JP2017015671A - System for quantifying organism related substance - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a system for quantifying an organism related substance, that allows an organism related substance to be quantified without practically controlling the processing temperature.SOLUTION: The system for quantifying an organism related substance includes: a processing area in which predetermined processing is carried out for quantifying an organism related substance in a solution; a processing accelerating tool for accelerating the processing in the processing area; a temperature measuring unit for measuring an actual processing temperature in the processing area at the time of processing; a control unit for setting processing conditions excluding the processing temperature on the basis of a comparison between the processing temperature and a predetermined reference temperature, and operating the processing accelerating tool on the basis of the set processing conditions; and a measuring unit for measuring the organism related substance in order to quantify the organism related substance, after operating the processing accelerating tool. The control unit, when the organism related substance is set at the same concentration, sets the processing conditions so that an actual measured value at the processing temperature is practically the same as a reference measured value of the organism related substance at the reference temperature.SELECTED DRAWING: Figure 1

Description

  The present invention relates to a biological material quantitative system, and more particularly to a biological material quantitative system capable of quantitatively determining biological material regardless of temperature.

  An immunoassay such as ELISA (Enzyme Linked Immunosorbent Assay) is used for the quantification of the biological substance. In the immunoassay, since the reaction rate differs depending on the treatment temperature during the treatment such as the antigen-antibody reaction, it was necessary to maintain the treatment temperature at a reference temperature such as 37 ° C. using a thermostat to obtain a stable measurement value. . Furthermore, in addition to maintaining the reference temperature in this way, the biologically relevant substance is quantified after the treatment is performed with all treatment conditions such as stirring and time fixed.

  In such an immunoassay, a well plate having a large number of wells is used. An antigen-antibody reaction is performed in a state where a solution containing a biological substance is contained in a well. By heating the periphery of these wells with a thermostat such as a heat block, the processing temperature has been maintained at a reference temperature such as 37 ° C.

  On the other hand, the quantitative system described in International Publication No. 2010/122990 has been proposed to perform a treatment in a tube by sucking a solution containing a biological substance from the well into the tube.

International Publication No. 2010/122990 Pamphlet

  Conventionally, in an immunoassay, when performing an antigen-antibody reaction in a well in order to quantify biological substances, it was necessary to fix all treatment conditions including temperature. In order to heat and maintain the processing temperature from room temperature to a reference temperature such as 37 ° C., it takes time and strict management is difficult. In addition, in immunoassay, it may be necessary to leave the treatment at room temperature for a few hours to a whole day and night, and it takes time until the determination. In addition, there is a concern that the manufacturing cost of the apparatus may increase by providing a thermostat such as a heat block.

  Furthermore, since the quantitative system described in International Publication No. 2010/122990 does not include a heating device that heats the tube to a reference temperature, the solution containing a biological substance is moved when the solution is moved between the tube and the well. There has been a risk that the processing temperature of this would change from the reference temperature.

  Therefore, an object of the present invention is to provide a novel system for quantifying a biological substance, which is capable of quantifying the biological substance. Another object of the present invention is to provide a biological material quantitative system capable of quantitatively determining biological material without substantially controlling the processing temperature.

  The present invention is as follows. (Aspect 1) A system for quantifying a biological substance, a processing region for performing a predetermined process for quantifying the biological substance in a solution, a processing accelerator for promoting the processing in the processing region, A temperature measurement unit that measures the processing temperature of the processing region during the processing, and sets processing conditions other than the processing temperature based on a comparison between the processing temperature and a predetermined reference temperature, and the set processing conditions A control unit that operates the processing accelerator based on the above, and a measurement unit that measures the biological substance to quantify the biological substance after the processing accelerator is operated, and the control unit includes: When the biological substance has the same concentration, the measurement value at the processing temperature substantially corresponds to or substantially the same as the reference measurement value of the biological substance at the reference temperature. Said To change the physical conditions, system.

