Channel substrate, cartridge, determination system, and determination method

Abstract

The present invention is provided with: a channel through which a fluid mixture containing a sample and a labeling substance that reacts with a detection target contained in the sample flows; and a standard part that is positioned in a region different from that of the channel and is to be compared with the fluid mixture.

Description

Flow path substrate, cartridge, determination system and determination method

[0001] The present disclosure relates to a flow path substrate and the like.

[0002] The dual-chamber reaction vessel of Patent Document 1 includes a first chamber for receiving a liquid sample, a second chamber physically independent from the first chamber, and a channel connecting the first chamber to the second chamber. The first chamber contains amplification reagents, and the second chamber contains an enzyme.

[0003] Japanese Patent Publication No. 11-4678

[0004] A flow path substrate according to one aspect of the present disclosure includes a flow path in which a mixed fluid containing a sample and a labeled substance that reacts with a target substance contained in the sample is located, and a standard section located in a region different from the flow path and for comparison with the mixed fluid.

[0005] Furthermore, a cartridge according to one aspect of the present disclosure includes a flow path substrate having a flow path inside which a mixed fluid containing a sample and a labeled substance that reacts with the target substance contained in the sample is located, and a housing that stores the flow path substrate, and the housing includes a standard portion for comparison with the mixed fluid.

[0006] Furthermore, a determination system according to one aspect of the present disclosure includes a flow path substrate having a flow path inside which a mixed fluid containing a sample and a labeled substance that reacts with the target substance contained in the sample is located, and a standard section located in an area different from the flow path and for comparison with the mixed fluid, and a first determination section that determines the presence or absence of the target substance using optical information from the mixed fluid and the standard section.

[0007] Furthermore, a determination system according to one aspect of the present disclosure includes a flow path substrate having a flow path inside which a mixed fluid containing a sample and a labeled substance that reacts with the target substance contained in the sample is located, a housing that stores the flow path substrate and has a standard section that is compared with the mixed fluid, and a first determination section that determines the presence or absence of the target substance using optical information from the mixed fluid and the standard section.

[0008] Furthermore, a determination method according to one aspect of the present disclosure includes a flow path substrate having a mixed fluid containing a sample and a labeled substance that reacts with the target substance contained in the sample located therein, and a standard portion located in an area different from the flow path and for comparison with the mixed fluid, the method including the steps of measuring optical information of the mixed fluid and the standard portion, and determining the presence or absence of the target substance using the optical information of the mixed fluid and the standard portion.

[0009] Furthermore, a determination method according to one aspect of the present disclosure includes the steps of measuring optical information of a mixed fluid containing a sample and a labeled substance that reacts with a target substance contained in the sample in a flow path substrate having a flow path inside which the mixed fluid is located, and a standard portion in a housing that stores the flow path substrate and has a standard portion for comparison with the mixed fluid, and determining the presence or absence of the target substance using the optical information of the mixed fluid and the standard portion.

[0010] 1 is a schematic diagram showing an example of a detection system of the present disclosure; FIG. 1 is a schematic diagram showing an example of a configuration of a cartridge of the present disclosure, and a schematic diagram of a window portion of the cartridge of the present disclosure when viewed from above; FIG. 2 is a schematic diagram for explaining an example of use of the cartridge and detection device of the present disclosure; FIG. 3 is a block diagram showing an example of a detection system of the present disclosure; FIG. 4 is a flowchart showing an example of a processing flow of a determination method of the present disclosure; FIG. 5 is a plan view showing an example of a flow path substrate of a cartridge of the present disclosure; FIG. 6 is a schematic diagram showing an example of a first region and a second region arranged in a flow path provided in a flow path substrate according to a first embodiment of the present disclosure; FIG. 7 is a plan view showing another example of a flow path substrate of a cartridge of the present disclosure; FIG. 8 is a diagram for explaining a method of specifying the position of a storage portion using an internal standard;

[0011] An embodiment of the present disclosure can provide a flow path substrate, a cartridge, a determination system, or a determination method that can facilitate detection of a detection target.

[0012] [Detection System] Fig. 1 is a schematic diagram showing an example of a detection system 1 according to the present disclosure. Fig. 1 is a schematic diagram showing an example of the appearance of a cartridge 2 and an example of the appearance of a detection device 3. Fig. 2 is a block diagram showing an example of the detection system 1. As shown in Fig. 1, the detection system 1 may include the cartridge 2 and the detection device 3. In this specification, the detection system is also referred to as a determination system.

[0013] The cartridge 2 is a test kit for detecting a target substance contained in a specimen collected from a subject. The cartridge 2 may be a test kit capable of amplifying nucleic acid derived from the target substance using a reagent when the target substance is contained in a specimen inserted into the cartridge 2. Various pathogens may be the target substance. For example, SARS-CoV-2, influenza virus, hemolytic streptococcus, Mycoplasma pominis, and respiratory syncytial virus (RS virus) may be the target pathogens to be tested. The cartridge 2 may include a main body portion 21 and a bottle portion 22.

[0014] The subject is not limited to a human, but may be any living organism that can harbor a virus or bacterium, such as a mammal, bird, reptile, or amphibian.

[0015] The specimen may be saliva, urine, sweat, nasal mucus, blood, cells, etc. collected from the body of a subject, or may be soil and water collected from an object to be inspected, such as the ground, a river, the sea, etc. Alternatively, the specimen may be an attachment collected from the surface of an object to be inspected, such as a handrail, a door, clothing, shoes, or a toilet bowl.

[0016] The detection target may be, for example, a virus or a bacterium. The type of virus or bacterium contained in the sample is not limited to one type, and may be two or more types. When the detection target is a virus, examples of the type of virus include influenza virus, coronavirus (e.g., SARS-CoV-2), respiratory syncytial virus (RS virus), human metapneumovirus, norovirus, HIV (human immunodeficiency virus), herpes virus, streptococcus, and Mycoplasma pominis.

[0017] A specimen collected from a subject may be contained in the bottle portion 22. The main body portion 21 may include a flow path substrate 4 (see FIG. 6 ) that receives a liquid containing the specimen contained in the bottle portion 22 and, when the specimen contains a target substance, contains a reagent that reacts with nucleic acid derived from the target substance. The nucleic acid derived from the target substance may be extracted in the bottle portion 22, as will be described in detail below.

[0018] The detection device 3 is a device that detects nucleic acid derived from a detection target when the nucleic acid is amplified in the cartridge 2. The detection device 3 may have a housing that can receive the cartridge 2.

[0019] The detection device 3 may determine whether nucleic acid derived from the detection target is present at a certain concentration or above. The detection device 3 may determine that the sample is positive when nucleic acid derived from the detection target is present at a certain concentration or above. Alternatively, the detection device 3 may be a measurement device that measures the concentration of nucleic acid derived from the detection target. In this embodiment, the detection device 3 is described as determining whether the sample is positive. However, a determination device separate from the detection device 3 may determine whether the sample is positive by obtaining data corresponding to the amount of nucleic acid detected by the detection device 3 from the detection device 3. The specific configurations of the cartridge 2 and the detection device 3 will be described later.

