) for measuring the intensity of the Raman scattered light Lfw is performed. In this pre-reaction photometric process, since the magnetic particles 51 are removed from the reaction vessel 11a, only the gold particles 52, the antigen 50 in the specimen, and the contaminants 55 are present in the irradiation region in the reaction vessel 11. Thus, the Raman scattered light corresponding to the total amount of the dispensed gold particles 52 is accurately measured. In FIG. 15 (4), the light condensing mechanism in the photometry unit 230 is not shown.

  Next, the photometric unit 230 outputs the photometric value in the pre-reaction photometric process to the control unit 241, whereby the pre-reaction measurement process ends. This pre-reaction measurement process need only be performed once for the same specimen. Note that the reaction vessel 11a used for the pre-reaction measurement process may be discarded or used for the analysis process again after washing.

  Next, the post-reaction measurement process shown in FIG. 13 will be described with reference to FIGS. 16 and 17. FIG. 16 is a flowchart showing a processing procedure of the post-reaction measurement process shown in FIG. 13, and FIG. 17 is a diagram for explaining the post-reaction measurement process shown in FIG.

  In the post-reaction measurement process, in the analyzer 201, as shown by arrows Y31 to Y32 in FIGS. 16 and 17 (1), a reaction container for post-reaction measurement process different from the pre-reaction measurement process reaction container 11a. In 11b, the same kind and the same amount of liquid as each liquid dispensed in each dispensing process in the pre-reaction measurement process are respectively dispensed. Specifically, the magnetic reagent dispensing unit 22 dispenses the same type and amount of the magnetic reagent dispensed in the pre-reaction measurement process into the reaction container 11b for the post-reaction measurement process. The sample dispensing process in which the sample (step S240) and the sample dispensing unit 20 dispense the same type and amount of sample as the magnetic reagent dispensed in the pre-reaction measurement process into the reaction container 11b for the post-reaction measurement process. (Step S242) and the labeling reagent dispenser 25 in which the labeling reagent dispensing unit 25 dispenses the same type and amount of labeling reagent as the magnetic reagent dispensed in the measurement process before the reaction into the reaction container 11b for the measurement process after the reaction. A note process (step S244) is performed. Therefore, as shown in FIG. 17 (2), in the reaction container 11b for the post-reaction measurement process, the same state as in the reaction container 11a after each liquid dispensing process in the pre-reaction measurement process shown in FIG. 15 (2). Thus, the magnetic particles 51, the antigen 50, the impurities 55, and the gold particles 52 may be regarded as being dispersed.

  And the stirring part 27 performs the stirring process which stirs the liquid in this reaction container 11b (step S246), and accelerates | stimulates reaction. Then, the analyzer 201 performs a reaction promoting process for promoting an antigen-antibody reaction that specifically binds the magnetic particles 51, the antigens 50, and the gold particles 52 in the reaction vessel 11b for a predetermined time (step S247), and FIG. As shown in (3), a complex 53 in which the magnetic particles 51, the antigens 50, and the gold particles 52 are specifically bound is generated in the reaction vessel 11b.

  Then, the magnetic flux collecting rod transfer unit 228 performs a magnetic body removal process for removing the magnetic particles 51 and the magnetic particles of the composite 53 from the reaction vessel 11b (step S248). Specifically, the magnetic flux collecting rod transfer unit 228 is the reaction vessel 11b in which the magnetic particles 51, the antigens 50, the impurities 55, the gold particles 52, and the complex 53 are dispersed, as in the magnetic substance removal process in the pre-reaction measurement process. Inside, as shown by an arrow Y35 in FIG. 17 (3), a magnetic flux collecting rod 228a is inserted. The magnetic particles 51 and the magnetic particles of the composite 53 are attracted and attracted to the surface of the magnetic collecting rod 228a by the magnetic field generated in the reaction vessel 11b by the magnetic collecting rod 228a. In this state where the magnetic particles 51 and the magnetic particles of the composite 53 are collected on the surface of the magnetic flux collecting rod 228a, the magnetic flux collecting rod transfer unit 228 moves the magnetic flux collecting rod as indicated by an arrow Y36 in FIG. 228a is pulled up, and this magnetism collecting rod 228a is transferred out of the reaction vessel 11b. As a result, the reaction container 11b for the post-reaction measurement process contains only the contaminants 55 and the gold particles 52 that have not bound to the antigen 50 that is the analysis target. In this magnetic body removal process, as in the magnetic body removal process in the pre-reaction measurement process, the magnetic particles 51 are collected by inserting the magnetic flux collecting rod 228a directly into the liquid in the reaction vessel 11b. As a result, it is possible to collect the magnetic flux by the magnetic flux collecting rod 228a in a short time of about 10 seconds.