  (Aspect 2) In the system for quantifying a biological substance according to Aspect 1, the processing conditions include a time during which the processing accelerator performs the processing, a speed at which the processing accelerator mixes the solution, and the processing accelerator The number of times of mixing the solution, or the amount of liquid in which the processing accelerator is mixed with the solution, or any combination of the plurality. (Aspect 3) In the system for quantifying a biological substance according to Aspect 1 or 2, the processing region moves between one or a plurality of tubes capable of sucking and discharging the solution and the solution between the tubes. A system with one or more wells to accommodate. (Aspect 4) The system for quantifying a biological substance according to Aspect 3, wherein the processing promoting tool includes a pump for sucking and discharging the solution with respect to the tube. (Aspect 5) The system for quantifying a biological substance according to any one of Aspects 1 to 4, wherein the reference temperature is room temperature or body temperature. (Aspect 6) In the system for quantifying a biological substance according to any one of the aspects 1 to 5, the measurement unit measures fluorescence of a substance combined with the biological substance.

  (Aspect 7) The system for quantifying a biological substance according to any one of Aspects 1 to 6, wherein the treatment includes an antigen-antibody reaction or nucleic acid hybridization. (Aspect 8) In the system for quantifying a biological substance according to any one of Aspects 1 to 7, the control unit has a predetermined temperature range in which the processing temperature measured by the temperature measurement unit includes the reference temperature. The processing conditions are set when included in the system. (Aspect 9) The system for quantifying a biological substance according to Aspect 8, wherein the predetermined temperature range is a range from about 20 ° C. to about 30 ° C., or a range from about 20 ° C. to about 37 ° C. . (Aspect 10) In the biological substance quantification system according to Aspect 8, the predetermined temperature range is 15 ° C. ± 5 ° C., 20 ° C. ± 5 ° C., 25 ° C. ± 5 ° C., or 30 ° C. ± 5 ° C. ,system.

  (Aspect 11) In the system for quantifying a biological substance according to any one of Aspects 1 to 10, a first substance in which a substance capable of binding to a biological substance to be measured is fixed in the reaction region. Microparticles, second microparticles in which the biological substance is fixed in a predetermined amount, and third microparticles for negative control in which the biological substance is treated so as not to be bound are arranged in a row ,system. (Aspect 12) The system for quantifying a biological substance according to Aspect 11, wherein the first microparticle is a microparticle for correcting measurement data of the biological substance. (Aspect 13) The system for quantifying a biological substance according to Aspect 12 or 13, wherein the second microparticle is a plurality of microparticles to which different amounts of the biological substance are immobilized. (Aspect 14) The system for quantifying a biological substance according to Aspect 13, wherein the plurality of microparticles are microparticles for preparing a calibration curve. (Aspect 15) The living body related substance determination system according to any one of Aspects 11 to 14, further comprising a light-shielding member between the first to third microparticles. ,system. (Aspect 16) In the system for quantifying a biological substance according to any one of aspects 11 to 14, the measurement unit detects a signal emitted from the microparticle in the tube, and the control unit is created in advance. A system for correcting the data of the detected signal with reference to the calibration curve and quantifying the biological substance based on the corrected data. (Aspect 17) In the system for quantifying a biological substance according to any one of aspects 11 to 14, the measurement unit detects a signal emitted from a microparticle in the tube, and the control unit detects the signal. A system for preparing a calibration curve based on the obtained signal and quantifying a biological substance with reference to the created calibration curve.

  According to the present invention, by changing the processing conditions based on the processing temperature, strict temperature management is not required, the time until the temperature is constant, the incubator necessary for the constant temperature is not necessary, and the system is simplified. It becomes possible.

It is a graph which shows the change of the detection intensity I with respect to the process temperature T used as the premise of the fixed_quantity | quantitative_assay system of this invention. It is the schematic of the tube used for the fixed_quantity | assay system of this invention. It is a block diagram of the fixed_quantity | quantitative_assay system of this invention. It is a perspective view of the measurement mechanism which measures the emitted light intensity by each bead of FIG. It is explanatory drawing which illustrates an antigen antibody reaction about fixed_quantity | assay operation of this invention. It is explanatory drawing of the measured value correction | amendment method using the emitted light intensity and density | concentration of a correction bead.

  A biological material quantitative system according to an embodiment of the present invention will be described with reference to the drawings. As described in the background art, a reaction such as an antigen-antibody reaction for separating a biological substance depends on a processing temperature. Therefore, even if the same processing is performed at the same time if the processing temperature is different, the detection intensity of fluorescence or the like related to the separated biological material is different. The processing temperature is the temperature of the processing area where processing is performed. The processing temperature is measured during and / or before and after processing in either the processing system including the processing region or the room in which the processing system is installed.

