JP5160461B2 - Automatic analyzer - Google Patents

Description

  The present invention relates to an automatic analyzer for measuring components in a biological sample such as blood, and more particularly to an automatic analyzer capable of simultaneous measurement of a plurality of items.

  As a method for measuring the concentration of an antigen consisting of a protein in blood, a method utilizing an antigen-antibody reaction is known. An antibody that binds to the antigen in the sample and a label are added, and the signal from the label of the generated immune complex is counted, whereby the antigen is quantified and qualitatively measured.

  As a method for counting the labeling body, a method is mainly used in which a luminescent labeling body is used and the amount of luminescence is converted to be equal to the antigen concentration.

  As an example, Patent Document 1 discloses a method of measuring and emitting a label using a chemical reaction, and Patent Document 2 discloses a method of measuring by electrochemiluminescence.

  Patent Document 3 discloses a method of using a fluorescent substance as a label, irradiating irradiation light to excite the fluorescent substance, and measuring the light emission amount of the fluorescent substance using this excitation as a trigger.

  In the above-described method, a series of measurement operations are automated, and it is widely used as an automatic analyzer for pharmaceutical development research and clinical examination.

  At present, the need for this method in clinical tests is further expanded, and not only blood viruses and hormones, but also important test items for infectious diseases such as HAV, HBV, and HIV, and diseases such as cancer and rheumatism are measured. It is becoming.

  In a conventional automatic analyzer, a sample that has undergone an antigen-antibody reaction is caused to flow on a flow path including a flow cell in a measurement unit, and a voltage is applied inside the flow cell to emit light from a labeled body. ) Etc. Therefore, only one antibody concentration can be measured in one analysis, and many items of measurement require a lot of time and labor.

  In view of the above problems, a system capable of simultaneously measuring a plurality of types of antigen-antibody reactions by applying DNA chip technology is known. The number of items to be measured is applied on the analysis chip in advance for the antibody to bind to the analyte and the label. For example, a chip coated with a plurality of items in a line corresponding to the measurement items is prepared.

  By dispensing the biological sample to be measured on this chip and the corresponding dedicated reagent, the substance to be measured binds to each antibody in the analysis chip and the label in the reagent. Can be analyzed.

  This system has made it possible to perform analysis with a reduction in the time required for measurement and the required amount of sample compared to the conventional method.

It has become possible to measure a low concentration region (about 10 ⁻⁹ to 10 ⁻¹⁵ gcm ⁻³ ) as typified by a cancer marker.

JP 2003-50204 A Japanese National Patent Publication No. 11-507726 Japanese Patent Laid-Open No. 10-319011

  When simultaneously measuring images of luminescent signals from a plurality of items, antigen-antibody reactions of a plurality of measurement objects proceed in parallel in the same analysis chip. Since the analysis chip of this method is not partitioned for each analysis or analysis item, the obtained image data is a disturbance factor, that is, other analysis items to be measured at the same time, foreign substances that enter the analysis container during the measurement operation, It is very susceptible to defects from each mechanism.

  For example, when there is a difference in the light emission amounts of the analysis items measured next to each other, the light emission from the analysis item with the large light emission amount may interfere with the measurement of the analysis item with the small light emission amount.

  In addition, foreign matter mixed in the analysis chip is measured by reflecting / scattering fluorescence excitation light, and the amount of luminescence not proportional to the amount of antigen in the biological sample is detected due to insufficient BF separation of the analysis chip. There are things.

  An object of the present invention is to provide a system for detecting a measurement defect by extracting a characteristic signal pattern from an image measurement result, and to perform a recovery operation and remeasurement by feeding back the result of detecting an abnormality to the apparatus. Thus, an object of the present invention is to provide an automatic analyzer that can accurately analyze a plurality of items from a single measurement result.

  The configuration of the present invention for achieving the above object is as follows.

