JP2004309233A - Chemical analysis system - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a chemical analysis system which derives and analyzes chemical substances being objects to be inspected from a sample in a short period of time. <P>SOLUTION: The chemical analysis system is provided with a housing section of a structure having a sample supply section by which the sample to be inspected is supplied; a reagent holding section which holds reagents; a mixing section into which the sample from the sample supply section and the reagent from the reagent holding section are supplied; and a trapping section into which mixed fluid is introduced and by which a specific chemical substance in the sample is trapped. The sample supply section includes a detecting device into which indicators flowing downward together with the sample in the structure are supplied and which is installed outside the structure and detects the indicators flowing into the structure. <P>COPYRIGHT: (C)2005,JPO&NCIPI

Description

[0001]
TECHNICAL FIELD OF THE INVENTION
The present invention relates to a chemical analyzer for analyzing components in a sample.
[0002]
[Prior art]
WO 00/78455 describes a microstructure and method for inspection using amplification. In this apparatus, a DNA mixture is passed through a glass filter using a method for purifying and separating a nucleic acid mixture, and then is passed through a washing solution and an eluent to collect only DNA. The glass filter is provided on a rotatable structure, and reagents such as a washing liquid and an eluent are held in respective reagent reservoirs in the same structure. Each reagent flows by the centrifugal force generated by the rotation of the structure, and the reagent passes through the glass filter by opening a valve provided in a fine flow path connecting each reagent reservoir and the glass filter.
[0003]
As a chemical analyzer for extracting and analyzing a specific chemical substance such as a nucleic acid from a sample containing a plurality of chemical substances, JP-A-2001-527220 describes an integrated fluid operation cartridge. In this apparatus, reagents such as a lysis solution, a washing solution, and an eluent, and a capture component for capturing nucleic acids are provided inside the integrated cartridge, and after a sample containing nucleic acids is injected into the cartridge, the sample and the eluent are mixed. And pass the capture component, further pass the wash component through the capture component, further pass the eluent through the capture component, contact the eluate after passing through the capture component with the PCR reagent and into the reaction chamber And shedding.
[0004]
JP-T-2001-502793 discloses an apparatus and a method for chemical analysis. In this device, a chamber, a flow path, a reservoir, and an analysis cell are provided inside a disk-shaped member, a blood sample is injected into a centrifugal chamber, blood cells and serum are centrifuged, and the serum is coated with beads coated with a reagent. Only the serum is flowed into the reaction chamber, the solution for washing is then flowed into the reaction chamber, the elution solution is further flowed into the reaction chamber, and the elution solution is moved from the reaction chamber to a plurality of analysis cells. There are a plurality of analysis cells, which are dispensed along a certain time series. The inlet of the analysis cell is on the rotation center side.
[Patent Document 1]
WO 00/78455
[Patent Document 2]
JP 2001-527220 A
[Patent Document 3]
JP 2001-502793 A
[0005]
[Problems to be solved by the invention]
In a chemical analyzer that extracts and analyzes a specific chemical substance from a sample containing the chemical substance, reducing the time required for the analysis is an important issue. However, the prior art does not describe a method of monitoring in the course of analysis, a method of responding to the result of monitoring, and an apparatus that enables the method.
[0006]
An analysis often includes a plurality of steps such as separation, mixing, and washing, and monitoring each step and immediately moving to the next step at the end of the step leads to a reduction in operation time. In addition, monitoring for each process enables speed control, acceleration, etc. to be performed in response to equipment failures even if equipment failures occur, minimizing damage such as detection failures. it can.
[0007]
Therefore, an object of the present invention is to solve the above-mentioned problems and to provide a chemical analyzer for extracting and analyzing a chemical substance to be inspected from a sample in a short time.
[0008]
[Means for Solving the Problems]
The above problem can be solved by supplying a chemical analysis device provided with a detection device for monitoring an intermediate process in a process of analyzing a specific substance in a sample.
(1) a sample supply section to which a sample containing a specific chemical substance and a label are supplied, a reagent holding section to hold a reagent, the sample and the label to be supplied from the sample supply section, and A mixing unit to which a reagent is supplied, and the mixed fluid is introduced, and a capturing unit that captures the chemical substance.A storage unit of a structure, and a storage unit that is installed outside the structure and is provided inside the structure. And a detecting device for detecting the marker that is located.
[0009]
Further, it is preferable to perform control so as to change the contents of the steps required for the analysis according to the detection result of the detection device.
[0010]
For example, in (1), the accommodating portion includes a mechanism for attaching and detaching the structure, and a driving device for driving the structure to rotate, and the driving rotation speed is determined based on a detection value of the marker in the detection device. Control to change
Further, the end of the small steps constituting all the steps in the analysis can be determined based on the detection result of the label by the detection device. Further, it is possible to have a means for judging that the flow of the sample or the reagent is stagnant, based on the detection result of the label of the detection device, and promoting the flow.
[0011]
When the detected value is low, it can be determined that the flow of the sample is not sufficient, and the rotation speed can be controlled to be higher than before. Alternatively, when the detected value is sufficient, it is determined that the desired flow of the sample is sufficient, and the control can be performed so as to end the process and shift to the next process.