  FIG. 1 is a diagram showing a change in detection intensity I such as fluorescence indicating the concentration of a biological substance with respect to the processing temperature T when conditions other than the processing temperature are the same. According to the inventor's knowledge, it has been clarified that the detection intensity exhibited by the biological substance having the same concentration is linearly dependent or correlated with the processing temperature within a certain temperature range. More specifically, as shown by the black dots in FIG. 1, within a certain temperature range, the detection intensity indicated by the biological substance having the same concentration can be approximated so as to be proportional to the processing temperature. The constant temperature range is, for example, a range from room temperature to body temperature, and can be preferably a range from about 20 ° C. to about 30 ° C., or a range from about 20 ° C. to about 37 ° C. Alternatively, the constant temperature range may be preferably 15 ° C. ± 5 ° C., 20 ° C. ± 5 ° C., 25 ° C. ± 5 ° C., or 30 ° C. ± 5 ° C.

  In the quantification system of the embodiment of the present invention, when quantifying a biological substance, the control unit controls the number of operations, the operation time, and the operation speed of the processing accelerator based on the relationship between the temperature and the detected intensity in FIG. And / or change the program that defines the processing conditions such as the amount of solution to be mixed (suctioned / discharged), and execute the processing. There are the following two cases of specific processing condition changes. First, when the processing temperature is lower than the reference temperature, the control unit increases the actual number of operations with respect to the reference operation number of the processing accelerator, or actually increases with respect to the reference operation time of the processing accelerator. Or increase the actual operating speed with respect to the standard operating speed of the processing accelerator, or the actual liquid volume aspirated / discharged by the processing accelerator such as a dispenser. The amount of solution sucked and discharged can be increased, or any combination thereof can be implemented. Secondly, when the processing temperature is higher than the reference temperature, the control unit reduces the actual number of operations with respect to the reference operation number of the processing accelerator, or the actual time with respect to the reference operation time of the processing accelerator. The actual operation speed is reduced with respect to the reference operation speed of the processing accelerator, or the actual operation speed is reduced with respect to the reference liquid amount of the solution sucked and discharged by the processing accelerator such as a dispenser. The amount of solution sucked and discharged can be reduced, or any combination of these can be implemented.

With such a change in the processing conditions, the emission intensity I ₂₀ measured at 20 ° C. and the emission intensity I ₃₀ at 30 ° C. indicated by the black circles in FIG. 1 become the same as the value of the emission intensity I _37. It increases to the position of the white circle. Such processing conditions that can be changed are experimentally determined at the same concentration, and when measuring a plurality of biologically related substances whose concentrations are unknown, the processing conditions are changed according to the processing temperature to obtain the processing temperature. Regardless, it can be processed as if it was quantitatively determined at a constant temperature.

In other words, the control unit of the quantification system, when the biological substance is the same concentration, the actual measurement value (I ₂₀ or I ₃₀ ) at the processing temperature is the biological relation at the reference temperature (I ₃₇ ). as a measure of the reference substance (I ₃₇₎ substantially corresponding or substantially identical value (I ₃₇ '), sets the processing conditions. Here, “substantially corresponding” or “substantially identical” means a value preferably within ± 10%, more preferably a value within ± 5% with respect to a reference measurement value. After processing in this way, the bio-related substance can be quantified using a calibration curve as shown in FIG.

  In the embodiment of the present invention, the processing promoting tool is a solution mixing promoting means, and includes a tube 13 and a pump 20 (FIG. 2) described later. The processing region is a region including the tube 13 and the wells 100 to 110. The tube 13 can suck the solution containing the biological substance in the well 100, discharge the solution from the tube 13 to the wells 100 to 110, or move the solution to the beads 14 to 17 in the tube 13. it can. Processing is facilitated by such suction, discharge, and movement of the solution. Then, for example, the number of suction / discharge can be changed as shown in Table 1 according to the processing temperature. The “number of times of suction / discharge” defined a pair of suction and discharge as one time.

  The quantitative system according to the embodiment of the present invention includes a tube 13 as shown in FIG. The tube 13 has a target substance capturing bead 14 (first microparticle) and a negative control bead 15 (third microparticle) for detecting and quantifying a biologically relevant substance such as a protein contained in the specimen in the lumen. Microparticles), correction beads 16 (second microparticles), and light-shielding beads 17 (light-shielding members) disposed between the beads 14, 15, and 16. By arranging the light shielding bead 17 between the target substance capture bead 14, the negative control bead 15, and the correction bead 16, the measurement and quantification of the bead emitting fluorescence is not interfered with the light emission of the adjacent bead, It can be performed efficiently and accurately.