Assumed in advance in an automatic analysis device equipped with an analysis chip that analyzes multiple items in parallel on a biological sample in one plane and a measuring means that simultaneously measures signals in a plurality of analysis items on the analysis chip Storage means for storing the factor and a measurement abnormality result caused by the factor in association with each other, and a measurement specific to the associated abnormality factor from the measurement abnormality result stored in the storage unit Extraction means for extracting an abnormal pattern, determination means for comparing the actual measurement result with the measurement abnormal pattern to determine the presence / absence of the measurement result , a biological sample, a test reagent, and a luminescent label are separated on the analysis chip. Dispensing means for dispensing, and the reaction of the biological sample dispensed in the analysis chip by the dispensing means, the test reagent, and the label are uniformly promoted A response enhancing means, and separation and removal means for removing the unwanted substances from the analysis chip, when detecting an abnormal measured by comparison of the measurement abnormality pattern from association with abnormal factors stored in said storage means An abnormality cause estimating means for estimating at least one of an abnormality content and an abnormality cause, and the abnormality cause is detected by at least one of the reaction promoting means, the separation removing means, and the measuring means by the abnormality cause estimating means. An abnormality recovery means for automatically performing an abnormality recovery operation on the device mechanism when it is estimated that the device mechanism is present, and the analysis chip wall surface is provided with at least one of a biological sample, a test reagent, and a label binding agent is applied, the measuring means serves to image measuring the light emission from the label in the analysis chip, the abnormality recovery means, before The cleaning liquid discharge direction of the separation / removal means, the cleaning liquid discharge rate, the cleaning liquid discharge speed, the distance to the measurement target analysis chip, the relative position of the measurement means relative to the measurement target analysis chip, the relative angle of the measurement means, and the measurement means An automatic analyzer characterized by adjusting at least one of the distances.

  Here, factors that cause measurement abnormalities are assumed to be, for example, foreign matter mixed in the analysis chip, malfunctions in the components of the automatic analyzer, and measurement errors when measuring items with large and small amounts of luminescence at the same time. Therefore, the tendency of measurement abnormality to these factors is extracted from a plurality of measurement results whose influence is remarkable.

  As a measurement method, it is assumed that fluorescence emission generated by irradiating the analysis chip with excitation light is measured from above the chip and analysis items are judged from the image data and abnormalities are diagnosed. Instead, other optical measurement methods such as chemiluminescence and electrochemiluminescence may be used.

  Further, in order to quantitatively determine the presence or absence of abnormality, a threshold value that becomes a boundary of abnormality analysis may be set, and this value may be stored in association with the abnormality factor. The threshold values include the average signal value, signal standard deviation, amplitude and frequency of fluctuation of signal amount when measuring all reaction areas of the analysis chip, as well as the color of emitted light and magnitude of emitted light. , The evaluation amount such as the shape of light emission is extracted as a threshold value, and the abnormality is determined by comprehensively comparing the plurality of threshold values with the evaluation amount obtained from the measurement result.

  When a measurement abnormality occurs, the cause of the abnormality is estimated by comparing the stored abnormality factor with the abnormality pattern. When the cause of the abnormality can be identified, it is desirable to have a function of automatically performing an operation for eliminating the cause of the abnormality or repairing the defect.

  In addition, when the cause is resolved, it is better to have a function of automatically performing remeasurement.

  In an automatic analyzer that can measure the amount of light from multiple analysis items at the same time, the abnormal state of the analysis item that caused measurement abnormality due to an external factor is detected appropriately, the cause of the abnormality is quickly investigated, and an appropriate Recovery operation can be executed promptly.

The block diagram explaining an Example. The figure explaining the substance adhering to the analysis container. The figure explaining attachment and measurement of a labeled body. The figure explaining a detection mechanism.

  Embodiments of the present invention will be described below with reference to the drawings.

  First, an embodiment having a function of detecting an abnormality of a mechanism provided in the system is shown in a block diagram of FIG. In this embodiment, as shown in FIG. 2, a substance 101 having a property of binding to a label is attached to the bottom surface of the analysis container 100. A fluorescent material 202 having a property of emitting light 206 and 307 using irradiation 205 and 304 as a trigger is used as the labeling body. The photometric means 203 and 309 include an image sensor such as a CCD, CMOS, and PMT (photomultiplier tube), and the irradiation light sources 204 and 301 include an LED having a center wavelength of the spectrum at the excitation wavelength of the marker, a halogen lamp, or the like. Use a white light source.

  The system configuration is shown in a block diagram in FIG. This system includes a reagent and sample dispensing mechanism 10, a stirring mechanism 11, a cleaning mechanism 12, a detection mechanism 13, an image data acquisition unit 14, an image data storage unit 15, an image data analysis calculation unit 16, a database 17, and a mechanism abnormality determination. It comprises an arithmetic unit 18, an alarm output function 19, and a mechanism control board 20.

