Quality measurement device and specimen treatment system

Abstract

A quality measurement device 10 measures the quality of a nucleic acid contained in a nucleic acid solution obtained during a pretreatment or after the completion of the pretreatment. The quality measurement device 10 is provided with: a PCR unit 100 for performing a PCR of a nucleic acid; a chemiluminescent unit 100A for generating chemiluminescence for the nucleic acid; a light absorption unit 100B for obtaining absorbance of the nucleic acid solution; and an electrophoresis unit 200 for performing electrophoresis of the nucleic acid. The quality measurement device 10 is provided with: a measurement body 300 for measuring fluorescence based on a nucleic acid amplified in the PCR unit 100, chemiluminescence from the chemiluminescence unit 100A, and absorbance based on transmitted light from the light absorption unit 100B; and a control unit for controlling each of the units and the measurement body.

Description

Quality measurement device and specimen processing system

[0001] The present invention relates to a quality measurement device that measures the quality of a target substance such as nucleic acid during or after pretreatment to obtain the target substance such as nucleic acid from a sample, and a sample processing system equipped with this quality measurement device.

[0002] The polymerase chain reaction (PCR), which has become widespread with the development of genetic engineering, is used to detect pathogens such as viruses. A sample containing the pathogen is collected using a commercially available collection kit, nucleic acid (DNA or RNA) is extracted from the sample, the nucleic acid is amplified by PCR, and the pathogen is tested based on the amplified nucleic acid.

[0003] The PCR method is primarily performed manually, but manual labor not only makes it difficult to continuously perform large-volume testing, but also increases the risk of operational errors and contamination during the processing. Therefore, automated PCR testing devices that automatically perform the PCR method have been proposed. For example, Precision System Science Co., Ltd. offers the geneLEAD series (Non-Patent Document 1), a fully automated PCR testing system. This fully automated PCR testing system can manually remove samples from sample containers, pre-treat them, extract and purify nucleic acids, and then perform PCR testing.

[0004] During PCR testing, a PCR cycle is repeatedly performed several tens of times using a reaction solution containing purified nucleic acids and reagents. One PCR cycle includes heating the reaction solution to a predetermined temperature and maintaining it for a predetermined period of time, and cooling the reaction solution to a predetermined temperature and maintaining it for a predetermined period of time. Repeating the PCR cycle several tens of times requires a relatively long time to complete amplification of the nucleic acids.

[0005] Therefore, in order to shorten the PCR cycle time, it is conceivable to reduce the volume of the reaction solution to shorten the time required to cool and heat the reaction solution to a predetermined temperature.Normal PCR uses a relatively large amount of reaction solution (e.g., 20 μl to 30 μl), but it is conceivable to reduce the volume of the reaction solution and perform PCR using a relatively small amount of reaction solution (e.g., 2 μl to 10 μl).

[0006] In Patent Document 1, proposed by the present inventor, a PCR device is proposed in which a PCR vessel is sealed with a lid to perform PCR. By fitting the lower part of the lid into the lower part of the vessel and sealing it, the volume of the reaction chamber is reduced and PCR is performed with a relatively small amount of reaction solution.

[0007] geneLEAD Series, [online], June 11, 2024,<URL: https: / / www.pss.co.jp / english / product / genelead / >

[0008] International Publication No. 2023 / 032870

[0009] Before performing a process such as PCR using a relatively large amount of reaction solution, it is necessary to check the quality of a target substance such as nucleic acid during or after the pretreatment. If the quality is inappropriate, the nucleic acid cannot be amplified by PCR, which may waste time during the PCR reaction. Therefore, when checking the quality of a target substance such as nucleic acid during or after the pretreatment, it is preferable to check the quality using a measurement device other than PCR that is faster or simpler. The present invention provides a quality measurement device that measures the quality of a target substance such as nucleic acid during or after the pretreatment using a relatively small amount of target substance solution such as nucleic acid.

[0010] Each example aspect of the present invention is configured as follows: (Example Aspect 1) A quality measurement device for measuring the quality of a target substance (e.g., a nucleic acid) contained in a target substance solution obtained during or after pretreatment, the quality measurement device comprising at least one or any combination of at least one PCR unit that obtains fluorescence from the target substance, at least one chemiluminescence unit that produces chemiluminescence related to the target substance, at least one absorption unit that obtains absorbance of the target substance, and at least one electrophoresis unit that performs electrophoresis of the target substance, and the quality measurement system comprises a measurement main body that measures the fluorescence from the fluorescence unit, the absorbance based on transmitted light from the absorption unit, and / or the chemiluminescence from the chemiluminescence unit, and a control unit that controls the fluorescence unit, the electrophoresis unit, and / or the measurement main body.

[0011] (Example Aspect 2) The quality measuring device according to Example Aspect 1, wherein the measurement main body comprises at least one or any combination of a fluorescence measurement unit that measures fluorescence based on the nucleic acid amplified by the fluorescence unit, an absorbance measurement unit that measures absorbance based on transmitted light from the absorbance unit, and a chemiluminescence measurement unit that measures chemiluminescence from the chemiluminescence unit. (Example Aspect 3) The quality measuring device according to Example Aspect 1, wherein the quality measuring device comprises the fluorescence unit, the absorbance unit, and the chemiluminescence unit. (Example Aspect 4) The quality measuring device according to Example Aspect 3, wherein the measurement main body comprises a PCR measurement unit that measures fluorescence based on the nucleic acid amplified by the fluorescence unit, an absorbance measurement unit that measures absorbance based on transmitted light from the absorbance unit, and a chemiluminescence measurement unit that measures chemiluminescence from the chemiluminescence unit.

[0012] (Example Aspect 5) The quality measuring device according to Example Aspect 4, wherein the measurement main body includes a scanner that moves the fluorescence measurement unit, the absorbance measurement unit, and the chemiluminescence measurement unit. (Example Aspect 6) The quality measuring device according to Example Aspect 5, wherein the scanner is connected to at least one light-projecting optical fiber for guiding excitation light from the fluorescence measurement unit to the fluorescence unit and / or for guiding incident light from the absorbance measurement unit to the absorption unit, a light-receiving optical fiber for guiding fluorescence from the fluorescence unit or chemiluminescence from the chemiluminescence unit to the fluorescence measurement unit, and a transmitted light optical fiber for guiding transmitted light from the absorption unit to the absorbance measurement unit. (Example Aspect 7) The quality measuring device according to Example Aspect 6, wherein the fluorescence measurement unit and / or the absorbance measurement unit include a plurality of light sources of different wavelengths and a plurality of light-receiving elements corresponding to the plurality of light sources, and the chemiluminescence measurement unit includes at least one light-receiving element that receives the chemiluminescence.