[0020] [NASBA Method] In the flow path substrate 4, nucleic acid derived from the detection target may be amplified by an isothermal nucleic acid amplification method. Examples of the isothermal nucleic acid amplification method include NASBA (Nucieic Acid Sequence-Based Amplification). Other examples include NEAR (Nicking Enzyme Amplification), LAMP (Loop-mediated Isothermal Amplification), and TMA (Transcription-mediated Amplification). In this embodiment, NASBA may be used as the isothermal nucleic acid amplification method.

[0021] The NASBA method is an isothermal nucleic acid amplification method that uses three types of enzymes (AMV reverse transcriptase, RNase H, and T7 RNA polymerase) and two types of primers. In the NASBA method, simply adding a NASBA substrate and enzyme to the template RNA results in amplified antisense single-stranded RNA. Because the entire series of steps in the NASBA reaction proceeds isothermally, RNA nucleic acid amplification can be performed without complex temperature control. Furthermore, because the amplification product of the NASBA reaction is single-stranded RNA, sequence-specific detection can be performed using a detection probe without a denaturation step.

[0022] [Specific Configuration of Cartridge] Reference numeral 1101 in Fig. 2 is a schematic diagram showing an example of the configuration of the cartridge 2. Reference numeral 1102 in Fig. 2 is a schematic diagram of the window portion 211 of the cartridge 2 as viewed from above. Fig. 3 is a schematic diagram for explaining an example of how the cartridge 2 and the detection device 3 are used.

[0023] As indicated by reference numeral 1101 in FIG. 2 , the main body 21 may include a flow path substrate 4. In other words, the cartridge 2 can be expressed as including a housing of the main body 21 that houses the flow path substrate 4. The flow path substrate 4 may include a liquid receiving portion 41 that receives a liquid. Furthermore, as indicated by reference numeral 1102 in FIG. 2 , the flow path substrate 4 may include a storage portion 46 that stores a liquid that has flowed through the flow path substrate 4. Furthermore, the flow path substrate 4 may include a flow path 45 through which a mixed fluid containing a sample and a labeling substance that reacts with the target substance contained in the sample flows (see FIG. 6 ). In other words, the mixed fluid is located inside the flow path 45. The housing of the main body 21 may be made of an opaque material. In this case, the main body 21 may include a window portion 211 so that the interior of the main body 21 can be viewed.

[0024] The bottle portion 22 is a container capable of holding a liquid. The bottle portion 22 may be made of a transparent material, but may also be made of an opaque material.

[0025] As indicated by the arrow at reference numeral 1101 in Fig. 2 , the liquid receiving section 41 receives the liquid that has flowed in from the bottle section 22. The liquid received by the liquid receiving section 41 flows through a flow path 45 (see Fig. 6 ) arranged in the flow path substrate 4, and is then stored in a storage section 46. A reagent may be arranged in the flow path 45. Furthermore, as indicated by reference numeral 1102 in Fig. 2 , an internal standard (standard section) 47 may be provided near the storage section 46. The internal standard 47 may be used for optical comparison with the storage section 46.

[0026] The internal standard 47 may be located in a region different from the flow channel 45 and used for comparison with a mixed fluid containing a sample and a labeling substance that reacts with the target substance contained in the sample.

[0027] For example, the internal standard 47 may be located in a region different from the flow channel 45 and used for comparison with the fluorescence intensity emitted by the mixed fluid. Here, the internal standard 47 may be used for comparison with the mixed fluid located in the flow channel 45. The internal standard 47 may be used for comparison with the mixed fluid located in the reservoir 46.

[0028] 2 , the main body 21 is formed with a window 211 so that, when viewed from above, the reservoirs 46 and internal standards 47 arranged on the flow path substrate 4 can be seen through the window 211. When the flow path substrate 4 includes a plurality of reservoirs 46 and a plurality of internal standards 47, the window 211 may be formed so that all of the reservoirs 46 and all of the internal standards 47 can be seen.

[0029] That is, the reservoir 46 and the internal standard 47 are optically exposed in the flow path substrate 4 included in the cartridge 2. In other words, the reservoir 46 and the internal standard 47 may be disposed in the main body 21 so as to be optically exposed from the window 211.

[0030] The labeling substance may be a substance that specifically reacts with the target substance. Alternatively, the labeling substance may be a substance whose properties change upon reaction with the target substance. In other words, the labeling substance may be a substance for detecting the target substance.

[0031] The labeling substance may be optically observable, and may include, but is not limited to, a dye, a luminescent substance, or a fluorescent substance. The labeling substance may have optical properties that change upon reaction with the target of detection. For example, the labeling substance may include a fluorescent substance. When the labeling substance includes a fluorescent substance, the labeling substance bound to the nucleic acid emits fluorescence having a specific wavelength when excitation light is irradiated onto the storage section 46. When the labeling substance includes a fluorescent substance, the fluorescence of the labeling substance may increase upon reaction with the target of detection. A labeling substance that specifically binds to the nucleic acid amplified in the channel 45 may be disposed in the channel 45 of the channel substrate 4.

[0032] Examples of fluorescent substances contained in labeling substances include 6-caroxyfluorescein (FAM), Texas Red (TR) (registered trademark), and cyanine (CY)-based materials. When the fluorescent substance is 6-FAM, it emits fluorescence with a peak wavelength of 517 nm when irradiated with excitation light with a peak wavelength of 494 nm. When the fluorescent substance is Texas Red, it emits fluorescence with a peak wavelength of 615 nm when irradiated with excitation light with a peak wavelength of 596 nm. When the fluorescent substance is Cy3-carboxylic acid, it emits fluorescence with a peak wavelength of 570 nm when irradiated with excitation light with a peak wavelength of 555 nm.

[0033] The labeling substance may be, for example, a molecular beacon. A molecular beacon is a type of probe DNA. Each molecule of the probe DNA is modified with a fluorescent molecule and a quencher molecule. The quencher molecule is configured to absorb light in a wavelength band corresponding to the fluorescence wavelength of the fluorescent molecule, for example. The distance between the fluorescent molecule and the quencher molecule in the probe DNA that is not bound to the enzymatically amplified nucleic acid molecule is closer than the distance between the fluorescent molecule and the quencher molecule in the probe DNA that is bound to the enzymatically amplified nucleic acid molecule. The base sequence of the probe DNA may be appropriately designed based on the base sequence of the nucleic acid to be detected.

[0034] The internal standard 47 may be any optically observable material, and may include, but is not limited to, a dye, a luminescent material, or a fluorescent material. The internal standard 47 may also include a fluorescent material. The internal standard 47 may also include a fluorescent material that is excited by an excitation wavelength that excites the fluorescent material contained in the labeling material. The fluorescent material contained in the internal standard 47 and the fluorescent material contained in the labeling material may be the same type of fluorescent material.

[0035] As shown by reference numeral 1112 in FIG. 3 , the sampler 23 may be attached to the main body 21. The sampler 23 may include a specimen collection section 231 and a fixing section 232. The specimen collection section 231 is a section for collecting a specimen. The specimen collection section 231 may be, for example, a pleated resin member. The fixing section 232 may fix the bottle section 22 to the main body 21 when the sampler 23 is inserted into the bottle section 22. The connection section between the main body 21 and the bottle section 22 may have, for example, a screw structure. Furthermore, a lid may be provided at the connection section of the bottle section 22 with the main body 21. This allows the interior of the bottle section 22 to be sealed. The lid may be made of aluminum. The subject may remove the lid and insert the sampler 23 into the bottle section 22.