  First, the analysis container 100 is fixed to the stage 102 of the dispensing mechanism 10, a test reagent is mixed in the container, and is combined with the substance 101 applied to the bottom surface of the analysis container. Next, the analysis container 100 is carried to the stirring mechanism 11. In this case, stirring is performed so that the substance and the label are uniformly and sufficiently combined, whereby the substance in the analysis container is sufficiently mixed. As a stirring method, there is a non-contact method such as air injection or ultrasonic vibration, or a contact method using a stirring rod.

  After stirring, the analysis container 100 undergoes a cleaning process in which the unreacted test reagent and the non-measurement object that have not been combined with the substance applied to the bottom surface of the container are removed from the analysis container by the cleaning mechanism 12. Thereafter, the light quantity is measured by the detection mechanism 13.

  The data acquired by measuring the amount of light is in the form of image data in which one signal amount corresponds to each lattice point on the plane coordinates. After the data is acquired by the image data acquisition unit 14, it is transmitted to the image data storage unit 15. After the transmission, the image data analysis calculation unit 16 analyzes the acquired data, and determines the signal intensity distribution of the light emitted from the label, the average signal amount of the entire analysis container, the standard deviation value, the fluctuation frequency, and the like. It is calculated according to the algorithm.

  Next, the mechanism abnormality determination calculation unit 18 compares the current measurement result with the abnormality measurement result recorded in the database 17. Here, the database 17 is an analysis performed under a situation where a known content abnormality or failure has occurred in each mechanism (stirring mechanism 11, cleaning mechanism 12, flow path system, etc.) provided in the system, and analysis of each result. Alternatively, verification by numerical simulation is performed, a unique tendency appearing on the image data is extracted, and stored in advance in association with the abnormal content of the mechanism.

  For example, if there is some problem in the stirring mechanism 11 and the stirring target area has deviated from the reaction area, the signal amount distribution of the image data acquired by the analysis becomes non-uniform. If the cleaning mechanism 12 has some problems and is not sufficiently cleaned, the entire signal amount becomes large due to light emission from the marker that cannot be completely washed and reflected light from other foreign matters, and the signal amount distribution. Such a feature that a local light emitting region exists in the region appears. In addition, if there is something wrong with the flow path system for flowing the cleaning liquid and the flow of liquid is hindered, there will be a difference in the discharge speed of the cleaning liquid, resulting in a spotted structure in the signal amount distribution, etc. A feature appears in the image data according to the problem.

  In order to quantitatively evaluate the above characteristics with respect to the measurement results, first, based on a predetermined algorithm, the average value, standard deviation value, and signal amount that is multiple times the standard deviation from the average value over the entire reaction region of the analytical container The evaluation quantity (physical quantity) such as the distribution or area of the area having the amplitude, the amplitude and frequency with respect to the fluctuation of the signal quantity is calculated. For these calculated evaluation quantities (physical quantities), a threshold value is set as a reference value for determining whether or not there is a malfunction. Since whether or not a defect has occurred is determined by comprehensively judging the plurality of evaluation amounts, the threshold value is set as one set for each of the plurality of types of evaluation amounts for each defect content. It is a value to be determined. These pieces of information are stored in advance in a database.

  The mechanism abnormality determination calculation unit 18 compares the evaluation amount calculated by the image data analysis calculation unit 16 with the threshold value recorded in the database 17 to check whether an unsteady signal pattern is included. At this time, if the presence of an unsteady signal pattern is confirmed, and it is determined that the cause is due to an abnormality in the mechanism, an alarm is output from the alarm output unit 19, and the contents of the malfunction that has occurred and the mechanism in which the malfunction has occurred To the user.

  It is also possible to feed back the determination result to the mechanism system via the mechanism control board 20 and to provide a function for repairing the defect by the apparatus itself. For example, if an abnormality occurs in the cleaning mechanism 12, the direction of discharge of the cleaning liquid used for cleaning, the amount of discharge, the speed of discharge, the relative position with respect to the discharge analysis container, the relative distance with respect to the discharge analysis container, etc. Adjust and restore to ensure sufficient agitation. If it is determined that the flow path system is abnormal, the flow path is cleaned.