[0013] (Example 8) The quality measuring device according to Example 6, comprising at least one light projecting and receiving nozzle having ends of the light projecting optical fiber and the light receiving optical fiber, the light projecting and receiving nozzle being connected to the fluorescent unit, the chemiluminescent unit, and the light absorption unit via the light projecting optical fiber and the light receiving optical fiber. (Example 9) The quality measuring device according to Example 8, wherein the fluorescent unit, the chemiluminescent unit, and the light absorption unit are each composed of a container and a lid that seals the container.

[0014] (Example 10) The quality measuring device according to Example 9, wherein the container is made up of an upper container part and a lower container part that protrudes downward from the upper container part, the lower container part being formed with a smaller diameter than the upper container part, the lid is made up of an upper lid part and a lower lid part that protrudes downward from the upper lid part, the lower lid part being formed with a smaller diameter than the upper lid part, and the lower lid part is accommodated in the lower container part when the lid is attached to the container. (Example 11) The quality measuring device according to Example 10, wherein when the lower lid part is accommodated in the lower container part, the lower lid part and the lower container part come into close contact with each other to form a space that seals the nucleic acid solution.

[0015] (Example Aspect 12) The quality measuring device according to Example Aspect 9, wherein the inner surface of the container for the fluorescent unit is white, or the container for the fluorescent unit is made of a white material. (Example Aspect 13) The quality measuring device according to Example Aspect 9, wherein the inner surface of the container for the chemiluminescent unit is black, or the container for the chemiluminescent unit is made of a black material. (Example Aspect 14) The quality measuring device according to Example Aspect 9, wherein the lid includes a lid-side light-transmitting portion that transmits the excitation light, the fluorescence, and / or the chemiluminescence, and the lid-side light-transmitting portion is positioned to face the light-emitting and receiving nozzle.

[0016] (Example Aspect 15) The quality measurement device according to Example Aspect 9, wherein the container of the light absorption unit includes a container-side light transmission unit, and the container-side light transmission unit is arranged to face the end of the transmitted light optical fiber. (Example Aspect 16) The quality measurement device according to Example Aspect 1, wherein the target substance is a nucleic acid or a protein. (Example Aspect 17) The quality measurement device according to Example Aspect 16, wherein the fluorescence unit is a PCR unit that performs a PCR reaction of the nucleic acid. (Example Aspect 18) The quality measurement device according to Example Aspect 1, wherein the control unit performs qualitative and / or quantitative processing of the target substance using the measured fluorescence, chemiluminescence, and / or absorbance.

[0017] (Example Aspect 19) The quality measurement device according to Example Aspect 18, further comprising an imaging unit that images an electrophoresis pattern obtained from the electrophoresis, wherein the control unit executes a process for evaluating the quality of the target substance based on the electrophoresis pattern. (Example Aspect 20) A specimen processing system, comprising: a specimen processing device that performs a process for extracting and purifying nucleic acid from a specimen and a process for amplifying the purified nucleic acid; and the quality measurement device according to Example Aspect 1. (Example Aspect 21) The specimen processing system according to Example Aspect 20, further comprising a transfer device that transfers the nucleic acid solution from the specimen processing device to the quality measurement device.

[0018] (Example 22) A sequencer that sequences nucleic acids obtained from a sample, comprising: a sample processing device having a pre-treatment section that extracts and purifies the nucleic acids from the sample, a pre-treatment side PCR section that amplifies the nucleic acids by PCR using a nucleic acid solution obtained from the pre-treatment section, and a first nozzle unit that moves the nucleic acid solution; a quality measurement device according to Example 1 that measures the quality of the nucleic acids using the nucleic acid solution obtained from the pre-treatment section; a transfer device that transfers the nucleic acid solution from the pre-treatment section to the quality measurement device; a flow cell section that sequences the nucleic acids using the nucleic acid solution; and a control section that controls processing in the sample processing device, the quality measurement device, the transfer device, and the flow cell section, wherein the sequencer sequences the nucleic acids when the control section determines that the quality of the nucleic acids measured by the quality measurement device satisfies at least one predetermined condition. (Example 23) A sequencer according to Example 22, wherein the pre-processing unit comprises a nucleic acid extraction unit that extracts nucleic acid from the sample, a sizing unit that sizes the extracted nucleic acid, and an adapter binding unit that binds adapters to both ends of the sized nucleic acid.

[0019] The quality measurement device of the present invention can inspect the quality of target substances such as nucleic acids using a relatively small amount of target substance solution such as nucleic acids during or after preprocessing, before performing a process such as PCR using a relatively large amount of target substance solution such as nucleic acids.

[0020] 17 is a top perspective view showing a sample processing system according to an embodiment of the present invention. FIG. 18 is a top view showing the sample processing system of FIG. 1. FIG. 19 is an top perspective view showing the lid and container of the PCR unit of FIG. 1. FIG. 20 is a longitudinal sectional view showing the PCR unit of FIG. 1. FIG. 21 is a longitudinal sectional view showing a modified PCR unit of FIG. 1. FIG. 22 is an top perspective view showing the lid and container of the chemiluminescence unit of FIG. 1. FIG. 21 is a longitudinal sectional view showing a modified chemiluminescence unit of FIG. 1. FIG. 22 is an top perspective view showing the lid and container of the light absorption unit of FIG. 1. FIG. 23 is a longitudinal sectional view showing the light absorption unit of FIG. 1. FIG. 24 is a longitudinal sectional view showing a modified light absorption unit of FIG. 1. FIG. 25 is a plan view and FIG. 26 is a cross-sectional view showing a stage of the quality measurement device of FIG. 1. FIG. 26 is a perspective view showing the measurement main body of FIG. 1, with the scanning body of the measurement main body located on the rear left side. FIG. 27 is a perspective view showing the measurement main body of FIG. 1, with the scanning body of the measurement main body located on the front right side. FIG. 28 is a top view of the measurement main body of FIG. 11. FIG. 29 is a side view showing the sample processing system of FIG. 1. FIG. 29 is a bottom perspective view showing the second nozzle unit of FIG. 1. FIG. 29 is a bottom perspective view showing the light emitting and receiving nozzle of FIG. 17.