[0012]
The label may include at least one of a luminous body, fine particles smaller than 100 nanometers, a dye, a magnetic substance, an odor molecule, and an organic solvent, or a plurality of them.
[0013]
Thus, it is possible to provide a chemical analyzer for extracting and analyzing a chemical substance to be inspected from a sample in a short time.
[0014]
By introducing a mixture of a sample and a detection agent into the chemical analyzer and detecting the detection agent, the state of the process can be monitored.By detecting at the end of each step required for analysis, The processing time of the process can be shortened, and further, by detecting a malfunction of the operation, follow-up becomes possible, and accuracy can be improved.
[0015]
Investigations on the use of the label revealed that there were JP-A-6-94676, JP-A-11-230968, and the like. It does not disclose a form of detection by flowing down.
[0016]
BEST MODE FOR CARRYING OUT THE INVENTION
An embodiment in which the chemical analyzer according to the present invention is applied to a gene analyzer will be described with reference to FIGS. Note that the present invention is not limited to the contents disclosed in the specification, and does not exclude a change based on a well-known technology or a technology that will become a well-known technology in the future.
(Example 1)
Hereinafter, a case where the optical device is not provided on the holding disk will be described.
[0017]
FIG. 1 is an overall configuration diagram of a chemical analyzer according to the present invention. The gene analyzer 1 includes a motor 11, a holding disk 12 rotatably supported by the motor 11, a plurality of fan-shaped analysis disks 2 positioned by protrusions 121 on the holding disk 12, and a holding disk. A positioning projection 17 for determining the position of the liquid, a position detector 16, a drilling machine 13 having a drilling machine for controlling the flow of liquid and an air introducing device for pushing out air, Optical device 14 is provided. The optical device 14 includes an optical filter for detection. The optical device 14 detects a plurality of fluorescence spectra by switching optical filters.
[0018]
FIG. 2 is a sectional view of a set of the analysis disk 2. The analysis disk 2 is configured by joining a lower cover 40 (FIG. 3), a flow path 30 (FIG. 4), and an upper cover 20 (FIG. 5). The upper cover 20 includes a sample inlet 1020, a plurality of reagent inlets 1010, 1011, 1012, 1013, 1014, 1015, 1016, 1017 and a plurality of vents 1021, 1022, 1023, 1024, 1025, 1026, 1027, 1028. , 1029, 1030, a serum separation section observation window 1040 for observing the serum separation section 1050 of the flow path section 30, a capillary observation window 1041 for observing the capillary 1054, and a mixing vessel observation window for observing the mixing vessel 1057. 1042, a waste liquid container observation window 1043 for observing the waste liquid collection container 1063, and a detection container observation window 1044 for observing the detection container 1068. The lower cover 40 has a positioning hole 460. The analysis disk 2 is positioned by fitting the positioning holes 460 to the protrusions 121 of the holding disk 12.
[0019]
FIG. 4 shows a configuration diagram of the flow path unit 30. The embodiment of the flow path unit shown in FIG. 4 constitutes a flow path for extracting nucleic acid contained in virus in serum and analyzing the serum after separating serum from whole blood.
The operation of extracting and analyzing viral nucleic acids when whole blood is used as a sample will be described below. The flow of the extraction and analysis operation is shown in FIG. 6, FIG. 7, and FIG.
[0020]
The operator dispenses a reagent from the upper cover 20 of the analysis disk 2 through each reagent inlet to each reagent container, and covers the reagent container. After injecting the reagent into a required number of analysis disks according to the number of analysis, the analysis disks are mounted on the holding disk 12.
[0021]
Next, two kinds of fluorescent beads as a detection agent, for example, a fluorescent bead having an emission spectrum of 580 nm, a diameter of 20 nm, a specific gravity of 1.05, an emission spectrum of 440 nm, and a diameter are added to whole blood collected by a vacuum blood collection tube or the like. Fluorescent beads having a specific gravity of 15 μm and a specific gravity of 1.05 are mixed and injected into the sample container 310 from the sample inlet 210 (FIG. 9). The injected whole blood 1110 is shown in FIG. After the whole blood is injected, the holding disk 12 is rotated by the motor 11. The whole blood injected into the sample container 310 flows to the outer peripheral side by the action of the centrifugal force generated by the rotation of the holding disk 12, and flows into the blood cell separation unit 1050. At this time, the whole blood exceeding the capacity of the serum separation unit 1050 flows through the whole blood discarding channel 1051 and enters the whole blood discarding container 1052.
[0022]
As the number of rotations of the holding disk 12 increases, blood cell components (specific gravity of red blood cells 1.095, white blood cells 1.063 to 1.085) and serum (specific gravity of platelets 1.032, plasma 1.025 to 1.029) are increased. Separates gradually due to the difference in specific gravity. At this time, the beads of φ20 nm do not precipitate under the influence of Brownian motion and most of the beads remain on the serum side 1111, but the beads of φ15 μm move to the blood cell holding container 1053 together with the blood cells 1112 (FIG. 10). The optical device 14 is moved onto the serum separation unit observation window 1040. If the optical device 14 cannot detect the fluorescence of the beads of φ15 μm from the serum separation section observation window 1040, it is determined that the separation of serum and blood cells in whole blood has been completed, and the process proceeds to the next step.