  By the above method, it becomes possible to detect anomalies using the analysis result of the measurement data, and in the past, this method has the effect of greatly reducing the labor required for investigating defects. As a result, cost reduction is realized. In addition, by providing a function that allows the device itself to perform self-maintenance as necessary, the device can always be kept in a constant state, and as a result, stable analysis results can be provided. .

  Secondly, an embodiment having the function of judging the analysis condition at the time of measuring the light quantity from the analysis result, performing self-maintenance based on the judgment result, adjusting the analysis parameter as necessary, and performing the re-measurement is shown. . In this embodiment, a plurality of types of antigen-antibody reactions are performed in the same analysis container. A label for recognizing the reaction position is attached to the position where the antigen-antibody reaction is performed. A fluorescent material 202 is used as the label.

  First, a blood sample and a test reagent are mixed in an analysis container, and an antibody attached to the analysis container 100 is reacted with an antigen to be measured in the blood sample to form a complex. Next, stirring is performed and the reaction solution is sufficiently mixed. After the reaction is sufficiently performed, a test reagent mainly composed of a fluorescent label is mixed in the analysis container, and the fluorescent label is attached to the complex. After the fluorescent dye is sufficiently attached to the complex, the analysis container 100 is carried to the stages 201 and 305 through the process of removing the unreacted test reagent and the non-measurement object, and the amount of luminescence is measured.

  The light emitted from the analysis container includes the light 206 and 307 emitted from the label 202 attached to the complex, the reflected light from the foreign matter mixed during the analysis, and the unreacted fluorescent label that cannot be removed in the removal process. The amount of signal obtained by adding the noise signal amount peculiar to the sensor to these is acquired as a measured value. In order to accurately obtain an effective signal for concentration measurement caused by the antigen-antibody reaction, it is necessary to separate light emitted from the complex from the acquired signal amount.

  Therefore, consider the following means.

  First, as described in Example 1, an image obtained as reflected light of a foreign substance expected to be mixed and an image emitted from a fluorescent label to be used are analyzed in advance, and each spectrum, shape, and size (light emission) are analyzed. Range), signal amount, and the like are calculated and stored in the database 17 in association with the type of foreign matter.

  Image data acquisition unit 14 Image data acquired by light quantity measurement and stored in the image data storage unit 15 is calculated by the image data analysis calculation unit 16 using a predetermined algorithm, and the region in which the antigen-antibody reaction has been performed is calculated. Evaluation amounts such as coordinates, luminescence from the complex, luminescence from other than the complex, luminescence from the background region where no antigen-antibody reaction has occurred, and standard deviation values are calculated.

  Among the calculated values, for light emitted from other than the complex, the information stored in the database 17 is compared with the signal amount, shape, spectrum of light emission, and size to determine whether the light is emitted from the label.

  If it is determined that the light is from the label, it is likely that the label contained in the unreacted test reagent could not be washed completely, or the label was peeled off from the complex. Automatically check for abnormalities in function and cleaning function. Perform re-measurement as required by the user. At the time of re-measurement, the analysis conditions are optimized by adjusting the shape of the outlet for discharging air and cleaning liquid and controlling the direction, amount and speed of discharge.

  On the other hand, if it is determined that the light is not emitted from the sign, there is a high possibility that it is reflected light from the foreign matter mixed during the analysis measurement. Perform a remeasurement.

  Further, regarding the light emission amount from the background region, the average signal amount is calculated for four predetermined regions on the front, rear, left and right sides of the analysis container. The average signal amounts of these four areas are compared. If there is a difference in the average value, there may be uneven irradiation of irradiation light or there may be a tilt in the arrangement of the analysis container. These pieces of information are transmitted to the mechanism control board 20, and the rotation shaft 306 is rotated to adjust the angle at which the analysis container is disposed, and the entire analysis container is disposed so as to obtain uniform irradiation. Alternatively, the irradiation angle of the LED 204 is adjusted so that uniform irradiation can be obtained over the entire analysis container, and remeasurement is performed as necessary. At this time, depending on the case, stirring for uniform mixing of biological sample and label, adjustment of dispensing volume, control of stirring duration, etc., and re-measurement by selecting optimal parameters for analysis .