[0021] An embodiment of the quality inspection device and a sample processing system equipped with the same of the present invention will be described with reference to the drawings. Note that in each figure, identical parts are designated by the same reference numerals, and their description will be omitted as appropriate. In the present invention, a relatively small amount of target substance solution, such as nucleic acid, may be smaller than a relatively large amount of target substance solution, such as nucleic acid. In this embodiment, although not limited thereto, the relatively large amount of target substance solution, such as nucleic acid, may be, for example, 20 to 30 μl, and the relatively small amount of target substance solution, such as nucleic acid, may be, for example, 2 to 10 μl. Note that, in this embodiment, the target substance whose quality is inspected is nucleic acid, but this is not limited thereto, and any biologically related substance (e.g., protein) may be used.

[0022] 1 and 2, the sample processing system 1 (sequencer) of this embodiment is composed of a sample processing and measurement device 2, a quality inspection device 10, and a pitch-converting movement device 400. The sample processing and measurement device 2 is composed of a pretreatment unit (pretreatment device) 20 that performs pretreatment on samples such as blood or body fluids to obtain a nucleic acid solution, a pretreatment-side PCR unit 25 that performs a PCR reaction on a relatively large amount of the nucleic acid solution obtained after pretreatment, a flow cell unit 21 that performs sequencing, and a first nozzle unit 30 that moves in the x direction above the pretreatment unit 20, flow cell unit 21, and pretreatment-side PCR unit 25. The first nozzle unit 30 has a plurality of dispensing nozzles aligned in the y direction.

[0023] As shown in Fig. 1, the pitch-changing transfer device 400 includes a plurality of nucleic acid solution containers 401 that contain relatively small amounts of nucleic acid solution obtained during or after pretreatment. The pitch-changing transfer device 400 transfers the plurality of nucleic acid solution containers 401 from the pretreatment unit 20 to the quality measurement device 10. The pitch-changing transfer device 400 moves along rails extending in the y direction, and can be in a large pitch state in which it is located on the sample processing and measurement device 2 side, and a small pitch state in which it is located on the quality inspection device 10 side, as shown in Fig. 1. The large pitch state only needs to have a larger pitch than the small pitch state, and the size of each pitch is not limited.

[0024] The quality measurement device 10 measures the quality of a relatively small amount of nucleic acid solution obtained during or after pretreatment. As shown in Figure 1, the quality measurement device 10 is composed of a PCR section (quality measurement side PCR section) 100, a chemiluminescence section 100A, an absorption section 100B, an electrophoresis section 200, a second nozzle unit 60 that moves in the x direction above each section 100, 100A, 100B, and a measurement body 300 that measures light received by each section 100, 100A, 100B. Note that optical fibers, which will be described later, are not shown in Figure 1.

[0025] The quality measurement device 10 includes, on a stage 11, one or any combination of at least one PCR unit 100 (FIG. 4 or 5) that amplifies a nucleic acid solution by a real-time PCR reaction, at least one chemiluminescence unit 100A (FIG. 7 or 8) that chemiluminesces an immunoreaction solution of nucleic acid, at least one light-absorbing unit 100B (FIG. 10 or 11) that obtains transmitted light through the nucleic acid solution to measure the absorbance of the nucleic acid solution, and at least one electrophoresis unit 200 (FIG. 1) that performs electrophoresis of the nucleic acid solution. The quality measurement device 10 can preferably include, on the stage 11, the PCR unit 100, the chemiluminescence unit 100A, the light-absorbing unit 100B, and the electrophoresis measurement unit 200.

[0026] The PCR section (fluorescence section) 100 according to this embodiment will be described with reference to Figures 3 to 5. The PCR section 100 comprises a PCR vessel (measurement vessel) 110 having at least a white inner surface, a PCR vessel lid (common cap) 120, a thermal cycler 130, and a light-emitting / receiving nozzle 40 or 40A.

[0027] The PCR vessel 110 is preferably made of white resin. The PCR vessel 110 is composed of an upper vessel portion 111 that houses a middle lid portion 123 of the lid 120, and a lower vessel portion 112 that houses a lower lid portion 121 of the lid 120 and a reagent and nucleic acid solution. The upper vessel portion 111 is preferably cylindrical. The lower vessel portion 112 is formed with a smaller diameter than the upper vessel portion 111. The lower vessel portion 112 preferably protrudes downward in a conical shape relative to the upper vessel portion 111. The lower vessel portion 112 preferably has a rounded lowest vessel portion 112a.

[0028] The lid 120 is composed of a cylindrical upper lid part 121 that houses the lower end side of the light emitting / receiving nozzle 40, a cylindrical middle lid part 123 that is housed in the upper container part 111, and a lower lid part 122 that is connected to the annular inclined surface of the middle lid part 123. The lower lid part 122 protrudes downward in a conical shape relative to the upper lid part 121. The lower lid part 22 is formed with a smaller diameter than the upper lid part 121 and the middle lid part 123.

[0029] A flat light-transmitting portion (lid-side light-transmitting portion) 122a is formed at the lower end of the lid lower portion 122. The lid lower portion 122 is preferably tapered. At least the light-transmitting portion 122a of the lid lower portion 122 is formed from a transparent material. The transparent material can be a resin that transmits excitation light, fluorescence, and / or chemiluminescence, as described below. The outer surface of the conical lid lower portion 122 is in close contact with the inner surface of the conical container lower portion 112, forming a space S (FIG. 4) for containing the reagent and nucleic acid solution between the light-transmitting portion 122a and the container lowest portion 112a.