[0023]
In the present embodiment, a sample inlet 210 to which a blood sample containing blood cells and serum and a marker flowing down in the structure together with the sample are supplied, and an area mainly containing the blood cells separated downstream of the sample inlet 210 And a serum separation unit 1050 having a region mainly storing the separated serum, a holding disk 12 which is a storage unit for storing the analysis disk 2 which is a structure, and a motor 11 which is a driving device for rotating the disk. And a detection device installed outside the analysis disk 2 for detecting a label flowing down the flow path in the analysis disk 2; and a rotation device for separating serum and blood cells in the sample. When the reaction of the label detected by the detection device from the analysis disk 2 after performing the detection is equal to or less than a predetermined value, control is performed to perform rotation for separating serum and blood cells again. .
[0024]
On the other hand, for example, as described above, when the reaction of the marker detected by the detection device is larger than a predetermined value, the separation step ends. Then, the process proceeds to the next step.
Further, at the time of the re-rotation, the structure is controlled so that the structure is re-rotated at a rotation speed higher than the rotation speed of the rotation before the detection. Further, for example, when the separation of serum and blood cells is not completed within a predetermined time, the rotation speed of the motor 11 is further increased.
[0025]
When the motor 11 is stopped and the holding disk 12 is stopped, the separated serum 1111 flows through the capillary 1054 by capillary action (FIG. 11). The optical device 14 is moved onto the capillary detection window 1041, and when the fluorescence of the φ20 nanometer beads is detected from the capillary detection window by the optical device 14, it is determined that the serum has flowed to the capillary outlet 1055, and the next step is performed. Move on to In this step, if the fluorescence of the φ20 nanometer fluorescent beads cannot be detected within a predetermined time, the sample injection is performed by the air introducing device 13 until the fluorescence of the φ20 nanometer fluorescent beads is detected from the capillary detection window 1041. From the inlet 210, the serum remaining in the middle of the capillary is pushed out to the capillary outlet 1055 by applying air pressure.
[0026]
In addition, the sample inlet 210 to which the blood sample containing blood cells and serum and the label flowing down in the structure together with the sample are supplied, and the blood cells separated downstream of the sample inlet 210 are mainly stored. A serum separation section 1050 having a region and a region mainly containing the separated serum, a capillary tube 1054 through which a fluid containing the separated serum flows, and a flow path communicating with a reagent container such as a lysis solution holding container 311 are provided. The analysis device includes a holding disk 12 that is a storage unit that stores the analysis disk 2 that is a structure including the mixing unit 1056 and the mixing container 1057 that are connected to each other, and a motor 11 that is a driving device that rotates the disk. A detection device installed outside the disk 2 for detecting a marker flowing down the flow path in the analysis disk 2; If response is not more than a predetermined value, characterized in that the fluid to promote flow down into the fluid in the capillary tube 1054 containing serum is controlled to be supplied from the upstream side of the serum separation unit 1050. In the above example, air is used as the fluid, but other liquids may be used. Then, as described above, when the reaction of the detected marker is larger than the predetermined value, the process of flowing the fluid into the capillary is completed. It is possible to move to the next step.
[0027]
After confirming that the serum has filled the capillaries, proceed to the next step. The cover of the solution vent 1024 is pierced by the piercing machine 13 and the motor 11 is rotated. The serum held in the serum holding container and the lysate held in the lysate holding container 311 simultaneously pass through the lysate outflow channel 312 and the mixing unit 1056 and flow into the mixed solution holding container. When the serum and the lysis solution flow into the mixing container 1057, the mixed solution is once mixed because of the flow channel structure (the mixed solution return flow channel 1059) that returns the mixed solution 1113 of the serum and the lysis solution to the inner peripheral side once. It is held in the container 1057 (FIG. 12), and the serum and the lysis solution are mixed in the mixing container 1057. The mixing container 1057 has a two-stage structure as shown in FIG. There is a mixing vessel detection window 1042 just above the upper stage 1058 of the reaction vessel. The optical device 14 is moved onto the mixing container detection window 1042. The mixed state of the lysate and the serum is determined from the distribution of fluorescent beads having a diameter of 20 nm contained in the serum. The light emission of the fluorescent beads detected from the mixing container detection window 1042 of the rotating analysis disk is detected by the optical device 14, and the distribution of the φ20 nanometer fluorescent beads in the reaction container is examined. The mixed state is determined based on the magnitude of the difference between the distributions. After confirming that the components are sufficiently mixed, the process proceeds to the next step.
[0028]
When the motor 11 is stopped and the additional liquid container vent 1025 is pierced by the piercing machine 13 and the motor 11 is rotated, the additional liquid flows into the mixing container 1057, the amount of the mixed liquid in the mixing container 1057 increases, and the mixed liquid return flow path When the mixed liquid 1113 exceeds the innermost peripheral side of 1059, the mixed liquid 1113 flows out of the mixed liquid return flow path 1059 to the nucleic acid binding member 301 through the mixed liquid outflow flow path 1060. Use the same solution as the dissolving solution for the additional solution.