  Next, the reaction table 10 transfers the reaction vessel 11b containing only the foreign particles 55 that have not been combined with the contaminant 55 and the antigen 50 that is the analysis target to the photometric position of the photometric unit 230. Then, as shown in FIG. 17 (5), in the photometry unit 230, the laser light source 30 a irradiates the reaction container 11 b with the laser light Li, and the Raman spectrometer 30 b does not bind to the antigen 50. A post-reaction photometric process (step S250) for measuring the intensity of the Raman scattered light Lfu is performed. In this post-reaction photometric process, the magnetic particles 51 of the complex 53 and the magnetic particles 51 that have not been bound to the antigen 50 are removed from the reaction vessel 11b. In the state where only the gold particles 52 not bound to the antigen 50 are present, the Raman scattered light corresponding to the gold particles 52 not bound to the antigen 50 can be measured accurately. In FIG. 17 (5), the light condensing mechanism in the photometry unit 230 is not shown.

  Next, the photometric unit 230 outputs the photometric value in the post-reaction photometric process to the control unit 241, whereby the post-reaction measurement process ends. As shown in FIG. 13, this post-reaction measurement process is executed for each instructed process content when an analysis process is instructed a plurality of times for the same sample. Note that the reaction vessel 11b used for the pre-reaction measurement process may be discarded or used for the analysis process again after washing.

  Next, the arithmetic processing shown in FIG. 13 will be described with reference to FIG. FIG. 18 is a flowchart illustrating a processing procedure of the arithmetic processing illustrated in FIG. As shown in FIG. 18, the analysis unit 243 acquires a pre-reaction photometric value in the pre-reaction measurement process of the sample to be analyzed (step S260). As the pre-reaction photometric value, a value obtained for a reaction vessel 11a different from the reaction vessel 11b in the post-reaction measurement process for the same specimen is used as a reference. Next, the analysis unit 243 acquires a post-reaction photometric value in the post-reaction measurement process of the sample (step S262). Then, the analysis unit 243 calculates a difference value between the acquired pre-reaction photometric value and the post-reaction photometric value (step S264). In the analyzer 201, as with the analyzer 1, the analysis unit 243 calculates the difference value between the pre-reaction photometric value and the post-reaction photometric value, thereby binding the antigen 50 and forming the complex 53. The intensity of the Raman scattered light by is indirectly acquired. Then, the analysis unit 243 calculates an analysis value corresponding to the difference value based on the difference value corresponding to the intensity of the Raman scattered light by the gold particles 52 that bind to the antigen 50 and constitute the complex 53. (Step S266), the calculation result is output to the control unit 241. Next, the analysis unit 243 determines whether or not the instructed calculation process has been completed for all the specified times (step S268). If the analysis unit 243 determines that the instructed calculation process has not been completed (step S268: No), the analysis unit 243 returns to step S260 and again sets the pre-reaction photometric value corresponding to the sample to be processed. Acquire. On the other hand, if the analysis unit 243 determines that the instructed processing for all the floors has been completed (step S268: Yes), the analysis processing ends.

  As described above, in the analyzer 201, similarly to the first embodiment, the magnetic particles 51 and the magnetic particles of the complex 53 are removed from the reaction vessel 11, and the arrival of the laser beam to the gold particles 52 is inhibited. In the absence of the above, the Raman scattered light corresponding to the total amount of the dispensed gold particles 52 and the Raman scattered light of the gold particles 52 not bonded to the analysis object can be accurately measured. Therefore, the analyzer 201 determines the difference between the intensity of the Raman scattered light corresponding to the total amount of the gold particles 52 before the reaction and the intensity of the Raman scattered light caused by the labeling substance that has not reacted with the analyte after the reaction. Therefore, it is possible to obtain an accurate value for the intensity of the Raman scattered light of the complex 53 itself, and the analysis accuracy can be remarkably improved.

  Further, in the analyzer 201, the magnetic particles 51 and the magnetic particles of the complex 53 are efficiently used because the magnetic particles 51 are collected by inserting the magnetic flux collecting rod 228a directly into the liquid of the reaction vessel 11. It can collect magnets well. Therefore, in the analysis apparatus 201, as in the first embodiment, the processing time of the entire analysis process can be shortened more reliably than before, and the number of sample processes per unit time can be increased to improve the processing capacity of the analysis process. Can be made.

  Furthermore, in the analyzer 201, when the analysis process is instructed a plurality of times for the same sample, the pre-reaction photometric process only needs to be performed once as a reference. For this reason, in the analyzer 201, it is not necessary to perform both the pre-reaction photometry process and the post-reaction photometry process for each analysis process. In other words, when the analysis apparatus 201 is instructed to perform analysis processing a plurality of times for the same sample, if the pre-reaction photometric process is performed once and the post-reaction photometric value is obtained, the subsequent processing for this sample is performed. It is only necessary to take a photometric value after the reaction. Therefore, the analyzer 201 does not necessarily need to perform the photometric process twice in order to obtain one analysis result, so that the total processing time itself for this specimen can be shortened.