  In this way, the light emission from the foreign matter or the light emission from other than the complex is used to detect the malfunction of the mechanical system and the components of the apparatus. Moreover, it is possible to provide a more reliable analysis result by performing feedback and performing remeasurement as necessary.

DESCRIPTION OF SYMBOLS 10 Dispensing mechanism 11 Agitation mechanism 12 Washing mechanism 13 Detection mechanism 14 Image data acquisition part 15 Image data storage part 16 Image data analysis calculation part 17 Database 18 Mechanism abnormality determination calculation part 19 Alarm output part 20 Mechanism control board 100 Analysis container 101 Label An example of an assembly of substances that easily bind to a body 102 An example of a stage in a dispensing mechanism 201 An example of a stage in a detection mechanism 202 A substance to which a label is attached 203 An example of an image sensor 204 An irradiation light source 205 An irradiation light (excitation light)
206 Storage Box 302 with Irradiation Light Source Arranged in Emission Light 301 Lens 303 for Condensing Irradiation Light Tube 304 in Optical Path of Irradiation Light Irradiation Light (Excitation Light for Fluorescent Substance)
305 Operation stage 306 for fixing the analysis container in the detection unit Rotating shaft 307 Radiation light 308 from the label (fluorescent substance) 308 Optical path tube 309 for collecting the radiation light Box for storing the image sensor

Claims (5)

An analysis chip that analyzes multiple items in parallel in one plane for a biological sample;
Measuring means for simultaneously measuring signals in a plurality of analysis items on the analysis chip;
In an automatic analyzer equipped with
Storage means for storing the factor and the measurement abnormality result caused by the factor in association with each other, with respect to the factor causing the measurement abnormality assumed in advance,
Extraction means for extracting a measurement abnormality pattern specific to the associated abnormality factor from the measurement abnormality result stored in the storage means;
A determination means for comparing the actual measurement result with the measurement abnormality pattern to determine whether there is an abnormality in the measurement result;
A dispensing means for dispensing a biological sample, a test reagent and a luminescent label on the analysis chip;
Reaction accelerating means for uniformly accelerating the reaction of the biological sample, the test reagent and the label dispensed in the analysis chip by the dispensing means;
Separation / removal means for removing unnecessary substances from the analysis chip;
When a measurement abnormality is detected by comparison with the measurement abnormality pattern, an abnormality cause estimation unit that estimates at least one of an abnormality content and an abnormality cause from an association with an abnormality factor stored in the storage unit;
When it is estimated by the abnormality cause estimating means that the abnormality factor is present in at least one of the reaction promoting means, the separation removing means, and the measuring means, an abnormality is automatically made to the device mechanism. An abnormality recovery means for performing a recovery operation,
A substance that binds to at least one of a biological sample, a test reagent, and a label is applied to the wall of the analysis chip,
The measuring means measures the light emission from the label in the analysis chip,
The abnormality recovery means includes a cleaning liquid discharge direction, a cleaning liquid discharge amount, a cleaning liquid discharge speed, a distance to the measurement target analysis chip, a relative position of the measurement means with respect to the measurement target analysis chip, and the measurement means. An automatic analyzer that adjusts at least one of the relative angle of the distance and the distance of the measuring means. The automatic analyzer according to claim 1, wherein
An air stirring mechanism is provided as the reaction promoting means,
Equipped with a stirring air discharge mechanism that discharges air to the analysis chip and is variable in size and shape,
The abnormality recovery means adjusts at least one of the air discharge direction, the air discharge amount, the air discharge speed, the air discharge time, the relative position with respect to the measurement target analysis chip, and the distance with respect to the measurement target analysis chip of the stirring air discharge mechanism. The automatic analyzer characterized by doing. The automatic analyzer according to claim 2,
The abnormality recovery means controls the dispensing amount of at least one of a biological sample, a test reagent, and a label with respect to the dispensing means. The automatic analyzer according to claim 1, wherein
The abnormality recovery means is an automatic analyzer characterized in that a biological sample is agitated by the reaction promoting means before the retest function performs remeasurement. The automatic analyzer according to claim 1, wherein
An automatic analyzer having a function of selecting whether or not to perform re-measurement from the user side after performing an abnormality recovery operation by the abnormality recovery means.

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