[0036] As indicated by reference numeral 1111 in Fig. 3 , when collecting saliva as a specimen, for example, the subject inserts the specimen collection portion 231 into the subject's mouth to cause saliva to adhere to the specimen collection portion 231. As indicated by reference numeral 1112 in Fig. 3 , the subject applies saliva to the specimen collection portion 231 and then inserts the sampler 23 into the bottle portion 22.

[0037] The bottle portion 22 may contain a buffer solution 24. An example of the buffer solution 24 is a NASBA solution. The buffer solution 24 may contain, for example, a surfactant and an RNA (ribonucleic acid) degrading enzyme inhibitor. However, the RNA degrading enzyme inhibitor may be disposed on the flow path substrate 4. In this case, the buffer solution 24 may not contain an RNA degrading enzyme inhibitor. An example of a surfactant is Tween-20 (polysorbate 20), which is an example of a nonionic surfactant. When the buffer solution 24 contains a nonionic surfactant, nucleic acids derived from the detection target can be extracted in the bottle portion 22 by mixing the sample and the buffer solution 24.

[0038] When the subject inserts the sampler 23 into the bottle portion 22, the saliva adhering to the specimen collection portion 231 is mixed with the buffer solution 24. The buffer solution 24 mixed with the specimen may be an example of a liquid that the bottle portion 22 can contain.

[0039] As indicated by reference numeral 1113 in Fig. 3 , the detection device 3 may include a receiving section 36 that can receive the cartridge 2. As indicated by reference numeral 1113 in Fig. 3 , the subject attaches the cartridge 2 containing the sample to the receiving section 36, and then turns the detection device 3 upside down, as indicated by reference numeral 1114 in Fig. 3 . As a result, the buffer solution 24 mixed with the sample flows, due to its own weight, down the sampler 23 and into the flow path substrate 4 provided in the main body section 21. The buffer solution 24 mixed with the sample may be an example of a liquid that the liquid receiving section 41 receives.

[0040] As a negative control, only the buffer solution 24 may be poured into the flow path substrate 4. In this case, the buffer solution 24 itself is an example of a liquid that can be accommodated in the bottle portion 22, and an example of a liquid that can be received by the liquid receiving portion 41. Also, a substance that contains a pathogen other than the detection target may be mixed into the buffer solution 24. In this case, the buffer solution 24 mixed with the substance is an example of a liquid that can be accommodated in the bottle portion 22, and an example of a liquid that can be received by the liquid receiving portion 41.

[0041] 4 is a block diagram showing an example of the detection system 1. As shown in Fig. 4, the detection device 3 may be a device that detects nucleic acid amplified in a flow channel 45 provided in a flow channel substrate 4. The detection device 3 may include, for example, a heating unit 31, a pressurizing unit 32, a light irradiating unit (excitation light source) 33, an imaging unit 34, and a control unit 35.

[0042] The heating unit 31 may be a member that heats the cartridge 2 inserted into the detection device 3. The heating unit 31 may heat the cartridge 2 to a temperature that promotes the reaction between the nucleic acid and the reagent in the flow path substrate 4. When amplifying nucleic acids by the NASBA method, the heating unit 31 may heat the cartridge 2 to a temperature of 37°C to 41°C. In this embodiment, the heating unit 31 first heats the cartridge 2 at a temperature of 80°C to 95°C for 3 to 10 minutes. This disrupts pathogens and liberates nucleic acid components from the pathogens. It also inactivates DNA (deoxyribonucleic acid) degrading enzymes. Thereafter, the heating unit 31 may maintain the cartridge 2 at a temperature of 37°C to 41°C.

[0043] The heating unit 31 may heat different parts of the cartridge 2 at different temperatures. For example, the heating unit 31 may have a first heating unit that heats the bottle unit 22 to a temperature of 80°C to 95°C, and a second heating unit that heats the main body unit 21 to a temperature of 37°C to 41°C. This allows pathogens to be crushed in the bottle unit 22, liberating nucleic acid components from the pathogens. Furthermore, the DNase can be inactivated in the bottle unit 22. The liquid in the bottle unit 22 then flows into the flow path substrate 4 provided in the main body unit 21, which is maintained at a temperature of 37°C to 41°C, and nucleic acid amplification may be performed in the flow path substrate 4.

[0044] The temperature to which the heating unit 31 heats may be the set temperature of the heating unit 31. Alternatively, it may be the liquid temperature of the liquid whose temperature is to be adjusted by the heating unit 31. When the temperature to which the heating unit 31 heats is the liquid temperature of the liquid whose temperature is to be adjusted by the heating unit 31, for example, the average temperature or central temperature of the liquid may be used. Here, the average temperature may be, for example, the average liquid temperature over a predetermined period of time. The central temperature may be, for example, the temperature at the center between the highest and lowest liquid temperatures. The central temperature may be calculated as the sum of the highest and lowest temperatures divided by 2. The detection device 3 may be equipped with a temperature sensor that detects the liquid temperature of the liquid whose temperature is to be adjusted by the heating unit 31. The heating unit 31 may heat the liquid whose temperature is to be detected by the temperature sensor so as to maintain the liquid temperature detected by the temperature sensor.

[0045] The pressurizing unit 32 may be a member that pressurizes the bottle portion 22 of the cartridge 2 inserted into the detection device 3. When the detection device 3 is configured to pressurize the bottle portion 22 by the pressurizing unit 32, the bottle portion 22 may be made of a material that is deformed by the pressure of the pressurizing unit 32. The bottle portion 22 is deformed by the pressure of the pressurizing unit 32, which makes it easier for the liquid in the bottle portion 22 to flow into the main body portion 21.

[0046] The pressurizing unit 32 may be a member that applies pressure to at least a portion of the side surface of the bottle part 22. The pressurizing unit 32 may be a member that clamps the side surface of the bottle part 22. The pressurizing unit 32 may be disposed in the detection device 3 at a position where it can apply pressure to the side surface on the bottom side of the bottle part 22, for example.

[0047] The light irradiating unit 33 may be a light source that irradiates the storage unit 46 and the internal standard 47 of the flow path substrate 4 with light through the window 211 of the main body 21. The light irradiating unit 33 irradiates, for example, excitation light onto the storage unit 46 and the internal standard 47.

[0048] The imaging unit 34 is a member that images the storage unit 46 and the internal standard 47 through the window 211 of the main body 21. When the storage unit 46 and the internal standard 47 are arranged in the main body 21 so as to be optically exposed from the window 211, the imaging unit 34 images the storage unit 46 and the internal standard 47 through the window 211 of the main body 21. The imaging unit 34 may be any unit that can obtain an image, and may take a still image as the image, or may take a video as the image.

[0049] The control unit 35 may comprehensively control each component included in the detection device 3. The control unit 35 may include, for example, a position determination unit (second determination unit) 350, an intensity measurement unit 351, and a positive determination unit (first determination unit) 352.