[0030] 4 , with the lid lower portion 122 accommodated in the container lower portion 112, the lid lower portion 122 and the container lower portion 113 come into close contact with each other, sealing the nucleic acid solution in the space S. In other words, when the lid lower portion 122 is accommodated in the container lower portion 112 with the lid 120 attached to the container 110, the nucleic acid solution dispensed into the container lower portion 112 overflows from between the lid lower portion 122 and the container lower portion 112, expelling the air between the lid lower portion 122 and the container lower portion 112 upward. This allows a relatively small amount of nucleic acid solution to be sealed in the space S without an air layer. Because the space S does not contain an air layer, condensation does not form on the surface of the light transmitting portion 122a during PCR.

[0031] The light-emitting and receiving nozzle 40 is provided in a second nozzle unit 60 ( FIG. 1 ), which is disposed above the quality measuring device 10 so as to be movable in the x direction by a second nozzle unit movement mechanism (e.g., a motor and a rail). A detection end movement mechanism (e.g., a motor and an actuator) moves the light-emitting and receiving nozzle 40 in the z direction. A lid 120 can be attached to the light-emitting and receiving nozzle 40. With the lid 120 attached to the light-emitting and receiving nozzle 40, the light-emitting and receiving nozzle 40 can be moved by the second nozzle unit movement mechanism and the detection end movement mechanism.

[0032] 4 is composed of a light-projecting optical fiber 41a, a light-receiving optical fiber 41b, an optical lens 42 disposed at the lower ends of the optical fibers 41a and 41b, and an end cover 43 that houses the optical fibers 41a and 41b and the optical lens 42. The lower surface of the optical lens 42 is exposed inside the lid 120. The space between the lid 120 and the light-projecting and receiving nozzle 40 is preferably sealed with a sealant or the like to prevent the optical lens 42 from fogging up.

[0033] The direct light emitting and receiving type light emitting and receiving nozzle 40A shown in Figure 5 is interchangeable with the light emitting and receiving nozzle 40 shown in Figure 4. Regarding the light emitting and receiving nozzle 40A, only the parts that differ from the light emitting and receiving nozzle 40 will be described. The light emitting and receiving nozzle 40A does not have an optical lens 42, and the lower end 41a1 of the light emitting optical fiber 41a and the lower end 41b1 of the light receiving optical fiber 41b are arranged so as to face the light transmitting portion 122a near the light transmitting portion 122a. The lower end 41a1 and the lower end 41b1 are preferably arranged so as to abut against the light transmitting portion 122a.

[0034] The thermal cycler 130 performs a PCR reaction by repeatedly heating and cooling a nucleic acid solution contained in the space S, which contains a nucleic acid amplification reagent and a measurement reagent such as a fluorescent dye, to amplify the nucleic acid. The amplified nucleic acid binds to the fluorescent dye and is excited by excitation light projected from the light-projecting optical fiber 41a, emitting fluorescence. This fluorescence is received by the light-receiving optical fiber 41b. The measurement main body 300, which will be described later, supplies excitation light to the light-projecting optical fiber 41a and measures the fluorescence from the light-receiving optical fiber 41b. The PCR unit 100 preferably performs real-time PCR.

[0035] [Chemiluminescence Unit] A chemiluminescence unit 100A for quality confirmation according to this embodiment will be described using Figures 6 to 8. Regarding the chemiluminescence unit 100A, differences from the PCR unit 100 will be described, and the same parts will be assigned the same reference numerals and will not be described again. The chemiluminescence unit 100A does not have a thermal cycler 130, but has a light-blocking container 110A. The container 110A is preferably formed from a light-blocking material (e.g., black resin), or its inner surface is coated with a light-blocking paint (black paint).

[0036] The light-emitting / receiving nozzles 40, 40A have an annular portion 44 (annular protrusion) formed or arranged in an annular shape on the outer periphery of the end cover 43, and the annular portion 44 has an annular light-shielding groove 41a that engages with the annular upper end of the lid upper portion 121. The lid 120, except for the light-transmitting portion 122a, is preferably formed from a light-shielding material (preferably black resin) or can have a light-shielding coating layer (black coating layer) on its inner surface.

[0037] The space S of the chemiluminescence unit 100A contains a nucleic acid solution and a chemiluminescent substance that emits light through an antigen-antibody reaction with nucleic acid (a predetermined base sequence). The light projecting / receiving nozzles 40, 40A do not supply excitation light from the light projecting optical fiber 41a, but transmit the chemiluminescence received by the light receiving optical fiber 41b to the measurement body 300. A photodetector provided in the measurement body 300 measures the chemiluminescence transmitted from the light receiving optical fiber 41b. The photodetector may be, for example, a photomultiplier tube (PMT).

[0038] [Light-Absorption Unit] The light-absorption unit 100B according to this embodiment will be described using Figures 9 to 11. Regarding the light-absorption unit 100B, only the parts that differ from the PCR unit 100 will be described, and the same parts will be assigned the same reference numerals and will not be described again. The light-absorption unit 100B does not have a thermal cycler 130, but has a container 110B equipped with a light-transmitting unit 113B (container-side light-transmitting unit), and a lower measurement end 50.

[0039] The light-transmitting portion 113B is formed at the lower end of the container 110B. The light-transmitting portion 113B is made of a transparent material and has a flat transmitting surface. The transparent material can preferably be a transparent resin that transmits visible light (incident light). The lower detection end portion 50 is composed of an end portion of the transmitted light optical fiber 51 and a cover 53 that covers the circumferential surface of the transmitted light optical fiber 51. The upper end surface 51a of the transmitted light optical fiber 51 faces the light-transmitting portion 113B.

[0040] A nucleic acid solution is contained in space S of light absorption unit 100B. Light projecting and receiving nozzles 40, 40A supply incident light (e.g., visible light) from light projecting optical fiber 41a to the nucleic acid solution in space S, and transmitted light that has passed through the nucleic acid solution and 113B is received by upper end surface 51a of transmitted light optical fiber 51. The transmitted light is sent to measurement body 300 through transmitted light optical fiber 51, and a light receiving element of measurement body 300 measures the absorbance or turbidity of the transmitted light.

[0041] [Quality Measurement Device] As shown in Figure 12, the quality measurement device 10 has multiple processing lines (e.g., four processing lines) extending in the x direction on a flat quality measurement stage 11. Each processing line has multiple dispensing tips 12, at least one punching tip 13, components of a PCR section 100, components of a chemiluminescence section 100A, components of an absorption section 100B, and an electrophoresis measurement section 200. A second nozzle unit 60 (Figures 1 and 2) moves in the x direction above the quality measurement device 10. To the side (right side) of the quality measurement stage 11, multiple nucleic acid solution containers 401 that move in the y direction by a pitch conversion mechanism 400 are arranged corresponding to each processing line.