  The tube 13 shown in FIG. 2 is an embodiment of the tube including the above four types of beads, and is formed in a hollow cylindrical shape, and has an opening 18a formed by narrowing toward one end, An attachment portion 18b that can be attached to the mount portion 12 can also be provided at the other end. In this case, the mount 12 is provided with a nozzle (not shown) that communicates with the pump whose drive is controlled by the pump controller, and is mounted on the mount 12 so that the nozzle and the lumen of the tube 13 communicate with each other. Therefore, the tube 13 can suck liquid into the tube from the well 100 or the like and discharge the sucked liquid to the well 100 or the like outside the tube.

  Next, a quantitative system provided with the tube 13 described above will be described. The quantification system of the present invention refers to the measurement unit (light receiving unit 76) that measures signals emitted from the beads (microparticles) in the tube 13 in addition to the tube 13, and a calibration curve prepared in advance (FIG. 6). And a correction unit that corrects the detected signal data, and a calculation unit that quantifies the biological substance based on the corrected data. One form will be described below.

  The quantitative system 30 according to the embodiment of the present invention will be further described based on the block of FIG. The quantitative system 30 includes a control unit 32, a tube position control unit 34, a tube mount control unit 36, a temperature temperature measurement unit 40, a pumping control unit 42, a RAM 46, a ROM 48, a display panel 50, an operation interface 52, a signal processing circuit 54, and light emission. It has an intensity calculator 56, a corrected quantitative calculator 58, a timer (not shown), and the like, and is detachably equipped with a tube 13 for detecting and quantifying a target biological substance.

  The tube position control unit 34 includes XYZ axes orthogonal to each other, and controls the position of the nozzle by a stepping motor or a servo motor. The X axis and the Y axis are substantially parallel to the well plate and perpendicular to each other, and the Z axis is substantially perpendicular to the well plate. When moving the nozzle, for example, the tube (nozzle) is moved in two stages: movement on the X and Y axes, which are substantially parallel to the well plate, and movement on the Z axis, which is substantially perpendicular to the well plate. It is.

  Various control programs are stored in the ROM 48. A control program is expanded from the RO 48 to the RAM 46 in accordance with the operation mode selected by the user through the operation interface 52, and the central control unit 32 controls each part of the quantitative system 30 based on the control program expanded in the RAM 46. .

  The display panel 50 displays items that need to be presented to the user. For example, the processing conditions such as the number of pumping times during sample (specimen) pretreatment, the rate of flow rate during pumping, the amount of suction and discharge, the rate of movement of the tube 13 can be displayed on the display panel 50, and the user can It can be confirmed via the display. If it is desired to change various setting contents, it can be changed through operation of the operation interface 52.

  A timer unit (not shown) counts time according to the program read from the ROM 48. The time is counted when, for example, incubation is performed or pumping is performed, whereby each process is accurately executed. The temperature measurement unit 40 measures the temperature near the tube 13 (FIG. 2) and / or the wells 100 to 110 (FIGS. 2 and 5), preferably the temperature of the liquid contained in the tube 13 and / or the wells 100 to 110. To do.

  The tube mount control unit 36 attaches the tube 13 to the mount unit 12 and detaches the tube 13 from the mount unit 12. The tube mount control unit 36 is provided at a location some distance away from the well plate in which the wells in which the specimens are accommodated are arranged, so that contamination should not occur if liquid is scattered from the tube 13 when the tube 13 is replaced. Has been. The tube mount control unit 36 includes, for example, a holding unit that holds the tube 13 and a preparation unit that prepares another new tube 13. When the nozzle rises along the Z axis while the gripping part grips the tube 13, the tube 13 is detached from the nozzle. Next, the exposed nozzle moves on the X axis and the Y axis and moves above the new tube 13. In the preparation part, the posture of the new tube 13 is maintained with the mount part on the upper side and the tip part on the lower side, and the nozzle is lowered along the Z-axis so that the new tube 13 is mounted on the nozzle. Installed.