[0050] The control unit 35 may first control the light irradiating unit 33 to irradiate the storage unit 46 and the internal standard 47 with light. In this state, the control unit 35 may control the imaging unit 34 to capture an image including the storage unit 46 and the internal standard 47. For example, the control unit 35 controls the light irradiating unit 33 to irradiate the storage unit 46 and the internal standard 47 with excitation light, and then analyzes the image captured by the imaging unit 34 to obtain brightness. In this way, the control unit 35 may measure the intensities of fluorescence emitted by the substances contained in the storage unit 46 and the internal standard 47, respectively. The control unit 35 may also control the imaging unit 34 to capture an image including the storage unit 46 and the internal standard 47 without irradiating light.

[0051] The imaging unit 34 captures an image of the internal standard 47 to obtain a captured image having optical information measured from the internal standard 47. The position determination unit 350 may identify the position of the internal standard 47 in the captured image. The optical information may be, for example, information obtained from a dye, a luminescent substance, or a fluorescent substance. The optical information may be, for example, information such as a wavelength luminance value. For example, the position determination unit 350 may identify the position of the internal standard 47 in the captured image based on the fluorescence emitted from the internal standard 47.

[0052] The position determination unit 350 may determine whether the position of the image of the fluorescence emitted by the internal standard 47 in the captured image is within a normal range. For example, the position determination unit 350 may make this determination based on whether or not light emission with an intensity equal to or greater than a threshold is detected within a predetermined range in the captured image. Furthermore, when there are multiple internal standards 47, the position determination unit 350 may determine whether the positions of the images of the fluorescence emitted by the multiple internal standards 47 are within the normal range.

[0053] If the position determination unit 350 does not detect light emission with an intensity equal to or greater than the threshold within a predetermined range in the captured image, the position determination unit 350 may determine that the captured image was not acquired normally. If the position determination unit 350 determines that the captured image was not acquired normally, the control unit 35 may notify the user that an abnormality has occurred via an output device such as a display unit and / or a speaker (not shown), and then terminate the process of determining the presence or absence of a detection target.

[0054] The position determination unit 350 may determine whether fluorescence of a predetermined intensity or greater is received from the internal standard 47. The position determination unit 350 may determine that the image capture unit 34 is operating normally when fluorescence of a predetermined intensity or greater is received in a predetermined range in the captured image. The determination of whether optical information of a predetermined value or greater is measured may be performed once after starting the detection device 3 and before measuring optical information obtained from the sample. For example, the determination of whether fluorescence of a predetermined intensity or greater is received, i.e., the measurement of the predetermined fluorescence intensity, may be performed once after starting the detection device 3 and before measuring the intensity of fluorescence emitted by the sample.

[0055] Furthermore, the position determination unit 350 may identify the position of the internal standard 47 in the captured image, and identify the position of the storage unit 46 in the captured image from the identified position of the internal standard 47. When there are multiple storage units 46, the position determination unit 350 may identify the position of each storage unit 46 in the captured image from the identified position of the internal standard 47. The position determination unit 350 may identify the position of the flow path 45 in the captured image based on the identified position of the internal standard 47.

[0056] The intensity measurement unit 351 may measure optical information of the storage unit 46 and the internal standard 47. The optical information of the storage unit 46 may be optical information of a mixed fluid containing a sample and a labeling substance that reacts with the target substance contained in the sample. The optical information may be, for example, information obtained from a dye, a luminescent substance, or a fluorescent substance. The optical information may be, for example, information such as wavelength luminance values. For example, the intensity measurement unit 351 may analyze an image captured by the imaging unit 34 and measure luminance as optical information obtained from the substance contained in the storage unit 46 and the internal standard 47. For example, the intensity measurement unit 351 may measure the intensity of fluorescence emitted by the substance contained in the storage unit 46 and the internal standard 47 when exposed to excitation light.

[0057] The intensity measurement unit 351 may refer to the positions of the storage unit 46 and the internal standard 47 identified by the position determination unit 350. The intensity measurement unit 351 may refer to the positions of the storage unit 46 and the internal standard 47 identified by the position determination unit 350, and measure the optical information of the storage unit 46 and the internal standard 47 in association with each other.

[0058] The intensity measurement unit 351 may measure optical information obtained from the reservoir 46 connected to the flow channel 45 in which the reagent is placed as optical information of the detection target. For example, the intensity measurement unit 351 may measure the intensity of fluorescence emitted from the reservoir 46 connected to the flow channel 45 in which the reagent is placed as the intensity of fluorescence of the detection target.

[0059] Furthermore, the intensity measurement unit 351 may measure optical information emitted from the reservoir 46 connected to the negative control flow path 45 as optical information of the negative control. For example, the intensity measurement unit 351 may measure the intensity of fluorescence emitted from the reservoir 46 connected to the negative control flow path 45 as the intensity of fluorescence of the negative control. The negative control flow path 45 will be described later. The position of the flow path 45 in which the reagent is placed and the position of the negative control flow path 45 may be determined in advance.

[0060] The positive determination unit 352 may determine whether a certain concentration or more of nucleic acid derived from the detection target is present in the sample contained in the liquid stored in the storage unit 46 based on the optical information obtained from the storage unit 46 measured by the intensity measurement unit 351. For example, the positive determination unit 352 may determine whether a certain concentration or more of nucleic acid derived from the detection target is present in the sample contained in the liquid stored in the storage unit 46 based on the intensity of fluorescence emitted from the storage unit 46 measured by the intensity measurement unit 351. The positive determination unit 352 may determine that the sample is positive if it determines that a certain concentration or more of nucleic acid derived from the detection target is present. That is, the positive determination unit 352 may determine the presence or absence of the detection target using optical information from the labeling substance and internal standard 47 stored in the storage unit 46. The positive determination unit 352 may determine the presence or absence of the detection target using optical information from the internal standard 47 and optical information from a mixed fluid containing the sample stored in the storage unit 46 and a labeling substance that reacts with the detection target contained in the sample. For example, the positive determination unit 352 may determine the presence or absence of the target substance using the intensity of fluorescence emitted by the labeled substance and the internal standard 47 stored in the storage unit 46. That is, the internal standard 47 may be used for comparison with the labeled substance. The internal standard 47 may be used for comparison with the above-mentioned mixed fluid. Comparing the internal standard 47 with the labeled substance can improve the reproducibility of detection of the target substance. Furthermore, the positive determination unit 352 may determine the presence or absence of the target substance for each of the multiple storage units 46 that store the above-mentioned mixed fluid and are provided in the flow path substrate 4. This makes it possible to determine the presence or absence of multiple target substances at once.

[0061] For example, the positive determination unit 352 may determine whether a certain concentration or more of nucleic acid derived from the detection target is present by comparing the value indicated by the optical information obtained from the storage unit 46 with a threshold value. For example, the threshold value may be optical information associated with the minimum concentration of nucleic acid derived from the detection target that should be determined as positive. The threshold value may be set in advance through experiments, etc. For example, the positive determination unit 352 may determine whether a certain concentration or more of nucleic acid derived from the detection target is present by comparing the intensity of fluorescence emitted from the storage unit 46 with the threshold value.