  In the second embodiment, as in the first embodiment, the analysis unit 243 calculates the difference value between the pre-reaction photometric value and the post-reaction photometric value, thereby detecting the Raman scattered light of the complex labeling substance. The intensity was acquired, but by calculating the ratio between the pre-reaction photometric value and the post-reaction photometric value, the relative intensity of the Raman scattered light of the complex labeling substance was obtained, and the analyte concentration in the sample was detected. May be.

  In this case, as shown in the flowchart of FIG. 19, the analysis unit 243 acquires the pre-reaction photometric value (step S260) and acquires the post-reaction photometric value, similarly to step S260 and step S262 shown in FIG. Step S362). Then, the analysis unit 243 calculates a ratio between the pre-reaction photometric value and the post-reaction photometric value (step S364), calculates an analysis value for the calculated ratio (step S366), and sends the calculation result to the control unit 41. Output. Next, the analysis unit 243 determines whether or not the instructed arithmetic processing has been completed (step S368). When the analysis unit 243 determines that the instructed calculation process has not been completed (step S368: No), the analysis unit 243 returns to step S360, and again sets the pre-reaction photometric value corresponding to the sample to be processed. Acquire. On the other hand, if the analysis unit 243 determines that the instructed processing for all floors has been completed (step S368: Yes), the analysis processing ends.

  In the analyzers 1 and 201, the pre-reaction measurement process in which the concentration of the labeling reagent in the reaction container 11 is changed is performed in advance, and the pre-reaction photometric values for the concentrations of the respective labeling reagents are obtained. It is also possible to omit the pre-reaction measurement process by creating a calibration curve showing the relationship between the concentration and the pre-reaction photometric value. In this case, the analysis unit 243 may perform a calculation process by obtaining a photometric value corresponding to the concentration of the labeled reagent in the reaction container 11 from the calibration curve as the pre-reaction photometric value.

  In the analyzers 1, 201, the case where the labeling reagent is dispensed after the magnetic reagent has been described as an example. However, since there is no difference in the complex formation reaction depending on the reagent dispensing order, Reagents may be dispensed. Moreover, the apparatus layout of the analyzers 1 and 201 is an example, and the dispensing order is not restricted by this layout.

  In the analyzers 1 and 201, it is also possible to acquire a plurality of analysis results by performing a series of analysis processes using labeling reagents containing a plurality of types of labeling substances having different Raman peaks. .

DESCRIPTION OF SYMBOLS 1,201 Analyzer 2,202 Measuring mechanism 4,204 Control mechanism 10 Reaction table 11, 11a, 11b Reaction container 19 Specimen transfer part 19a Specimen container 19b Specimen rack 20 Specimen dispensing part 21 Magnetic reagent container 22 Magnetic reagent dispensing part 23, 26 Probe cleaning unit 24 Label reagent storage unit 25 Label reagent dispensing unit 27 Stirrer unit 30 Photometric unit 41, 241 Control unit 42 Input unit 43,243 Analysis unit 44 Storage unit 45 Output unit 50 Antigen 51 Magnetic particle 52 Gold particle 53 Complex 55 Contaminant 228 Magnetic flux collecting rod transfer unit 228a Magnetic flux collecting rod 229 Magnetic flux collecting rod cleaning unit

Claims (10)