[0042] The components of the PCR unit 100 are a PCR reagent container 130, a lid 120, and a PCR container 110, which are arranged in a line in the x direction. As shown in FIG. 12(b), a thermal cycler 130 is arranged below the PCR container 110. A dispensing nozzle 65 (FIG. 16) of the second nozzle unit 60 transfers PCR reagent from a PCR reagent container 130A to the PCR container 110. The dispensing nozzle 65 transfers a relatively small amount of nucleic acid solution from a nucleic acid solution container 401 of the pitch conversion mechanism 400 to the PCR container 110. The light-emitting / receiving nozzle 40 or 40A fits the lid 120 onto the PCR container 110.

[0043] The components of the chemiluminescence unit 100A are a chemiluminescent reagent container 130A, a lid 120, and a light-blocking (black) chemiluminescent container 110A, which are arranged in a row in the x direction. A dispensing nozzle 65 transfers the chemiluminescent reagent from the chemiluminescent reagent container 130A to the chemiluminescent container 110A. The dispensing nozzle 65 transfers a relatively small amount of nucleic acid solution from a nucleic acid solution container 401 of the pitch conversion mechanism 400 to the chemiluminescent container 110A. The light emitting / receiving nozzle 40 or 40A fits the lid 120 into the chemiluminescent container 110A.

[0044] The components of the light-absorbing unit 100B are a container 130B for absorbance measurement reagent, a lid 120, and a light-absorbing container 110B, which are arranged in a row in the x direction. As shown in FIG. 12(b), a lower measurement end portion 50 (optical fiber 51 for transmitted light) is arranged below the light-absorbing container 110B. A dispensing nozzle 65 transfers the absorbance measurement reagent from the container 130B for absorbance measurement reagent to the light-absorbing container 110B. The dispensing nozzle 65 transfers a relatively small amount of nucleic acid solution from a nucleic acid solution container 401 of the pitch conversion mechanism 400 to the light-absorbing container 110B. The light-emitting / receiving nozzle 40 or 40A fits the lid 120 into the light-absorbing container 110B.

[0045] The components of the electrophoresis measurement unit 200 include an electrophoresis reagent container 230 and an electrophoresis unit 200. The electrophoresis unit 200 has one or more solution dispensing ports 201 corresponding to one or more processing lanes, an electrophoresis gel 203, and a pair of electrophoresis electrodes 205 that sandwich the electrophoresis gel 203. A dispensing nozzle 65 transfers the electrophoresis reagent from the electrophoresis reagent container 230 to the electrophoresis unit 200. The dispensing nozzle 65 transfers a relatively small amount of nucleic acid solution from a nucleic acid solution container 401 of the pitch conversion mechanism 400 to the solution dispensing port 201. An electrophoresis control unit (not shown) applies a voltage between the pair of electrophoresis electrodes 205 to perform electrophoresis.

[0046] 13 to 15, the measurement main body 300 is described. The measurement main body 300 is composed of a PCR measurement section (fluorescence measurement section) 310 that measures the fluorescence during real-time PCR obtained from the PCR section 100, an absorbance measurement section 320 that measures the transmitted light obtained from the light absorption section 100B, a chemiluminescence measurement section 330 that measures the chemiluminescence obtained from the chemiluminescence section 100A, a scanning body 340 (optical fiber end holder) that can move in the direction of the arrow while facing each section, a scanning body moving mechanism 350 that moves the scanning body 340, and a control section (CPU) 360 that controls each measurement section and the scanning mechanism.

[0047] The scanning body 340 moves along a guide rail 370 to a position facing each of the PCR measurement unit 310, the absorbance measurement unit 320, and the chemiluminescence measurement unit 330. The scanning body 340 is connected to a first optical fiber group A, a second optical fiber group B, and a third optical fiber group C.

[0048] The first optical fiber group A, as shown in Figures 15 and 16, is a plurality of light-emitting optical fibers 41a connected to the plurality of light-emitting and light-receiving nozzles 40 or 40A of the second nozzle unit 60, and transmits fluorescence from the light source of the PCR measurement section 310 to the space S of the PCR section 100. The second optical fiber group B, as shown in Figures 15 and 16, is a plurality of light-receiving optical fibers 41b connected to the light-emitting and light-receiving nozzles 40 or 40A, and transmits fluorescence or chemiluminescence obtained from the PCR section 100 or chemiluminescence section 100A to the PCR measurement section 310 or chemiluminescence measurement section 330. The third optical fiber group C, as shown in Figures 15 and 16, is a plurality of transmitted light optical fibers 51 provided below the light-absorbing section 100B, and transmits transmitted light obtained from the light-absorbing section 100B to the absorbance measurement section 320.

[0049] 13 and 14, the scanning body moving mechanism 350 is composed of a drive motor 351, a timing belt 353, a pair of pulleys 354 that rotatably support the timing belt 353, and a connecting plate 355 that is moved by the timing belt 353. One of the pair of pulleys 354 is rotated by the drive motor 351. The scanning body 340 moves integrally with the connecting plate 355.

[0050] The measuring body 300 of this embodiment can preferably perform multiplexed and continuous measurements of the multiple PCR units 100, the multiple chemiluminescent units 100A, and the multiple absorber units 100B shown in Figure 12 by moving the scanning mechanism 350.

[0051] The measurement body 300 has a scanning line LA of the first optical fiber group A (light-projecting optical fiber 41 a), which is shown by a dashed line in Fig. 15. The absorbance measurement unit 320 has positions for arranging a plurality of light sources on the scanning line LA. The absorbance measurement unit 320 preferably has, linearly on the scanning line LA, a light source position P1 where a first light source (first wavelength region light source) is arranged, a light source position P2 where a second light source (second wavelength region light source) is arranged, and a light source position P3 where a third light source (third wavelength region light source) is arranged.