[0062] The positive determination unit 352 may calculate a difference value by subtracting the value indicated by the optical information of the negative control from the value indicated by the optical information of the detection target. For example, the positive determination unit 352 may calculate a difference value by subtracting the fluorescence intensity of the negative control from the fluorescence intensity of the detection target. The positive determination unit 352 may compare the difference value with a threshold, and determine that the sample is positive if the difference value is equal to or greater than the threshold, and determine that the sample is negative if the difference value is less than the threshold.

[0063] Alternatively, the positive determination unit 352 may determine whether the sample is positive by comparing the difference value with the value indicated by the optical information obtained from the internal standard 47. The optical information of at least two of the multiple internal standards 47 included in the flow path substrate 4 may be different from each other. For example, two internal standards 47 having different concentrations of fluorescent material may be disposed. For example, the amounts of fluorescent material contained in at least two of the multiple internal standards 47 included in the flow path substrate 4 may be different from each other. The positive determination unit 352 may determine the presence or absence of the target substance based on the optical information of the internal standards 47 having different optical information. For example, the positive determination unit 352 may determine the presence or absence of the target substance based on the intensity of fluorescence emitted by the internal standards 47 having different amounts of fluorescent material. This can improve the reproducibility of the detection of the target substance. The positive determination unit 352 may determine the sample is positive if the difference value is within the range of values ​​indicated by the optical information obtained from the two internal standards 47. The optical information obtained from the two internal standards 47 may be optical information associated with the concentration of nucleic acid derived from the detection target to be determined as positive. The optical information obtained from the two internal standards 47 may be set in advance by an experiment or the like.

[0064] For example, the positive determination unit 352 may determine whether the sample is positive by comparing the difference value with the intensity of the fluorescence emitted from the internal standard 47. The positive determination unit 352 may determine that the sample is positive if the difference value is within the range of the fluorescence intensities indicated by the two internal standards 47.

[0065] The control unit 35 may also create a calibration curve based on optical information obtained from the multiple internal standards 47. For example, the control unit 35 may create a calibration curve based on the intensities of fluorescence emitted by the multiple internal standards 47. In this case, the control unit 35 may use the calibration curve to calculate the concentration of nucleic acid corresponding to the above-mentioned difference value. The positive determination unit 352 may determine that the sample is positive if the calculated nucleic acid concentration is equal to or greater than a reference value. The reference value may be a value associated with the concentration of nucleic acid derived from the detection target that should be determined to be positive. The reference value may be set in advance through experiments, etc. This can improve the reproducibility of detection of the detection target.

[0066] In the following explanation, as an example, the labeling substance and internal standard 47 are fluorescent substances, and the control unit 35 measures the intensity of fluorescence as the optical information measured from the labeling substance and internal standard 47.

[0067] Furthermore, when the detection system 1 of the present disclosure is applied to infectious disease testing, the amount of pathogens such as viruses and bacteria may be calculated from the concentration of nucleic acids calculated using the calibration curve described above.

[0068] [Determination Process Flow] FIG. 5 is a flowchart illustrating an example of the process flow for determining the presence or absence of a target substance performed by the detection system 1. As shown in FIG. 5, the control unit 35 first instructs the light irradiation unit 33 to irradiate the mixed fluid and the internal standard 47 in the flow path substrate 4 with excitation light (step S11). The mixed fluid may contain a sample and a labeling substance that reacts with the target substance contained in the sample. Next, the position determination unit 350 determines whether the position of the fluorescence image emitted by the internal standard 47 in the captured image is within a normal range (step S12). Next, the intensity measurement unit 351 measures optical information of the mixed fluid and the internal standard 47. For example, the intensity measurement unit 351 measures the intensity of the fluorescence emitted by the mixed fluid and the internal standard 47 (step S13). Next, the positive determination unit 352 determines the presence or absence of a target substance using optical information of the mixed fluid stored in the storage unit 46 and the internal standard disposed in the internal standard 47. For example, the positive determination unit 352 determines the presence or absence of the detection target using the intensity of fluorescence emitted by the mixed fluid stored in the storage unit 46 and the internal standard disposed in the internal standard 47 (step S14). Then, the process ends. If the mixed fluid and the internal standard 47 exhibit (reflect or emit) light without requiring excitation light, step S11 may be omitted.

[0069] [First Form of Flow Channel Substrate] Fig. 6 is a plan view showing an example of a flow channel substrate 4. The flow channel substrate 4 is an example of a flow channel substrate 4. Fig. 7 is a schematic diagram showing an example of a first region 451 and a second region 452 arranged in a flow channel 45 provided in the flow channel substrate 4.

[0070] 6, the flow path substrate 4 may include a liquid receiving section 41 that receives a liquid, and a flow path 45 that connects to the liquid receiving section 41. The flow path 45 may have a first region 451 in which a first reagent 101 containing a primer is disposed, and a second region 452 in which a second reagent 102 containing an enzyme that amplifies nucleic acid is disposed at a position different from the first region 451. In the flow path 45 that communicates with the liquid receiving section 41, the space in which the first region 451 is located and the space in which the second region 452 is located may communicate with each other.

[0071] In this embodiment, as shown in FIG. 6 , the flow path substrate 4 may include a liquid receiving section 41, a branch flow path 42, a flow path 45, a storage section 46, and an internal standard 47. There may be one or more flow paths 45. There may also be one or more internal standards 47. In this embodiment, the flow path substrate 4 will be described as including four flow paths 45A to 45D and four storage sections 46A to 46D, but this is not limiting. In addition, the flow path substrate 4 will be described as including five internal standards 47, but this is not limiting.

[0072] 4, the liquid receiving portion 41 is located above the flow path 45 and the storage portion 46. Therefore, the liquid flowing from the bottle portion 22 due to its own weight easily flows from the liquid receiving portion 41 to the storage portion 46 via the flow path 45.

[0073] Liquid receiving section 41 may be an opening that receives the liquid flowing from bottle section 22. That is, liquid receiving section 41 is for receiving a sample and may be connected to flow path 45. Branch flow path 42 may be a flow path that connects liquid receiving section 41 and each of flow paths 45A to 45D. That is, flow paths 45A to 45D may branch from branch flow path 42 that is connected to liquid receiving section 41. Branch flow path 42 may be in communication with liquid receiving section 41 and each of flow paths 45A to 45D.

[0074] The flow path substrate 4 may include a filter section 420. The filter section 420 may be a section whose flow path width is narrower than other sections. The filter section 420 may be located on the opposite side of the reservoir section 46 from the position of the reagent disposed in the flow path 45. That is, the filter section 420 may be located upstream of the position of the reagent disposed in the flow path 45. When solid impurities are contained in the liquid flowing from the liquid receiving section 41, the possibility of the impurities flowing downstream of the filter section 420 can be reduced. Furthermore, the filter section 420 may be located upstream of the branching positions of the flow paths 45A to 45D. As shown in FIG. 6 , the branch flow path 42 may include the filter section 420. This reduces the possibility of the impurities flowing into the flow paths 45A to 45D when solid impurities are contained in the liquid flowing from the liquid receiving section 41.