[0029]
The lysing solution functions to dissolve the membrane from the virus or bacteria in the serum to elute the nucleic acid, and further promotes the adsorption of the nucleic acid to the nucleic acid binding member 301. As such a reagent, guanidine hydrochloride may be used for elution and adsorption of DNA, and guanidine thiocyanate may be used for RNA, and a porous material such as quartz or glass or a fiber filter may be used as a nucleic acid binding member. Further, since a nucleic acid binding member 301 having a minimum collection diameter larger than φ20 nanometers is used, the fluorescent beads having a diameter of φ20 nanometers pass without being physically collected by the nucleic acid binding member.
[0030]
When the mixture of the lysis solution and the serum passes through the nucleic acid binding member in this way, the nucleic acid is adsorbed on the nucleic acid binding member 301, and the liquid passes through the waste liquid container return flow path 1061, the waste liquid container flow path 1062, and the waste liquid collection vessel 1063. (FIG. 14). The optical device 14 is moved to the height 1114 of the water surface when the total amount of the mixture of the lysate and the serum has flowed in, and the fluorescent mixture of 20 nanometers is detected from the waste liquid collection container window 1043 by the nucleic acid. Check that it has passed through the coupling member. If the fluorescence of the fluorescent beads of 20 nm is not detected, the rotation speed of the motor 11 is further increased.
[0031]
The motor 11 is stopped, and the first cleaning liquid vent 1026 is pierced by the piercing machine 13. When the motor 11 is rotated, the first cleaning liquid passes through the nucleic acid binding member 301 from the first cleaning liquid holding container 1064 via the first cleaning liquid outflow channel 1065, and reaches the waste liquid collecting container 1063 (FIG. 15). The optical device 14 is moved to the height 1115 of the water surface when the total amount of the lysis solution and serum that have flowed in previously and the first flow of newly flowing cleaning solution has flowed in, and the liquid waste collection container window 1043 is moved to 20 nanometers. By detecting the fluorescent beads, it is confirmed that the first washing solution has passed through the nucleic acid binding member.
[0032]
In addition, the sample inlet 210 to which the blood sample containing blood cells and serum and the label flowing down in the structure together with the sample are supplied, and the blood cells separated downstream of the sample inlet 210 are mainly stored. A serum separation section 1050 having a region and a region mainly containing the separated serum, a capillary tube 1054 through which a fluid containing the separated serum flows, and a flow path communicating with a reagent container such as a lysis solution holding container 311 are provided. A nucleic acid capturing unit including a mixing unit 1056 and a mixing container 1057 to be connected, a nucleic acid binding member 301 for capturing a specific nucleic acid in the mixed fluid into which the mixed fluid has been introduced via the mixing unit, and a nucleic acid capturing unit for the nucleic acid capturing unit A holding disk 12 which is a storage unit for storing the analysis disk 2 which is a structure including a waste liquid recovery container 1063 in which a mixed fluid passing through the storage unit is stored; A motor 11 serving as a driving device for rotating, and a detection device installed outside the analysis disk 2 for detecting a marker flowing down a flow path in the analysis disk 2. When the reaction of the sign detected by the detection device from the structure after the rotation for introducing the fluid into the waste liquid collecting container 1063 is equal to or less than a predetermined value, the rotation for introducing the mixed fluid into the waste liquid collecting container 1063 again is performed. It can be characterized by being controlled.
[0033]
Further, at the time of this re-rotation, control is performed so that the structure is re-rotated at a rotation speed higher than the rotation speed of the rotation before the detection. For example, when the fluorescence of the fluorescent beads of 20 nm is not detected, the rotation speed of the motor 11 is further increased. As the first cleaning solution, a solution having the same components as the solution and having substantially the same concentration is used.
[0034]
The motor 11 is stopped, and the second cleaning liquid vent 1027 is pierced by the piercing machine 13. When the motor 11 is rotated, the second cleaning liquid passes through the nucleic acid binding member 301 via the second cleaning liquid holding container 1066 and the second cleaning liquid outflow channel 1067, and reaches the waste liquid recovery container 1063 (FIG. 16). The optical device 14 is moved to the height 1116 of the water surface at the time when the total amount of the mixed solution of the lysis solution and serum, the first washing solution, and the newly washed second washing solution that have flowed before flows into the waste liquid collecting container window 1043. , 20 nanometer fluorescent beads, it is confirmed that the second washing solution has passed through the nucleic acid binding member 301. If the fluorescence of the fluorescent beads of 20 nm is not detected, the rotation speed of the motor 11 is further increased. A solution containing ethanol at a high concentration is used as the second cleaning solution.
[0035]
The motor 11 is stopped, and the lid of the detection container vent 1028 is pierced by the piercing machine 13. When the motor 11 is rotated, the channel cleaning liquid previously sealed in the detection container 1068 passes through the waste liquid container return flow channel 1061 and the waste liquid container flow channel 1062, and flows into the waste liquid recovery container 1063. Water or a solution having a low salt concentration is used as the channel cleaning liquid.
[0036]
The motor 11 is stopped, and the lid of the sealing liquid vent 1029 is pierced by the piercing machine 13. When the motor 11 is rotated, the sealing liquid flows into the waste liquid container return flow path 1061, but stops in the middle of the flow path due to its viscosity and the return structure of the waste liquid container return flow path 1061. The stopped sealing liquid 1117 is shown in FIG. As the sealing liquid, a gel adjusted so as to have a specific gravity higher than that of the eluent used in the subsequent step is used.