  The pumping control unit 42 includes the pump 20 and the pressure sensor 70, and controls the suction and discharge of the specimen performed through the nozzle and the tube 13 attached to the nozzle. The pump 20 includes, for example, a housing formed in a cylinder shape, a piston that is movably fitted in the housing, and a motor that drives the piston, and the inside of the housing communicates with an opening of a nozzle. The movement of the piston is controlled by, for example, a servo motor, and the drive of the servo motor is controlled by a drive control signal from the pumping control unit 42. When the piston is activated, liquid can be sucked or discharged through the nozzle opening.

  A pressure sensor 70 for detecting pressure is provided in the opening of the nozzle, and the pressure sensor 70 sends a pressure signal to the pumping control unit 42. The pumping control unit 42 monitors the pressure based on the pressure signal from the pressure sensor 70. With such a configuration, for example, when the tip of the tube 13 is immersed in the sample in the well, the pressure detected by the pumping control unit 42 exceeds a predetermined threshold value, and the servomotor is driven and controlled accordingly. A signal is sent out. The pressure sensor 70 always sends a pressure signal to the pumping control unit 42 at the time of suction and discharge of the specimen, so that the pumping control section 42 can control the drive of the servo motor with high accuracy, and the suction of the specimen. It is possible to monitor whether the pressure and the exhaust pressure are too low or too high, and thereby control whether suction and discharge are performed within a predetermined range.

  The signal processing circuit 54 processes the light reception signal from the light receiving unit 76 to form, for example, binarized light reception data. The light receiving unit 76 may include an image sensor such as a PMT (photomultiplier tube), a CCD image sensor, or a CMOS image sensor. FIG. 4 shows an example of a configuration when a PMT is used for the light receiving unit 76. As shown in FIG. 4, the light emission intensity measuring mechanism 79 includes a PMT 80, a light emitting POF (plastic optical fiber) 82a, a light receiving POF (plastic optical fiber) 82b, a ring member 84, and the like. Excitation light from a light source (not shown) is irradiated to each bead from the light emitting end of the light emitting POF 82a, and fluorescent light (signal) from each bead is guided from the light receiving end of the light receiving POF 82b to the light receiving POF 82b to the PMT 80. It is burned. The PMT 80, the POFs 82a and 82b, and the ring member 84 move along the longitudinal direction of the tube 13, and the PMT 80 outputs a light reception signal in response to the emission intensity of each bead.

  The signal processing circuit 54 includes a sampling circuit, an amplifier, an A / D conversion circuit, etc. (not shown), and amplifies and digitizes the light reception signal sent from the light receiving unit 76 to form light reception data. The formed light reception data is sent from the signal processing circuit 54 to the light emission intensity calculator 56. When the emission intensity of the measurement tube 13 is read from the opening side to the attachment side, for example, the received light data formed corresponding to the light from the negative control beads 15, the target substance capture beads 14, and the correction beads 16 is obtained. It is sent out from the light receiving unit 76.

  The light emission intensity calculation unit 56 reads a light emission intensity calculation program stored in the RAM 46, for example, and calculates the light emission intensity based on the received light data from the signal processing circuit 54 according to the read program. For example, the received light data corresponding to the light from the negative control beads 16, the target substance capturing beads 14, and the correction beads 16 arranged in the tube 13 is received from the signal processing circuit 54 and the light emission intensity is calculated. The luminescence intensity data calculated corresponding to the light from the correction bead 16 and the luminescence intensity data calculated corresponding to the light from the target substance capturing bead 14 are sent to the correction quantitative calculation unit 58 and from the negative control bead 16. The emission intensity data corresponding to the light is sent to the central control unit 32 and verified. In this test, for example, it is determined whether or not the emission intensity corresponding to the light from the negative control bead 16 is smaller than the emission intensity calculated corresponding to the light from the target substance capture bead 14. If it is determined that there is an abnormality, a warning is displayed on the display panel 50.

  The correction quantitative calculation unit 58 quantifies the target biological substance based on the emission intensity data corresponding to the light from the target substance capturing bead 14 and the emission intensity data corresponding to the light from the correction bead 16. When quantifying a biological substance, first, the target biological substance quantification procedure is as follows: (1) Calculate the deviation Δ of the measured value from the luminescence intensity predicted from the biological substance fixed in advance on the correction bead 16. . (2) The calculated deviation amount Δ is reflected in the emission intensity data of the target substance capturing bead 14. (3) The concentration corresponding to the recalculated emission intensity data is read out using a calibration curve. By executing the above procedure, the quantitative value of the biological substance captured by the target substance capture beads 14 can be made more accurate.