[0075] However, the flow path substrate 4 does not have to include the branch flow path 42. In this case, the flow paths 45A to 45D may branch from the liquid receiving section 41. When there is only one flow path 45, the flow path 45 may be connected to the liquid receiving section 41. Furthermore, the filter section may be located in each of the flow paths 45A to 45D on the opposite side of the reservoir section 46 from the position of the reagent placed in the flow path 45. Each of the flow paths 45A to 45D may have a filter section upstream of the position where the first reagent 101 is placed or the position where the first reagent 101 can be placed.

[0076] 6, all of the flow paths 45A to 45D may branch off from one liquid receiving section 41 or from a branch flow path 42 connected to one liquid receiving section 41. However, the flow path substrate 4 may be provided with a plurality of liquid receiving sections 41. In this case, each flow path 45 may be connected to each liquid receiving section 41, or one or more flow paths 45 may be connected to each liquid receiving section 41.

[0077] The flow paths 45A to 45D may be flow paths for flowing a liquid. A mixed fluid containing a specimen and a labeling substance that reacts with the target substance contained in the specimen may be flowed through at least one of the flow paths 45A to 45D. Each of the flow paths 45A to 45D may be a flow path that receives a liquid flowing from the liquid receiving section 41 and flows the liquid to the reservoirs 46A to 46D connected to the flow paths 45A to 45D, respectively. Each of the flow paths 45A to 45D may be in communication from the liquid receiving section 41 to the reservoirs 46A to 46D.

[0078] In this embodiment, the flow paths 45A to 45D each have a filter section 450A to 450D upstream of the branching position and upstream of the position where the first reagent 101 is placed or the position where the first reagent 101 can be placed.

[0079] In at least one of the flow paths 45A to 45D, the reagent may be disposed on the opposite side of the branching position of the flow paths 45A to 45D from the liquid-receiving section 41. In this way, when the liquid received in the liquid-receiving section 41 contains nucleic acid derived from the detection target, the nucleic acid derived from the detection target can be amplified in the flow path 45.

[0080] In this embodiment, one of the flow paths 45A to 45D may have a first region 451 in which a first reagent 101 is disposed, and a second region 452 in which a second reagent 102 is disposed at a position different from the first region 451.

[0081] The liquid received in liquid receiving section 41 flows through each of flow paths 45A to 45D and is stored in reservoirs 46A to 45D. In this embodiment, as shown in FIG. 6 , flow path 45B has a first region 451 and a second region 452. Therefore, the liquid flowing through flow path 45B comes into contact with first reagent 101 and second reagent 102. If the sample contains a detection target, nucleic acid derived from the detection target can be amplified by first reagent 101 and second reagent 102.

[0082] Therefore, nucleic acid derived from the detection target can be amplified by a simple operation of flowing a liquid through the flow channel 45 without using a special tool such as a pipette.

[0083] The first reagent 101 may contain, for example, a primer having a sequence complementary to the sequence of a nucleic acid molecule derived from the target nucleic acid molecule. The primer may be added as needed as long as it is capable of amplifying the target nucleic acid molecule, and may be, for example, one type or three or more types. The primers may be, for example, a first primer and a second primer. For example, the first primer may be a forward primer that amplifies the target nucleic acid molecule in the sense direction, and the second primer may be a reverse primer that amplifies the nucleic acid molecule in the antisense direction.

[0084] In addition, the first reagent 101 may contain, for example, deoxynucleoside triphosphates (dNTPs), nucleoside triphosphates (NTPs), trehalose, and an RNase inhibitor. However, if the RNase inhibitor is contained in the bottle portion 22, the first reagent 101 does not need to contain the RNase inhibitor.

[0085] As shown in FIG. 7 , the first reagent 101 may be arranged in the first region 451 as a plurality of first attachments. A dried solution containing the above-described reagent may be arranged in the first region 451 as an attachment of the first reagent 101. This allows the first reagent 101 to dissolve in the liquid when the liquid flowing into the flow path 45 comes into contact with the first attachment. Furthermore, when the first reagent 101 is arranged in the flow path 45 independently as a plurality of attachments, the surface area of ​​the first attachment can be increased. This increases the solubility of the first reagent 101 in the liquid. The first reagent 101 may be arranged in the first region 451 as a plurality of films. The number of first attachments may be a plurality, for example, five or more, or ten or more.

[0086] As shown in FIG. 7 , the plurality of first attachments may be arranged in one direction. This allows the first reagent 101 to be dissolved in the liquid sequentially along the flow of the liquid. The plurality of first attachments may be arranged in one direction along the flow path 45. In this embodiment, the first reagent 101 is arranged in one row as the plurality of first attachments, but may be arranged in two or more rows. Furthermore, the plurality of first attachments may be arranged on the bottom 454 of the flow path 45 so as not to come into contact with the inner wall 455 of the flow path 45. This makes it even easier to dissolve the first reagent 101 in the liquid.

[0087] The first attachment may have a dot shape when viewed in a plan view, as shown in Fig. 7. Alternatively, the first attachment may have a linear shape, an elliptical shape, or a polygonal shape such as a square, a hexagon, or a star shape when viewed in a plan view.

[0088] The second reagent 102 may be, for example, an enzyme such as AMV-RT (Avian Myeloblastosis Virus), RNase H (Ribonuclease H), or T7 RNA polymerase. In addition, the second reagent 102 may contain, for example, trehalose and a surfactant.

[0089] As shown in FIG. 7 , the second reagent 102 may be disposed in the second region 452 as a plurality of second attachments. A dried product of the reagent-containing solution may be disposed in the second region 452 as an attachment of the second reagent 102. This allows the second reagent 102 to dissolve in the liquid when the liquid flowing into the flow path 45 comes into contact with the second attachment. When the second reagent 102 is disposed in the flow path 45 as a plurality of independent attachments, the surface area of ​​the second attachment can be increased. This increases the solubility of the second reagent 102 in the liquid. The second reagent 102 may be disposed in the second region 452 as a plurality of films. The number of second attachments may be any number, for example, five or more, or ten or more.

[0090] As shown in FIG. 7 , the plurality of second attachments may be arranged in one direction. This allows the second reagent 102 to be dissolved in the liquid sequentially along the flow of the liquid. The plurality of second attachments may be arranged in one direction along the flow path 45. In this embodiment, the second reagent 102 is arranged in one row as the plurality of second attachments, but may be arranged in two or more rows. Furthermore, the plurality of second attachments may be arranged on the bottom 454 of the flow path 45 so as not to come into contact with the inner wall 455 of the flow path 45. This makes it even easier to dissolve the second reagent 102 in the liquid.

[0091] The second attachment may have a dot shape when viewed in a plan view, as shown in Fig. 7. Alternatively, the second attachment may have a linear shape, an elliptical shape, or a polygonal shape such as a square, a hexagon, or a star shape when viewed in a plan view.

[0092] 6, the second region 452 may be disposed on the opposite side of the liquid receiving section 41 from the position where the first region 451 is disposed. In other words, the second region 452 may be disposed downstream of the first region 451. Therefore, the liquid flowing through the flow path 45B can be brought into contact with the first reagent 101 and then the second reagent 102.