[0037]
When the motor 11 is stopped and the lid of the eluent vent 1030 is pierced by the punch 13 and the motor 11 is rotated, the eluent passes through the eluent holding container 1069, the eluent outflow channel 1070, and the nucleic acid binding site 301. Then, it flows into the detection container 1068 (FIG. 18). The nucleic acid adsorbed on the nucleic acid binding site 301 elutes into the eluent. As the eluent, water or a solution having a low salt concentration is used.
[0038]
When the motor 11 is stopped and the detection liquid vent 1031 is pierced by the piercing machine 13 and the motor 11 is rotated, the detection liquid flows into the detection container 1068. The detection solution is a reagent for amplifying and detecting a nucleic acid, and contains a primer, a nucleic acid synthase, deoxyribotriphosphate, a buffer, a metal ion, and the like necessary for amplification. Depending on the method of amplification or detection, it may contain a fluorescent dye, a primer modified with a fluorescent dye or biotin, or deoxyribotriphosphate modified with a fluorescent dye.
[0039]
The motor 11 is stopped, and the optical device 14 heats the detection container 1068 to a temperature suitable for the amplification method. For example, in the case of the constant temperature amplification method and the LAMP method, 55-65 ° C. is desirable.
After the amplification is completed, the optical device 14 is moved onto the detection container observation window 1044, and the amount of light emission is measured by the optical device 14.
[0040]
FIG. 19 shows the relationship between the monitoring process in the middle of the entire process and the method of responding to the monitoring result.
[0041]
At the time of perforation, heating, and detection, it is necessary to stop the holding disk 12 at a predetermined position. As shown in FIG. 1, a positioning projection 17 is provided on the holding disk 12, and the rotation position of the holding disk is detected by a position detector 16, and the rotation of the motor 11, the rotation and vertical movement of the punch 13, The rotation, irradiation, and detection of the device 14 are controlled.
[0042]
For example, FIG. 20 shows the operation timing of the drilling machine 13. The holding disk 12 lowers the rotation speed after the end of the flow of whole blood or each reagent, and maintains the low-speed rotation for positioning. When the position detector 16 detects the positioning projection 17, the holding disk 12 is stopped, and the punch 13 is lowered to make a hole in the lid of the vent hole of each reagent storage container, and then rise again. The post-perforation holding disk 12 rotates at such a low speed that the reagent does not flow out of the reagent storage container after the perforation, and rotates at the next analysis disk position, that is, 60 degrees when six analysis disks are mounted. Stop and repeat the same perforation operation. The location of the analysis disk may be determined by, for example, irradiating light with the optical device 14 and examining reflected light from the analysis disk. After all the analysis disks have been pierced, the holding disk rotates at high speed to allow the reagent to flow.
[0043]
According to the present invention, since the process for purifying and detecting the nucleic acid of the virus contained in the sample is constantly monitored, the end of each process can be immediately sensed. , The processing time of all the steps can be reduced.
Furthermore, according to the present invention, a high-sensitivity detector is not required for monitoring because a detection agent which is easy to detect is added instead of detecting a component contained in the sample in advance. In addition, since a fixed amount of the detection agent is added, the variation is smaller than in the case of detecting a component in a sample.
[0044]
(Example 2)
Hereinafter, a case where the holding disk includes an optical device will be described.
Basically, an embodiment in which the chemical analyzer according to the present invention is applied to a gene analyzer will be described with reference to FIGS.
[0045]
Basically, it can have the configuration shown in FIG.
FIG. 1 is an overall configuration diagram of the gene analyzer according to the present invention. Note that the light emitter 14 includes an optical filter for detection. The light emitting device 14 is characterized in that a plurality of excitation lights are emitted by switching an optical filter. Corresponding to this, the form and process of the first embodiment are provided.
[0046]
For example, the holding disk 12 includes a plurality of light receivers for detecting light emission from fluorescent beads or fluorescent dyes. Each of the light receivers has an optical filter suitable for the light to be received. FIG. 21 shows the configuration of the holding disk 12. On the holding disk, there are provided a serum separating photodetector 2010, a capillary photodetector 2011, a mixing container photodetector 2012, a waste liquid photodetector 2013, a detection container photodetector 2014, and a projection 121.
[0047]
Note that even if one kind of fluorescent beads, for example, fluorescent beads having an emission spectrum of 580 nm, a diameter of 20 nm, and a specific gravity of 1.05 can be used, the operations shown in FIGS. That is, as the rotation speed of the holding disk 12 increases, the blood cell component and the serum gradually separate due to the difference in specific gravity. At this time, the beads of φ20 nm do not precipitate due to the Brownian motion, and most of the beads remain on the serum side. The light emitter 14 is moved above the serum separation part observation window 1040, and is irradiated with light having a spectrum near 540 nm, which is the absorption wavelength of hemoglobin. At this time, if serum separation is insufficient, the absorption spectrum of the serum part shifts to the absorption spectrum of hemoglobin because blood cells are mixed in the serum part. When blood cell contamination is detected in the serum portion, the holding disk 12 is rotated by the motor 11 to perform serum separation again. At this time, the rotation speed is further increased. If it becomes impossible to detect the absorption of blood cells from the serum separating unit photodetector 2010, it is determined that the separation of serum and blood cells in whole blood has been completed, and the process proceeds to the next step.