[0052] The PCR measurement unit 310 has positions for arranging multiple excitation light sources on the scanning line LA. The PCR measurement unit 310 preferably has an excitation light position P4 where a first excitation light source is arranged, an excitation light position P5 where a second excitation light source is arranged, and an excitation light position P6 where a third excitation light source is arranged, arranged linearly on the scanning line LA. The measurement body 300 has a scanning line LB of the second optical fiber group B (light-receiving optical fiber 41b), shown by a dashed line in Figure 15. The chemiluminescence measurement unit 330 has a position P7 on the scanning line LB where at least one phototransducer (PMT) is arranged.

[0053] The measurement body 300 has a scanning line LC of the third optical fiber group C (transmitted light optical fibers 51), shown by a dashed line in Fig. 15. The absorbance measurement unit 320 has positions for arranging a plurality of transmitted light receiving elements on the scanning line LC. The absorbance measurement unit 320 preferably has a position P8 for a fourth light receiving element, a position P9 for a fifth light receiving element, and a position P10 for a sixth light receiving element linearly arranged on the scanning line LC.

[0054] The PCR measurement unit 310 has positions for arranging a plurality of light receiving elements on the scanning line LB. The PCR measurement unit 310 preferably has a linear arrangement on the scanning line LB of a light receiving position P11 where a first light receiving element (light receiving element of a first wavelength) is arranged, a light receiving position P12 where a second light receiving element (light receiving element of a second wavelength) is arranged, and a light receiving position P13 where a third light receiving element (light receiving element of a third wavelength) is arranged.

[0055] The second nozzle unit 60 will be described using Figures 16 and 17. The second nozzle unit 60 includes a plurality of light-emitting and light-receiving nozzles 40 or 40A aligned in a line in the y direction, a first removal mechanism 63 that pushes down and removes a plurality of lids 120 attached to the plurality of light-emitting and light-receiving nozzles 40 or 40A, and a second removal mechanism 67 that pushes down and removes a plurality of dispensing nozzles 65 aligned in a line in the y direction and a plurality of dispensing tips 12 attached to the plurality of dispensing nozzles 65. The first removal mechanism 63 and the second removal mechanism 67 are each composed of a removal plate and a motor that drives the removal plate. As shown in Figure 18, the first removal mechanism 63 includes a light-shielding groove 44a around the periphery of the light-emitting and light-receiving nozzle 40.

[0056] [Sample Processing and Measuring Apparatus] The sample processing and measuring apparatus 2 will be described with reference to Figure 2. The sample processing and measuring apparatus 2 is composed of a pretreatment section 20, a flow cell section 21, a pretreatment side PCR section 25, and a first nozzle unit 30. The pretreatment section 20 is composed of a consumables storage section 22, a nucleic acid extraction and purification cartridge 23a (nucleic acid extraction section), a nucleic acid sizing cartridge 23b (sizing section), and a ligation cartridge 23c (ligation section). The consumables storage section 22 stores consumables such as one or more dispensing tips, one or more sample tubes, and / or one or more perforation tips.

[0057] The nucleic acid extraction and purification cartridge 23a includes a well for containing a nucleic acid extraction and purification reagent and a well for nucleic acid extraction and purification. The nucleic acid sizing cartridge 23b is composed of a well for containing a nucleic acid sizing reagent and a well for performing nucleic acid sizing, and performs sizing of the nucleic acid extracted using the nucleic acid sizing reagent. The ligation cartridge 23c is composed of a well for containing a ligation reagent and a well for performing ligation (adapter binding), and binds adapters to both ends of the sized nucleic acid. The wells for containing each reagent are pre-sealed with freeze-dried reagents using aluminum seals or the like. The PCR unit 25 is composed of a PCR cartridge for containing a PCR well and a PCR reagent, and a thermal cycler installed below the PCR well.

[0058] If the control unit determines that the quality of the nucleic acids obtained from the quality measurement device 10 after preprocessing satisfies at least one of the predetermined conditions described below, the flow cell unit 21 performs sequencing of the nucleic acids (adapter-bound nucleic acids) using a relatively large amount of nucleic acid solution. The flow cell unit 21 has clusters of nucleic acids immobilized via adapters for binding the nucleic acids. The flow cell unit performs simultaneous parallel sequencing of multiple nucleic acids immobilized on the flow cell using four types of fluorescently labeled nucleotides and a polymerase enzyme based on a single-base synthesis technique known as sequencing by synthesis (SBS). The flow cell unit is equipped with an excitation light source and a fluorescence receiving element (not shown), and the control unit controls the excitation light source and the fluorescence receiving element.

[0059] [Real-time PCR Measurement by Measurement Main Body] The operation of the PCR measurement unit 310 of the measurement main body 300 when measuring fluorescence obtained from multiple PCR units 100 will be described. The multiple PCR units 100 are lined up in the y direction as shown in FIG. 12(a). Multiple nucleic acid solutions obtained from multiple different samples are contained in spaces S within the multiple PCR units 100. The multiple light-emitting and receiving nozzles 40 or 40A of the second nozzle unit 60 are moved along the x direction above the container 110 with the lid 120 attached, to position the PCR unit 100 in a state where measurement is possible, as shown in FIG.

[0060] While PCR is being performed in the state shown in FIG. 4 , the scanner 340 scans the PCR measurement unit 310 along scanning lines LA and LB shown in FIG. 15 while projecting and receiving light into the multiple PCR units 100. The light-projecting optical fiber 41a and the light-receiving optical fiber 41b for each PCR unit 100 are paired and arranged side by side approximately perpendicular to the scanning direction (or vertically) in groups A and B shown in FIG. 15 . Therefore, when the scanner 340 scans excitation light positions P4, P5, and P6 of the PCR measurement unit 310 along scanning line LA and light-receiving positions P11, P12, and P13 of the PCR measurement unit 310 along scanning line LB, excitation light of different wavelengths can be supplied to the multiple PCR units 100, and fluorescence corresponding to the excitation light of different wavelengths can be measured. The control unit 360 performs qualitative and / or quantitative processing of a target substance such as a nucleic acid based on the fluorescence measured by the PCR measurement unit (fluorescence measurement unit) 310. The control unit 360 determines whether the quality (fluorescence intensity) of the measured target substance satisfies a predetermined condition. If the control unit 360 determines that the predetermined condition (fluorescence intensity condition) is satisfied, it registers in a storage unit (such as a hard disk) that the quality satisfies the condition.