  Next, the operation of the quantitative system according to the embodiment of the present invention will be described with reference to an example in which an antigen is measured based on an antibody-antigen reaction. As shown in FIG. 5, the transparent tube 13 formed in a tubular shape includes a target substance capturing bead 14 in which an antibody against an antigen is fixed in advance, a correction bead 15, and a negative control bead 16. A light shielding bead 17 is arranged between the target substance capturing bead 14, the negative control bead 15, and the correction bead 16, and each bead 15 to 17 is arranged in a line along the extending direction of the tube 13.

  As shown in FIG. 5A, when the opening 18 a of the tube 13 is immersed in the well 100 containing the sample solution, and the sample solution in the well 100 is sucked into the tube 13, it is fixed to the target substance capturing bead 14. Antigens corresponding to the formed antibodies are bound and captured by the target substance capture beads 14. By repeating the suction and discharge of the specimen a predetermined number of times, the antigen can be more reliably bound to the antibody of the target substance capture bead 14. The predetermined number of times is changed by the control unit based on the processing temperature as shown in Table 1.

  As shown in FIG. 5B, after pumping the solution containing the biological substance, the cleaning liquid in another well 104 is sucked to wash the target substance-capturing beads 14. The components other than the antigen bound by the antigen-antibody reaction are removed by washing. As shown in FIG. 5 (c), after washing the target substance-capturing beads 14, the opening 18 a of the tube 13 is immersed in an enzyme-labeled antibody solution housed in another well 106, and the enzyme-labeled antibody solution is placed in the tube 13. Inhale. As shown in FIG. 5 (d), when pumping of the enzyme-labeled antibody solution is executed a predetermined number of times, the cleaning solution stored in another well 108 is pumped to flow out excess enzyme-labeled antibody solution, The target substance capture beads 14 to which the antigen and enzyme-labeled antibody are bound are washed. As shown in FIG. 5E, after the washing, the tube 13 is controlled to move to the well 110 containing the substrate solution for the luminescence reaction, and the detection process is started.

  The opening 18 a of the tube 13 is immersed in the well 110 in which the substrate solution for luminescence reaction is accommodated, and the substrate solution is sucked into the tube 13. The substrate solution is pumped for a set number of times based on the processing temperature and allowed to react sufficiently.

  The light emitting POF 82a, the light receiving POF 82b, and the PMT 80 (light receiving unit 76) are controlled to move so as to face each other through the beads, and the light beam from the light irradiation unit is received by the light receiving unit 76 through each bead of the tube 13. The Based on the light reception signal transmitted from the light receiving unit 76, the signal processing circuit 54 transmits light reception data based on the light reception signal from the light receiving unit 76 to the light emission intensity calculation unit 56. The light emission intensity calculation unit 56 calculates the light emission intensity based on the received light reception data and sends the light emission intensity data to the corrected quantitative calculation unit 58. The corrected quantitative calculation unit 58 corrects the emission intensity of the target substance capturing bead 14 based on the emission intensity data of the correction bead 15. In addition, when quantifying the target biological substance using a luminescence reaction, it is preferable to use light-shielding beads.

  In the above embodiment, one target substance capture bead, one correction bead, and one negative control bead are used. However, a plurality of beads may be used. By performing the quantification of the target substance or the like using the measurement average with a plurality of beads, it is possible to expect a highly reliable quantification by using the measurement average.

  When the second microparticle is used to correct the measurement value of the biological substance captured by the first microparticle, for example, the specimen is put in a tube, the microparticle and the specimen are brought into contact, and fixed to the second microparticle. When a signal (for example, fluorescence intensity) is measured for the bio-related substance, the emission intensity deviates from a value (value on the calibration curve) predicted in advance from the fixed amount of the bio-related substance of the second microparticle. is there. In such a case, a deviation amount Δ (delta) from the calibration curve is calculated, and this deviation amount Δ is reflected in the measurement data of the target substance-capturing beads to obtain a quantitative value of the biological substance. By quantifying the biological substance according to such a procedure, an error in quantifying the target biological substance can be reduced, and more accurate quantification can be performed.