When the motor 11 is stopped and the disc is stopped, the separated serum 1111 flows through the capillary 1054 by capillary action (FIG. 11). The light emitter 14 is moved above the capillary detection window 1041 and irradiated with excitation light of fluorescent beads having a diameter of 20 nm. At the stage where the fluorescence of the beads having a diameter of 20 nanometers is detected from the capillary light receiver 2011, it is determined that the serum has flowed to the capillary outlet 1055, and the process proceeds to the next step. In this step, if the fluorescence of the fluorescent beads with a diameter of 20 nanometers cannot be detected within a predetermined time, the air introducer 13 uses the capillary light receiver 2011 until the fluorescence of the fluorescent beads with a diameter of 20 nanometers is detected. The serum remaining in the middle of the capillary is pushed out to the capillary outlet 1055 from the sample inlet 210 by applying air pressure.
[0048]
After confirming that the serum has filled the capillaries, the process proceeds to the next step as in Example 1. The cover of the solution vent is pierced by the piercing machine 13 and the motor 11 is rotated. The serum held in the serum holding container 1050 and the lysate held in the lysate holding container 311 simultaneously pass through the lysate outflow channel 312 and the mixing unit 1056, and flow into the mixed solution container 1057. When the serum and the lysis solution flow into the mixing container 1057, the mixed solution is once mixed because of the flow channel structure (the mixed solution return flow channel 1059) that returns the mixed solution 1113 of the serum and the lysis solution to the inner peripheral side once. It is held in the container 380 (FIG. 12), and the serum and the lysis solution are mixed in the mixing container 1057. The reaction vessel has a two-stage structure as shown in FIG. There is a mixing vessel detection window 1042 just above the upper stage 1058 of the reaction vessel. The light emitter 14 is moved above the mixing container detection window 1042 and irradiated with excitation light of fluorescent beads having a diameter of 20 nm. The mixed state of the lysate and the serum is determined from the distribution of fluorescent beads having a diameter of 20 nanometers contained in the serum. The light emission of the fluorescent beads detected from the reaction container 1057 of the rotating analysis disk is detected by the reaction container light receiver 2012, and the distribution of the fluorescent beads in the reaction container is examined. The mixed state is determined based on the magnitude of the difference between the distributions. After confirming that the components are sufficiently mixed, the process proceeds to the next step in the same manner as in Example 1.
[0049]
When the mixture of the lysis solution and the serum passes through the nucleic acid binding member, the nucleic acid is adsorbed to the nucleic acid binding member, and the liquid flows into the waste liquid container return flow path 1061, the waste liquid container flow path 1062, and the waste liquid recovery container 1063 (FIG. 14). The light emitting device 14 is moved to a height 1114 of the water surface when the total amount of the mixed solution of the lysing solution and the serum has flowed in, and is irradiated with excitation light of fluorescent beads having a diameter of 20 nanometers. By detecting the nanometer fluorescent beads, it is confirmed that the mixture has passed through the nucleic acid binding member. If the fluorescence of the fluorescent beads of 20 nm is not detected, the rotation speed of the motor 11 is further increased.
[0050]
The motor 11 is stopped, and the first cleaning liquid vent 1026 is pierced by the piercing machine 13. When the motor 11 is rotated, the first cleaning liquid passes through the nucleic acid binding member 301 from the first cleaning liquid holding container 1064 via the first cleaning liquid outflow channel 1065, and reaches the waste liquid collecting container 1063 (FIG. 15). The light-emitting device 14 is moved to the height 1115 of the water surface when the total amount of the mixture of the lysis solution and the serum that has flowed in previously and the first washing solution that has newly flowed in, and the excitation light of the fluorescent beads of φ20 nanometers is moved. By irradiating and detecting 20 nanometer fluorescent beads from the waste liquid collection container light receiver 2013, it is confirmed that the first cleaning liquid has passed through the nucleic acid binding member. If the fluorescence of the fluorescent beads of 20 nm is not detected, the rotation speed of the motor 11 is further increased. As the first cleaning solution, a solution having the same components as the solution and having substantially the same concentration is used.
[0051]
The motor 11 is stopped, and the second cleaning liquid vent 1027 is pierced by the piercing machine 13. When the motor 11 is rotated, the second cleaning liquid passes through the nucleic acid binding member 301 via the second cleaning liquid holding container 1066 and the cleaning liquid outflow channel 1067, and reaches the waste liquid collecting container 1063 (FIG. 16). The light emitting device 14 is moved to a height 1116 of the water surface when the total amount of the mixture of the lysis solution and serum, the first washing solution, and the newly washed second washing solution which have flowed before, and the fluorescent beads of φ20 nanometers are moved. Irradiating the nucleic acid binding member by detecting the fluorescent beads of 20 nanometers from the waste liquid collecting vessel light receiver 2013 by irradiating the excitation light of the above. If the fluorescence of the fluorescent beads of 20 nm is not detected, the rotation speed of the motor 11 is further increased. A solution containing ethanol at a high concentration is used as the second cleaning solution.
[0052]
The motor 11 is stopped, and the detection container vent 1028 is pierced by the piercing machine 13. When the motor 11 is rotated, the channel cleaning liquid previously sealed in the detection container 1068 passes through the waste liquid container return flow channel 1061 and the waste liquid container flow channel 1062, and flows into the waste liquid recovery container 1063.