A magnetic reagent containing magnetic particles that specifically bind to an analyte in a specimen, and a labeling substance that emits Raman scattered light whose surface is enhanced by irradiation with laser light and specifically binds to the analyte. In an analysis method for analyzing the sample using a labeling reagent containing
A dispensing step of dispensing the sample, the magnetic reagent and the labeling reagent into a reaction vessel;
Before the magnetic particles dispensed in the dispensing step and the labeling reagent are combined with the analyte in the specimen, the magnetic particles are removed from the laser light irradiation region in the laser in the reaction container. A pre-reaction photometry step of irradiating light and measuring Raman scattered light by the labeling substance in the reaction vessel;
A reaction promoting step of promoting a reaction in which a complex in which the magnetic particles, the analyte and the labeling substance are specifically bound in the reaction container is formed;
After the reaction promoting step, the laser beam is irradiated into the reaction container in a state where the magnetic particles that have not been combined with the analyte and the complex are removed from the laser light irradiation region, A post-reaction photometric step of measuring Raman scattered light by the labeling substance of
By calculating the difference or ratio between the photometric value in the pre-reaction photometric step and the photometric value in the post-reaction photometric step, the intensity of the Raman scattered light by the complex is obtained, and the Raman scattered light by the obtained complex is obtained. A calculation step for obtaining the concentration of the analyte in the specimen based on the intensity of
The analysis method characterized by including. In the pre-reaction photometry step, the magnetic particles are removed from the laser light irradiation region by generating a magnetic field in a reaction container into which the specimen, the magnetic reagent, and the labeling reagent are dispensed,
In the post-reaction photometric step, a magnetic field is generated in the reaction container in which the complex is generated, thereby removing the magnetic particles that have not been combined with the analyte and the complex from the irradiation region of the laser beam. The analysis method according to claim 1, wherein: In the pre-reaction photometric step, a magnet is brought close to an area other than the laser light irradiation area from the outside of the reaction container into which the specimen, the magnetic reagent, and the labeling reagent have been dispensed, whereby the magnetic particles are moved to the laser. Magnetize outside the light irradiation area,
In the post-reaction photometry step, a magnetic particle that has not been combined with the analyte and the composite are brought into proximity to a region other than the laser light irradiation region from the outside of the reaction vessel in which the composite is generated. The analysis method according to claim 1, wherein a body is collected outside the irradiation region of the laser beam.   The analysis method according to claim 3, wherein the magnet has a size capable of approaching at least a part of a bottom surface or a side surface of the reaction vessel.   The analysis method according to claim 3 or 4, wherein there are a plurality of the magnets. After the reaction in which the sample, the magnetic reagent, and the labeling reagent are dispensed in a reaction container for post-reaction measurement different from the reaction container for pre-reaction measurement in which the Raman scattered light is measured in the pre-reaction photometry step Further comprising a measuring dispensing step,
In the pre-reaction photometric step, a magnetic collecting rod that generates a magnetic field is inserted into the pre-reaction measurement reaction container into which the specimen, the magnetic reagent, and the labeling reagent have been dispensed, and the magnetic particles are collected into the magnetic collector. By removing the magnetic particles from the laser light irradiation region by transferring the magnetic rods to the outside of the reaction vessel for pre-reaction measurement in a state where the rods are magnetized,
In the post-reaction measurement dispensing step, the reaction promotion step promotes a reaction that generates the complex in a post-reaction measurement reaction container in which the specimen, the magnetic reagent, and the labeling reagent are dispensed,
In the post-reaction photometric step, the magnetic collecting rod is inserted into the reaction container for post-reaction measurement in which the complex is generated, and the magnetic particles that have not bound to the analysis object and the complex are The magnetic particles that have not been combined with the analysis object and the composite are transferred to the laser by transferring the magnetic collecting rod out of the reaction vessel for measurement after the reaction while collecting the magnetic flux on the surface of the magnetic collecting rod. The analysis method according to claim 1, wherein the analysis method is removed from the light irradiation region.   The analysis method according to claim 6, wherein the dispensing step and the pre-reaction photometric step are performed for each specimen. A magnetic reagent containing magnetic particles that specifically bind to an analyte in a specimen, and a labeling substance that emits Raman scattered light whose surface is enhanced by irradiation with laser light and specifically binds to the analyte. In an analyzer for analyzing the sample using a labeling reagent containing
Photometric means for irradiating the reaction vessel with laser light and measuring Raman scattered light from the labeling substance;
Removing means for removing the magnetic material from the laser light irradiation region in the reaction vessel;
A computing means for obtaining a concentration of the analyte in the specimen based on a photometric value obtained by the photometric means;
With
The removal means includes a pre-reaction removal process for removing the magnetic particles from the laser light irradiation region before the magnetic particles and the labeling reagent are combined with the analyte in the specimen, and the composite by the reaction means. A post-reaction removal process that removes the magnetic particles that did not bind to the analyte after the body was generated and the complex from the irradiation region of the laser beam,
In the photometric process, the marker in the reaction container is irradiated with the laser light in a state where the magnetic particles are removed from the laser light irradiation region by the pre-reaction removal process by the removing means. The pre-reaction photometric process for measuring Raman scattered light by the substance and the post-reaction removal process by the removing means remove the magnetic particles and the complex that are not bound to the analyte from the irradiation region of the laser beam. In this state, the reaction vessel is irradiated with the laser light to perform post-reaction photometric processing for measuring Raman scattered light from the labeling substance in the reaction vessel,
The computing means obtains the intensity of Raman scattered light by the complex by computing the difference or ratio between the photometric value in the pre-reaction photometric process and the post-reaction photometric process by the photometric means, An analyzer that determines the concentration of the analyte in the specimen based on the acquired intensity of Raman scattered light from the complex.   The removing means removes the magnetic particles from the laser light irradiation region by generating a magnetic field in a reaction container in which the specimen, the magnetic reagent, and the labeling reagent are dispensed in the pre-reaction removal process, In the post-reaction removal process, by generating a magnetic field in the reaction container in which the complex is generated, the magnetic particles that have not been combined with the analyte and the complex are removed from the laser light irradiation region. The analyzer according to claim 8.   An analysis program causing an analysis apparatus to execute the analysis method according to claim 1.

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