  For example, as shown in FIG. 6, a predetermined amount (an amount corresponding to any position on the calibration curve) of a biological substance is fixed to the second microparticle, and the second microparticle relates to the second microparticle. The emission intensity is expected to be plotted at a position point P on the calibration curve with respect to the concentration A, for example. However, when the measured value of the emission intensity is plotted at the position Q deviated above the calibration curve, there is a deviation amount Δ of the emission intensity between the point P and the point Q. This deviation amount Δ can be used as a correction parameter for light emission intensity. The amount of deviation Δ is subtracted, for example, from the measured value of the emission intensity related to the first microparticle to which the substance capable of binding to the biological substance to be measured is fixed to correct the emission intensity. When the emission intensity of the first microparticle is β, the concentration B corresponding to the value γ obtained by subtracting the shift amount Δ from β is obtained as a more accurate quantitative value. By obtaining the concentration corresponding to the luminescence intensity subjected to the subtraction process via a calibration curve, the quantification of the biological substance bound to the first microparticle can be performed with higher accuracy. Further, when the emission intensity of the second microparticle with respect to the concentration A is plotted on the lower side of the calibration curve, the deviation amount Δ is added to the actual measurement value of the emission intensity related to the first microparticle to correct the emission intensity. To do. By obtaining the concentration corresponding to the luminescence intensity subjected to the addition process through a calibration curve, the quantification of the biological substance bound to the first microparticle can be performed with high accuracy. Note that the concept of correcting the light emission intensity can be applied not only to the light emission intensity described above but also to other measurement quantities such as absorbance, and is used by appropriately changing according to the configuration of the quantitative system. be able to.

  In addition, by using the first microparticle that captures the biological substance, the second microparticle in which a predetermined amount of the biological substance is fixed in advance, and the third microparticle for negative control, Even when the lot number, manufacturing number, type and model number of the quantification system, reagents used in quantification, and the like are different, it is possible to quantify the target biological substance more accurately while saving the time required for system calibration. Thus, a more convenient quantitative system can be provided by using the measurement tube and quantitative system of the present invention.

  Furthermore, in the measurement tube of the present invention and the quantitative system using this measurement tube, the maintenance and maintenance work can be saved, and a point-of-point for performing a diagnosis in the vicinity of the patient for quick diagnosis. Can respond sufficiently to the execution of care.

  In the embodiment of the present invention, the microparticle is formed in a substantially spherical shape, and the particle size is preferably 0.05 to 10.0 mm, more preferably 0.1 to 5.0 mm. Examples of the microparticle material include silicon nitride, silica, glass, magnetite, polystyrene, polyvinyl chloride, polyethylene, polypropylene, polycarbonate, nylon, polyacrylamide, dextran, amylose, agarose, natural and modified cellulose, activated carbon, and the like. can give.

  In the present invention, the “biologically related substance” is a substance to be quantified or detected contained in a sample, and is a microorganism (bacteria), virus, parasite, cell, nucleic acid, polysaccharide, protein (peptide, hormone, receptor). , Enzymes), antigens, antibodies, toxins, pathogens, small molecules and the like.

  In the embodiment of the present invention, the number of tubes is one. However, the present invention is not limited to this, and a plurality of, for example, 16 tubes and a plurality of corresponding wells are used to simultaneously quantify biological substances in a plurality of specimens. Can be carried out at room temperature. As a result, it is possible to make a plurality of specimens multi-flexible without performing temperature control. In the embodiment of the present invention, each calculation unit is provided independently of the control unit. However, these calculation units may be provided in the control unit.

  In the embodiment of the present invention, fluorescence was measured during quantification, but chemiluminescence can be used. Since chemiluminescence itself contains a chemical reaction, temperature dependence differs from that of fluorescence. Therefore, in the case of chemiluminescence, processing conditions different from the fluorescence processing conditions can be set. In the embodiment of the present invention, the antigen-antibody reaction is used. However, the present invention is not limited to this, and when the biological substance is a nucleic acid, hybridization of a nucleic acid fixed to the antibody can also be used.

  In the embodiment of the present invention, by verifying the correlation condition between the processing temperature and the fluorescence intensity, a control program that can automatically determine the suction and discharge conditions based on the room temperature during system operation is basically used. No quantitative system became possible. However, since there are upper and lower limits of the processing temperature that can be correlated, it is preferable to perform quantification when the processing temperature is in the temperature range between the upper limit and the lower limit.