[0061] [Absorbance Measurement by Measurement Main Body] The operation of the absorbance measurement unit 320 of the measurement main body 300 when measuring absorbance (transmitted light) obtained from multiple light-absorbing units 100B will be described. The multiple light-absorbing units 100B are lined up in the y direction as shown in FIG. 12 . Multiple nucleic acid solutions obtained from multiple different samples are respectively contained in spaces S within the multiple light-absorbing units 100B. The multiple light-emitting and receiving nozzles 40 of the second nozzle unit 60 are moved along the x direction onto the container 110 with the lid 120 attached, and the inside of the light-absorbing unit 100B is positioned so that measurement is possible, as shown in FIG. 10 or 11 .

[0062] 10 or 11 , light is projected from above into the plurality of light-absorbing units 100B, and the light that passes through the space S is received from below while the scanner 340 scans the absorbance measurement unit 320 along scanning lines LA and LC. The light-projecting optical fiber 41a and the transmitted light optical fiber 51 for each light-absorbing unit 100B are paired and arranged substantially perpendicular to the scanning direction (or vertically) in groups A and C in FIG. 15 . Therefore, when the scanner 340 scans the visible light positions P1, P2, and P3 of the absorbance measurement unit 320 along scanning line LA and the light-receiving positions P8, P9, and P10 of the PCR measurement unit 310 along scanning line LB, different types of visible light are supplied to the plurality of light-absorbing units 100B, and transmitted light corresponding to the different types of visible light is received, thereby measuring the absorbance. The control unit 360 can calculate turbidity from the measured absorbance. The control unit 360 performs qualitative and / or quantitative processing of the target substance, such as a nucleic acid, based on the absorbance measured by the absorbance measurement unit 320. The control unit 360 determines whether the quality (absorbance or turbidity) of the measured target substance satisfies predetermined conditions. If the control unit 360 determines that the predetermined conditions (absorbance or turbidity conditions) are met, it registers in the storage unit that the quality meets the conditions.

[0063] [Chemiluminescence Measurement by Measurement Main Body] The operation of the chemiluminescence measurement unit 330 of the measurement main body 300 when measuring chemiluminescence obtained from multiple chemiluminescence units 100A will be described. The multiple chemiluminescence units 100A are lined up in the y direction as shown in FIG. 12 . Multiple nucleic acid solutions obtained from multiple different samples are contained in spaces S within the multiple chemiluminescence units 100A. The multiple light-emitting and receiving nozzles 40 or 40A of the second nozzle unit 60 are moved along the x direction above the container 110 with the lid 120 attached, and the inside of the chemiluminescence unit 100A is positioned so that measurement is possible, as shown in FIG. 7 or 8 .

[0064] In this state, the scanner 340 is caused to scan over the chemiluminescence measurement unit 330 along the scanning line LB in FIG. 15 while receiving chemiluminescence from the multiple chemiluminescent units 100A. When the scanner 340 scans the light-receiving position P7 of the chemiluminescent measurement unit 330 along the scanning line LB, the chemiluminescence obtained from each of the multiple chemiluminescent units 100A can be measured. The control unit 360 performs qualitative and / or quantitative processing of the target substance, such as a nucleic acid, based on the chemiluminescence measured by the chemiluminescent measurement unit 313. The control unit 360 determines whether the quality (chemiluminescence intensity) of the measured target substance satisfies predetermined conditions. If the control unit 360 determines that the predetermined conditions (chemiluminescence intensity conditions) are met, it registers in the memory unit that the quality satisfies the conditions.

[0065] [Electrophoresis Measurement by Electrophoresis Unit] As shown in FIG. 12 , the electrophoresis unit 200 has multiple solution dispensing ports 201 arranged in the y direction. Multiple nucleic acid solutions obtained from multiple different samples are dispensed into the multiple solution dispensing ports 201 using multiple dispensing nozzles 65. After dispensing the nucleic acid solutions, a voltage is applied between a pair of electrophoresis electrodes 205, causing the nucleic acids to migrate along the electrophoresis gel 203. After a predetermined time has passed, an electrophoresis pattern generated by the migration of the nucleic acids is captured by a photographing unit (an image sensor such as a CCD). The control unit 360 can perform image recognition of the captured electrophoresis pattern and perform a quality evaluation process for the target substance, such as nucleic acid. The photographing unit can be provided in the second nozzle unit 60. The control unit 360 determines whether the quality (electrophoresis pattern) of the measured target substance satisfies predetermined conditions. If the control unit 360 determines that the predetermined conditions (electrophoresis conditions) are met, it registers that the quality satisfies the conditions.

[0066] Although the embodiments of the present invention have been described with respect to measuring the quality of nucleic acids during or after pretreatment, the quality measurement device of the present invention is not limited to measuring the quality of nucleic acids using PCR reactions. The quality measurement device of the present invention can also be applied to various target substances (e.g., proteins, etc.) obtained during or after pretreatment, such as fluorescence, chemiluminescence measurement using antigen-antibody reactions (immunoassays), absorbance measurement using various absorbance settings, and / or electrophoresis pattern measurement based on the size of charged target substances. Furthermore, the quality measurement device of the present invention can perform multiplexed, highly sensitive quantitative and / or qualitative processing of target substances using the obtained measurements.

[0067] REFERENCE SIGNS LIST 1 Sample processing system (fully automatic sequencer) 2 Sample processing and measurement device 10 Quality measurement device 20 Pretreatment unit 21 Flow cell unit 30 First nozzle unit 40 Light emitting and receiving nozzle 60 Second nozzle unit 100 PCR unit 100A Chemiluminescence unit 100B Absorption unit 200 Electrophoresis unit 300 Measurement main body 310 PCR measurement unit 320 Absorbance measurement unit 330 Chemiluminescence measurement unit 340 Scanning body 400 Pitch conversion type moving device

Claims

1. A quality measuring device for measuring the quality of a target substance (e.g., a nucleic acid) contained in a target substance solution obtained during or after completion of pretreatment, said quality measuring device comprising at least one or any combination of at least one fluorescence unit that obtains fluorescence from the target substance, at least one chemiluminescence unit that produces chemiluminescence related to the target substance, at least one absorption unit that obtains absorbance of the target substance solution, and at least one electrophoresis unit that performs electrophoresis of the target substance, and said quality measuring system comprising a measurement main body that measures the fluorescence from the fluorescence unit, the absorbance based on the transmitted light from the absorption unit, and / or the chemiluminescence from the chemiluminescence unit, and a control unit that controls the fluorescence unit, the electrophoresis unit, and / or the measurement main body.