[0093] 6 , the second region 452 may be disposed closer to the reservoir 46 than the first region 451. In other words, the first region 451, the second region 452, and the reservoir 46 may be disposed in this order along the flow path 45. Therefore, the liquid flowing through the flow path 45B can be brought into contact with the first reagent 101 and the second reagent 102 in this order, and then stored in the reservoir 46.

[0094] When the sample contains a target substance, the liquid containing nucleic acid derived from the target substance can be first brought into contact with the first reagent 101. This allows the nucleic acid derived from the target substance to anneal with the first primer and the second primer. Thereafter, the liquid flowing through the flow path 45B comes into contact with the enzyme contained in the second reagent 102, thereby amplifying the nucleic acid derived from the target substance.

[0095] It is sufficient that a first primer and a second primer are disposed in the first region 451 as the first reagent 101. Therefore, for example, dNTP and NTP may be disposed in the second region 452 as the second reagent 102. Even with such an arrangement, nucleic acid derived from the detection target contained in the liquid flowing through the flow channel 45 can be amplified.

[0096] 7 , the second attachment may be larger than the first attachment. For example, the first attachment and the second attachment may be disposed on the flow path 45 by applying a solution of the first reagent 101 and a solution of the second reagent 102 to the flow path 45. The second reagent 102 may contain a surfactant. Therefore, the solution of the second reagent 102 spreads more easily on the bottom surface of the flow path 45 than the solution of the first reagent 101.

[0097] The total area of ​​the plurality of first deposits can be determined by the amount of solution containing the first reagent 101. The total area of ​​the plurality of second deposits can be determined by the amount of solution containing the second reagent 102. For example, the total area of ​​the plurality of first deposits may be approximately twice the total area of ​​the plurality of second deposits.

[0098] The width of the flow path 45 may be, for example, 220 μm. In this case, if the first deposits are dot-shaped, their diameter may be smaller than the width of the flow path 45. The diameter of the first deposits may be, for example, 50 to 100 μm. The spacing between the first deposits may be 100 to 200 μm. If the second deposits are dot-shaped, their diameter may be smaller than the width of the flow path 45. The diameter of the second deposits may be, for example, 80 to 200 μm. The spacing between the second deposits may be 200 to 250 μm.

[0099] For example, if the volume of the liquid containing the specimen stored in the bottle 22 is approximately 50 nL, approximately 48 nL of the first reagent 101 solution and approximately 24 nL of the second reagent 102 solution are required to amplify the nucleic acid contained in the liquid. In this case, if the application volume per attachment is approximately 0.7 nL, the number of first attachments will be 69 and the number of second attachments will be 34. These volumes of liquid and solution are smaller than when using a PCR (Polymerase Chain Reaction) tube. In other words, by using the flow path substrate 4, the volume of the liquid containing the specimen and the application volume (printing volume) of the first reagent 101 and the second reagent 102 applied to the flow path 45 can be reduced.

[0100] The amounts of the first reagent 101 and the second reagent 102 may vary depending on the detection target. Therefore, the amount of the solution of the first reagent 101, the number of first attachments, the amount of the solution of the second reagent 102, and the number of second attachments may be changed depending on the detection target. Furthermore, the size of the first attachments, the size of the second attachments, the spacing between the first attachments, and the spacing between the second attachments may be adjusted depending on the length of the flow channel 45 in the extension direction and the width of the flow channel 45.

[0101] As described above, nucleic acids contained in the liquid flowing from the liquid receiving section 41 may first react with the first reagent 101 disposed in the first region 451 and then react with the second reagent 102 disposed in the second region 452. This is because the first reagent 101 anneals to the nucleic acids contained in the liquid in order to amplify the nucleic acids contained in the liquid with the second reagent 102. Furthermore, as described above, if the cartridge 2 is once heated to a high temperature and then cooled to a temperature that promotes nucleic acid amplification, the temperature may be lowered before the nucleic acids that have reacted with the first reagent 101 react with the second reagent 102. The distance between the first region 451 and the second region 452 may be determined taking these points into consideration. Consider a case in which the heating section 31 heats the bottle section 22 to a temperature of 80°C to 95°C and the main body section 21 to a temperature of 37°C to 41°C. In this case, the second region 452 may be positioned so that the liquid that has flowed in from the bottle portion 22 via the liquid receiving portion 41 comes into contact with the second region 452 after the temperature of the liquid has sufficiently dropped.

[0102] The first region 451 and the second region 452 may be disposed in a linear portion of the flow path 45. In this case, the first reagent 101 and the second reagent 102 can be easily disposed in the flow path 45. When it is not possible to dispose the reagents in one linear portion of the flow path 45, like the first region 451 shown in FIG. 7 , the reagents may be disposed in two or more linear portions.

[0103] A labeling substance that specifically binds to the amplified nucleic acid may further be disposed in the flow path 45. If the labeling substance is a fluorescent substance, the labeling substance emits fluorescence when irradiated with excitation light. Therefore, the detection device 3 can measure the intensity of fluorescence emitted by the nucleic acid derived from the detection target by irradiating the storage section 46 with excitation light.

[0104] The labeling substance may be disposed downstream of the first region 451. For example, the labeling substance may be disposed in the second region 452 as the second reagent 102. This allows the labeling substance to efficiently bind to nucleic acid amplified by the enzyme contained in the second reagent 102. The labeling substance may be a molecular beacon.

[0105] The liquid flowing through one of the channels 45A to 45D may function as a negative control. Figure 6 shows an example in which the channel 45A functions as a channel for negative control.

[0106] The flow path 45A may not include the first region 451, but may include at least a labeling substance. In this embodiment, as shown in FIG. 6 , the flow path 45A may include a second region 452 in which at least a labeling substance is disposed. This prevents the nucleic acid from reacting with a primer for amplifying the nucleic acid, even if the liquid flowing through the flow path 45A contains the nucleic acid. Therefore, the detection device 3 can detect the intensity of fluorescence emitted by substances other than the nucleic acid derived from the target nucleic acid by irradiating the liquid flowing through the flow path 45A and stored in the storage section 46A with excitation light. Therefore, the detection device 3 can accurately detect the intensity of fluorescence emitted by the nucleic acid derived from the target nucleic acid amplified by reaction with the first reagent 101 and the second reagent 102 in the flow path 45B, using the intensity of the fluorescence as a reference.

[0107] In this way, in the flow path substrate 4, both the first reagent 101 and the second reagent 102 may not be arranged, and a flow path 45 that is not intended for detecting nucleic acids derived from the detection target may be connected to the branch flow path 42. In addition to the flow path 45A, the flow paths 45C and 45D may also be flow paths that are not intended for detecting nucleic acids derived from the detection target.

[0108] Each of the flow channels 45A to 45D may have a bending region 453A to 453D downstream of the flow channel 45. Each of the flow channels 45A to 45D may have a bending region 453A to 453D downstream of the position where the reagent is disposed or the position where the reagent can be disposed. This allows a liquid in which the reagent is dissolved to flow through the bending region 453. As this liquid flows through the bending region 453, the reagent and the liquid can be mixed. Therefore, the nucleic acid derived from the detection target can react with the reagent efficiently.