  Temperature control is a major burden in establishing and manufacturing management of quantitative systems, and the ability to eliminate temperature control elements as in the present invention is a great merit for the low cost, small size, and ease of manufacture of quantitative systems. .

12 Mount part 13 Tube 14 Target substance capture bead (first microparticle)
15 Negative control beads (third microparticle)
16 Correction beads (second microparticle)
17 Shading beads 30 Quantitative system 32 Control unit 34 Tube position control unit 36 Light receiving unit 46 RAM
48 ROM
54 Signal Processing Circuit 56 Emission Intensity Calculation Unit 58 Correction Quantitative Calculation Unit

Claims (17)

A system for quantifying biological materials,
A treatment region for performing a predetermined treatment for quantifying the biological substance in the solution; a treatment accelerator for promoting the treatment in the treatment region; and a temperature measurement for measuring the treatment temperature of the treatment region during the treatment. A controller that sets a processing condition other than the processing temperature based on a comparison between the processing temperature and a predetermined reference temperature, and operates the processing accelerator based on the set processing condition; A measurement unit that measures the biological substance in order to quantify the biological substance after activating the processing accelerator;
In the control unit, when the biological substance is set to the same concentration, the measurement value at the processing temperature substantially corresponds to or substantially the same as the reference measurement value of the biological substance at the reference temperature. A system for changing the processing conditions so that   2. The system for quantifying a biological substance according to claim 1, wherein the processing condition includes a time during which the processing accelerator performs the processing, a speed at which the processing accelerator performs the processing, and the processing accelerator mixes the solution. The system which is any one or any combination of the number of times the processing facilitating tool is mixed, and the amount of liquid with which the processing accelerator is mixed.   3. The system for quantifying a biological substance according to claim 1, wherein the processing region accommodates one or a plurality of tubes capable of sucking and discharging the solution, and the solution movably between the tubes. System with one or more wells. The system for quantifying a biological substance according to claim 3,
The processing accelerator includes a pump for sucking and discharging the solution to the tube.   The system for quantifying a biological substance according to any one of claims 1 to 4, wherein the reference temperature is room temperature or body temperature.   The system for quantifying a biological substance according to claim 1, wherein the measurement unit measures fluorescence of a substance combined with the biological substance.   The system according to any one of claims 1 to 6, wherein the treatment includes an antigen-antibody reaction or nucleic acid hybridization. In the system for quantifying a biological substance according to any one of claims 1 to 7,
The control unit sets the processing condition when the processing temperature measured by the temperature measuring unit is included in a predetermined temperature range including the reference temperature.   The system according to claim 8, wherein the predetermined temperature range is a range from about 20 ° C to about 30 ° C, or a range from about 20 ° C to about 37 ° C.   The system for quantifying a biological substance according to claim 8, wherein the predetermined temperature range is 15 ° C ± 5 ° C, 20 ° C ± 5 ° C, 25 ° C ± 5 ° C, or 30 ° C ± 5 ° C.   11. The system for quantifying a biological substance according to claim 1, wherein the reaction region includes a first microparticle to which a substance capable of binding to a biological substance to be measured is fixed. A system in which the second microparticles in which the biological substance is fixed in a predetermined amount and the third microparticles for negative control in which the biological substance is processed so as not to be bound are arranged in a line. The system for quantifying a biological substance according to claim 11,
The system, wherein the first microparticle is a microparticle for correcting measurement data of the biological substance. The system for quantifying a biological substance according to claim 12 or 13,
The system, wherein the second microparticle is a plurality of microparticles to which different amounts of biological substances are immobilized. The system for quantifying a biological substance according to claim 13,
The system, wherein the plurality of microparticles are microparticles for creating a calibration curve. In the quantification system of the living body relevant substance according to any one of claims 11 to 14,
A system further comprising a light-shielding member between the first to third microparticles. In the quantification system of the living body relevant substance according to any one of claims 11 to 14,
The measurement unit detects a signal emitted from the microparticle in the tube,
A system in which the control unit corrects data of the detected signal with reference to a calibration curve prepared in advance, and quantifies a biological substance based on the corrected data. In the quantification system of the living body relevant substance according to any one of claims 11 to 14,
The measurement unit detects a signal emitted from the microparticle in the tube,
The system, wherein the control unit creates a calibration curve based on the detected signal, and quantifies a biological substance with reference to the created calibration curve.