The motor 11 is stopped, and the sealing liquid vent 1029 is pierced by the piercing machine 13. When the motor 11 is rotated, the sealing liquid flows into the waste liquid container return flow path 1061, but stops in the middle of the flow path due to its viscosity and the return structure of the waste liquid container return flow path 1061. The stopped sealing liquid 1117 is shown in FIG. As the sealing liquid, a gel adjusted so as to have a specific gravity higher than that of the eluent used in the subsequent step is used.
[0053]
When the motor 11 is stopped, the eluent vent 1030 is pierced by the punch 13 and the motor 11 is rotated, the eluent passes through the eluent holding container 1069, the eluent outflow channel 1070, and the nucleic acid binding site 301, It flows into the detection container 1068 (FIG. 18). The nucleic acid adsorbed on the nucleic acid binding site elutes into the eluent. As the eluent, water or a solution having a low salt concentration is used.
[0054]
When the motor 11 is stopped and the detection liquid vent 1031 is pierced by the piercing machine 13 and the motor 11 is rotated, the detection liquid flows into the detection container 1068. The detection solution is a reagent for amplifying and detecting a nucleic acid, and contains a primer, a nucleic acid synthase, deoxyribotriphosphate, a buffer, a metal ion, and the like necessary for amplification. Depending on the method of amplification or detection, it may contain a fluorescent dye, a primer modified with a fluorescent dye or biotin, or deoxyribotriphosphate modified with a fluorescent dye.
[0055]
The motor 11 is stopped, and the light emitting device 14 heats the detection container 1068 to a temperature suitable for the amplification method. For example, in the case of the constant temperature amplification method and the LAMP method, 55-65 ° C. is desirable.
After the amplification is completed, the light emitting device 14 is moved to the detection container observation window 1044, irradiated with light for detection, and the amount of emitted light or the amount of absorbed light is measured by the detection container light receiver 2014. FIG. 22 shows the relationship between the monitoring process in the middle of the entire process and the method of responding to the monitoring result.
[0056]
At the time of perforation, heating, and detection, it is necessary to stop the holding disk 12 at a predetermined position. As shown in FIG. 1, the holding disk 12 is provided with a positioning projection 17, the position detector 16 detects the rotation position of the holding disk, and rotates the motor 11, the punch 13, and the light emitting device 14. Control the irradiation.
[0057]
For example, FIG. 20 shows the operation timing of the drilling machine 13. The holding disk 12 lowers the rotation speed after the end of the flow of whole blood or each reagent, and maintains the low-speed rotation for positioning. When the position detector 16 detects the positioning projection 17, the holding disk 12 is stopped, and the punch 13 is lowered to make a hole in the lid of the vent hole of each reagent storage container, and then rise again. The post-perforation holding disk 12 rotates at such a low speed that the reagent does not flow out of the reagent storage container after the perforation, and rotates at the next analysis disk position, that is, 60 degrees when six analysis disks are mounted. Stop and repeat the same perforation operation. Where the analysis disk is mounted may be determined by, for example, irradiating light from a light emitter and examining the reflected light. After all the analysis disks have been pierced, the holding disk rotates at high speed to allow the reagent to flow.
[0058]
According to the present invention, since the process for purifying and detecting the nucleic acid of the virus contained in the sample is constantly monitored, the end of each process can be immediately sensed. , The processing time of all the steps can be reduced.
[0059]
Alternatively, according to the present invention, a high-sensitivity detector is not required for monitoring in order to quantitatively add an easily detectable detection agent instead of detecting a component contained in a sample in advance. In addition, since a fixed amount of the detection agent is added, the variation is smaller than in the case of detecting a component in a sample.
[0060]
Alternatively, according to the present invention, since an optical device for receiving light is pre-attached to the holding disk, the structure becomes optically simple, and the structure is stable, so that more accurate measurement can be performed. Can be.
[0061]
【The invention's effect】
According to the present invention, it is possible to provide a chemical analyzer for extracting and analyzing a chemical substance to be inspected from a sample in a short time.
[Brief description of the drawings]
FIG. 1 is an overall configuration diagram of a chemical analyzer according to the present embodiment.
FIG. 2 is a sectional view of a set of analysis disks according to the present embodiment.
FIG. 3 is a configuration diagram of an upper cover of an analysis disk in the present embodiment.
FIG. 4 is a configuration diagram of a channel section of an analysis disk in the present embodiment.
FIG. 5 is a configuration diagram of a lower cover of the analysis disk in the present embodiment.
FIG. 6 is an operation explanatory diagram in the present embodiment.
FIG. 7 is an operation explanatory diagram in the present embodiment.
FIG. 8 is an operation explanatory diagram in the present embodiment.
FIG. 9 is a diagram illustrating a flow state of a flow path unit in the present embodiment.
FIG. 10 is a diagram showing a flow state of a flow path unit in the present embodiment.
FIG. 11 is a diagram showing a flow state of a flow path unit in the present embodiment.
FIG. 12 is a diagram showing a flow state of a flow path unit in the present embodiment.
FIG. 13 is a sectional view of an analysis disk according to the present embodiment.
FIG. 14 is a diagram showing a flow state of a flow path section in the present embodiment.