2. A quality measuring device as described in claim 1, wherein the measurement body comprises at least one or any combination of a fluorescence measuring unit that measures the fluorescence from the fluorescent unit, an absorbance measuring unit that measures the absorbance based on the transmitted light from the light absorption unit, and a chemiluminescence measuring unit that measures the chemiluminescence from the chemiluminescent unit.

3. A quality measuring device according to claim 1, comprising the fluorescent unit, the light absorbing unit, and the chemiluminescent unit.

4. A quality measuring device as described in claim 3, wherein the measurement body comprises a PCR measurement unit that measures fluorescence based on the nucleic acid amplified in the fluorescent unit, an absorbance measurement unit that measures absorbance based on the transmitted light from the absorbance unit, and a chemiluminescence measurement unit that measures chemiluminescence from the chemiluminescence unit.

5. A quality measuring device according to claim 4, wherein the measurement body comprises a scanner which moves the fluorescence measuring section, the absorbance measuring section, and the chemiluminescence measuring section.

6. A quality measuring device as described in claim 5, wherein the scanning body is connected to at least one light-projecting optical fiber for guiding excitation light from the fluorescence measurement unit to the fluorescence unit and / or for guiding incident light from the absorbance measurement unit to the absorption unit, is connected to a light-receiving optical fiber for guiding fluorescence from the fluorescence unit or chemiluminescence from the chemiluminescence unit to the fluorescence measurement unit, and is connected to a transmitted light optical fiber for guiding transmitted light from the absorption unit to the absorbance measurement unit.

7. A quality measuring device as described in claim 6, wherein the fluorescence measuring unit and / or the absorbance measuring unit comprises a plurality of light sources having different wavelengths and a plurality of light receiving elements corresponding to the plurality of light sources, and the chemiluminescence measuring unit comprises at least one light receiving element that receives the chemiluminescence.

8. A quality measuring device as described in claim 6, comprising at least one light projecting and receiving nozzle having ends of the light projecting optical fiber and the light receiving optical fiber, the light projecting and receiving nozzle being connected to the fluorescent unit, the chemiluminescent unit, and the light absorbing unit via the light projecting optical fiber and the light receiving optical fiber.

9. A quality measuring device according to claim 8, wherein the fluorescent unit, the chemiluminescent unit, and the light absorbing unit each comprise a container and a lid for sealing the container.

10. A quality measuring device as described in claim 9, wherein the container is composed of an upper container part and a lower container part that protrudes downward from the upper container part, the lower container part being formed with a smaller diameter than the upper container part; the lid is composed of an upper lid part and a lower lid part that protrudes downward from the upper lid part, the lower lid part being formed with a smaller diameter than the upper lid part; and when the lid is attached to the container, the lower lid part is received in the lower container part.

11. A quality measuring device according to claim 10, wherein when the lower part of the lid is placed in the lower part of the container, the lower part of the lid and the lower part of the container come into close contact with each other to form a space that seals the nucleic acid solution.

12. A quality measuring device according to claim 9, wherein the inner surface of the container of the phosphor is white or the container of the phosphor is made of a white material.

13. A quality measuring device according to claim 9, wherein the inner surface of the container of the chemiluminescent unit is black, or the container of the chemiluminescent unit is formed from a black material.

14. A quality measuring device as described in claim 9, wherein the lid has a lid-side light-transmitting portion that transmits the excitation light, the fluorescence, and / or the chemiluminescence, and the lid-side light-transmitting portion is positioned opposite the light-emitting and receiving nozzle.

15. A quality measuring device as described in claim 9, wherein the container of the light absorption section has a container-side light-transmitting section, and the container-side light-transmitting section is positioned so as to face the end of the optical fiber for transmitted light.

16. A quality measuring device according to claim 1, wherein the target substance is a nucleic acid or a protein.

17. A quality measuring device, wherein the fluorescence unit is a PCR unit that performs a PCR reaction of the nucleic acid.

18. A quality measuring device as described in claim 1, wherein the control unit performs qualitative and / or quantitative processing of the target substance using the measured fluorescence, chemiluminescence, and / or absorbance.

19. A quality measuring device as described in claim 18, further comprising an imaging unit that images an electrophoretic pattern obtained from the electrophoresis, and the control unit executes an evaluation process for the quality of the target substance based on the electrophoretic pattern.

20. A sample processing system comprising: a sample processing device that performs a process for extracting and purifying a target substance (e.g., nucleic acid) from a sample; and the quality measuring device according to claim 1.

21. A sample processing system according to claim 20, comprising a moving device for moving the target substance solution from the sample processing device to the quality measuring device.

22. A sequencer that performs sequencing of nucleic acid obtained from a specimen, comprising: a specimen processing device having a pre-treatment section that extracts and purifies the nucleic acid from the specimen, a pre-treatment side PCR section that amplifies the nucleic acid by PCR using the nucleic acid solution obtained from the pre-treatment section, and a first nozzle unit that moves the nucleic acid solution; a quality measuring device as described in claim 1 that measures the quality of the nucleic acid using the nucleic acid solution obtained from the pre-treatment section; a transfer device that transfers the nucleic acid solution from the pre-treatment section to the quality measuring device; a flow cell section that performs sequencing of the nucleic acid using the nucleic acid solution; and a control section that controls processing in the specimen processing device, the quality measuring device, the transfer device, and the flow cell section, and which performs sequencing of the nucleic acid when the control section determines that the quality of the nucleic acid measured by the quality measuring device satisfies at least one predetermined condition.

23. A sequencer as described in claim 22, wherein the pre-treatment section comprises a nucleic acid extraction section for extracting nucleic acid from the sample, a sizing section for sizing the extracted nucleic acid, and an adapter binding section for binding adapters to both ends of the sized nucleic acid.

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