FIG. 15 is a diagram illustrating a flow state of a flow path unit in the present embodiment.
FIG. 16 is a diagram illustrating a flow state of a flow path unit in the present embodiment.
FIG. 17 is a diagram illustrating a flow state of a flow path unit in the present embodiment.
FIG. 18 is a diagram illustrating a flow state of a flow path unit in the present embodiment.
FIG. 19 is an explanatory diagram of an operation of monitoring in the present embodiment.
FIG. 20 is an explanatory diagram of the operation of the drilling machine in the present embodiment.
FIG. 21 is a configuration diagram of a holding disk in the present embodiment.
FIG. 22 is an explanatory diagram of a monitoring operation in the present embodiment.
[Explanation of symbols]
DESCRIPTION OF SYMBOLS 1 ... Gene analysis apparatus, 2 ... Analysis disk, 11 ... Motor, 12 ... Holding disk, 13 ... Drilling machine, 14 ... Optical device, 16 ... Position detector, Reference Signs List 20 upper cover, 30 flow path, 121 projection, 1020 sample inlet, 1010 reagent inlet, 1021 vent, 301 nucleic acid binding Member, 310: sample container, 1054: serum capillary tube, 1056: mixing part, 1057: reaction container, 1068: detection container, 460: positioning hole

Claims (8)

A sample supply unit to which a sample and a label containing a specific chemical substance are supplied,
A reagent holding unit for holding a reagent,
A mixing unit in which the sample and the label are supplied from the sample supply unit, and the reagent is supplied from the reagent holding unit;
A container for a structure including a capturing unit into which the mixed fluid is introduced and capturing the chemical substance; and a detection device that is installed outside the structure and detects the sign located in the structure. And a chemical analyzer comprising: In claim 1,
The housing unit includes a mechanism for attaching and detaching the structure,
A driving device for rotationally driving the structure,
The analysis device according to claim 1, wherein the drive rotation speed is changed based on a detection value of the marker in the detection device. A structure including a sample supply unit to which a blood sample containing blood cells and serum is supplied, and a serum separation unit downstream of the sample supply unit, in which the blood cells and the serum are separated from the blood sample, can be rotated. A housing unit for housing, an attachment / detachment mechanism for the structure and the housing unit, and a detection device that is installed outside the structure and detects the sign that has flowed down into the structure,
A driving device that imparts rotation to the structure to separate the serum and the blood cells in the blood sample,
The sample supply unit is supplied with a marker flowing down in the structure together with the sample,
When the reaction of the marker detected by the detection device from the structure to which the rotation has been given is equal to or less than a predetermined value, control is performed so as to perform rotation for separating the serum and the blood cells again. Chemical analyzer. 4. The chemical analyzer according to claim 3, wherein, at the time of the second rotation, the structure is controlled so as to rotate the structure at a rotation speed higher than the rotation speed given before the detection. A sample supply unit to which a blood sample and a label including blood cells and serum are supplied,
A reagent container in which a reagent is stored, a reagent channel through which the reagent flows,
A serum separation unit formed downstream of the sample supply unit and having an area where the blood cells separated from the blood sample mainly stay and the separated serum and the label mainly stays,
A storage unit for storing a structure including the separated serum and the capillary into which the label is introduced, and a mixing unit that communicates with the reagent flow path, a detachment mechanism for attaching and detaching the structure and the storage unit, A driving device that rotates the structure, and a detection device that is installed outside the structure and detects the sign that has flowed down into the structure,
When the reaction of the marker detected by the detection device is equal to or less than a predetermined value, a control is performed such that a fluid that promotes the flow of the fluid to the capillary is supplied from an upstream side of the serum separation unit. Chemical analyzer. A sample supply unit to which a blood sample and a label including blood cells and serum are supplied,
A reagent container in which a reagent is stored, a reagent channel through which the reagent flows,
A serum separation unit formed downstream of the sample supply unit and having a region where the separated blood cells mainly stay and the separated serum and the region where the label mainly stays, including the separated serum and the label A mixing section in which the thin tube through which the fluid flows and the reagent flow path are communicated, a mixed fluid through which the mixed fluid is introduced, a nucleic acid capturing section that captures a specific nucleic acid in the mixed fluid, and the nucleic acid capturing section that passes through the nucleic acid capturing section. A liquid storage container in which a mixed fluid is stored, a storage unit that stores a structure including the structure, an attachment / detachment mechanism for the structure and the storage unit, and a driving device that rotates the structure,
A detection device that is installed outside the structure and detects the sign that has flowed down into the structure,
When the reaction of the marker detected by the detection device from the structure after performing the rotation of introducing the mixed fluid of the mixing unit into the liquid recovery container is equal to or less than a predetermined value, the mixing of the mixing unit is performed again. A chemical analyzer, which is controlled to perform rotation for introducing a fluid into the liquid recovery container. 7. The chemical analysis apparatus according to claim 6, wherein at the time of the re-rotation, the structure is controlled so that the structure is re-rotated at a rotation speed higher than the rotation speed of the rotation before the detection. 2. The chemical analyzer according to claim 1, wherein the label is at least one of a luminous body, fine particles smaller than 100 nanometers, a dye, a magnetic substance, an odor molecule, and an organic solvent.

Images