[0109] It is sufficient that the bent region 453 is formed in the flow channel 45 in which the first reagent 101 and the second reagent 102 are disposed. In this embodiment, it is sufficient that the bent region 453B is formed in at least the flow channel 45B.

[0110] Reservoir 46 may be connected to flow path 45. As shown in Fig. 6, reservoirs 46A to 46D may be connected to flow paths 45A to 45D, respectively. Reservoir 46 may store the liquid that flows from flow path 45. Reservoir 46 may be part of the flow path. In other words, reservoir 46 may store a mixed fluid containing a specimen and a labeling substance that reacts with the target substance contained in the specimen.

[0111] When the flow channel 45 has the first region 451 and the second region 452, the reservoir 46 may be located downstream of the second region 452 and may store a liquid containing amplified nucleic acid. In the present embodiment, the flow channel 45B has the first region 451 and the second region 452. Therefore, the reservoir 46B may be located downstream of the second region 452B and may store a liquid containing amplified nucleic acid.

[0112] The reservoir 46 can store the liquid flowing from the flow path 45. When the liquid contains nucleic acid derived from the detection target, the reservoir 46 can store the liquid containing the amplified nucleic acid, which is connected to the flow path 45 having the first region 451 and the second region 452. Therefore, the detection device 3 can stably image the amplified nucleic acid.

[0113] The width of the storage section 46 may be larger than the width of the flow path 45. This can improve the measurement accuracy of the intensity of fluorescence emitted from the storage section 46. The volume of the storage section 46 may be a volume equivalent to the amount of liquid received by the liquid receiving section 41. The volume of the storage section 46 may be, for example, approximately 50 nL. In this way, the storage section 46 may be formed on the flow path substrate 4 so that the volume of the storage section 46 is relatively small.

[0114] 6, the first distance D1 between two adjacent reservoirs 46 among the reservoirs 46A to 46D may be greater than the second distance D2 between the two flow paths 45 connected to each of the reservoirs 46. This allows the spacing between the two adjacent reservoirs 46 to be relatively wide. Therefore, the detection device 3 can easily obtain the intensity of the fluorescence emitted from each reservoir 46 through image analysis.

[0115] A reagent may be placed in the reservoir 46. A reagent used for amplifying nucleic acid derived from the detection target may be placed in the reservoir 46. For example, either the first reagent 101 or the second reagent 102 may be placed in the reservoir 46. Also, for example, a portion of the first reagent 101 and / or the second reagent 102 may be placed in the reservoir 46.

[0116] When either the first reagent 101 or the second reagent 102 is placed in the reservoir 46, a first reagent 101 or a second reagent 102 different from the reagent placed in the reservoir 46 may be placed in a flow path 45 located upstream from the reservoir 46. For example, when the second reagent 102 is placed in the reservoir 46, the first reagent 101 may be placed in a flow path 45 located upstream from the reservoir 46. That is, the first region 451 may be located in the flow path 45 located upstream from the reservoir 46, and the second region 452 may be located in the reservoir 46. This allows the liquid flowing through the flow path 45B to come into contact with the first reagent 101 and then the second reagent 102.

[0117] (Other Configurations) Additionally, the flow path substrate 4 may have second storage sections 48 downstream of the storage sections 46A to 46D, each connected to the storage sections 46A to 46D. In other words, the second storage section 48 may be disposed at a position farther away from the liquid receiving section 41 than the storage section 46. In this case, when a volume of liquid equal to or greater than the capacity of the storage section 46 flows into the storage section 46, the liquid leaking from the storage section 46 can be stored in the second storage section 48. This reduces the possibility of the liquid flowing back from the storage section 46 into the flow path 45.

[0118] A substance functioning as a positive control may also be placed in the second reservoir 48. For example, a substance functioning as a positive control may be placed only in the second reservoir 48 of the flow path substrate 4 used as a positive control. The substance may be optically compared with the substance in the reservoir 46, and may be, for example, a dye, a luminescent substance, or a fluorescent substance, but is not limited thereto. Furthermore, to verify whether the detection system 1 is functioning properly, a sample intentionally containing the target substance may be passed through the liquid receiving section 41 into the flow path 45 before measuring the optical information of the target sample, and the optical information corresponding to the nucleic acid derived from the target sample may be measured in the second reservoir 48. As shown by reference numeral 1102 in FIG. 2 , the second reservoir 48 can be viewed through the window 211 of the main body 21.

[0119] Furthermore, in at least one of the branched flow paths 45, the flow path substrate 4 may have a sensing substance located at a position farther from the liquid receiving section 41 than the storage section 46, detecting the passage of the liquid flowing through the flow path 45. The flow path 45 may be a space that branches at the branch flow path 42 and then communicates with the second storage section 48 via the storage section 46. For example, the position farther from the liquid receiving section 41 than the storage section 46 refers to the position where the liquid received by the liquid receiving section 41 reaches after flowing through the flow path 45 and passing through the storage section 46. The sensing substance may also be a fluorescent substance that is soluble in the liquid flowing through the flow path 45. In this configuration, when the liquid flowing through the flow path 45 passes the position where the sensing substance is located, the sensing substance dissolves. Therefore, the brightness of the sensing substance decreases. The progress of the liquid can be confirmed by checking the decrease in brightness of the sensing substance. In other words, whether the liquid flowing through the flow path 45 is normal can be determined based on whether the brightness of the sensing substance decreases. The detection substance is not particularly limited as long as it can confirm the passage of the liquid flowing through the flow channel 45 .

[0120] The second reservoir 48 may be connected to at least one of the reservoirs 46 A to 46 D. However, the flow path substrate 4 does not necessarily have to have the second reservoir 48.

[0121] Furthermore, the flow path substrate 4 may have an outlet 49 connected to the flow path 45 downstream of the flow path 45. The outlet 49 may be connected to the storage portion 46 downstream of the storage portion 46. As shown in FIG. 6 , when the flow path substrate 4A has a second storage portion 48, the outlet 49 may be connected to the second storage portion 48 downstream of the second storage portion 48. The outlet 49 is a vent that discharges air or liquid in a space located between the liquid receiving portion 41 and the outlet 49 to the outside of the flow path substrate 4A. The space located between the liquid receiving portion 41 and the outlet 49, i.e., the space located within the flow path substrate 4 including the flow path 45, may be referred to as a flow path.

[0122] (Position of Reagents) In the present embodiment, when the first reagent 101 and the second reagent 102 are placed in the flow path 45, it is sufficient that at least a primer and an enzyme are placed in the flow path 45. Other reagents may be placed at positions other than the flow path 45. For example, dNTP, NTP, RNase inhibitor, trehalose, and surfactant may be placed upstream of the flow path 45. For example, these reagents may be placed at branching positions of the flow paths 45A to 45D. For example, these reagents may be placed in the liquid receiving section 41 and / or the branch flow path 42. Furthermore, for example, a standard substance may be placed downstream of the flow path 45. For example, the standard substance may be placed in the reservoir 46.

[0123] (Number of Flow Channels, etc.) In the present embodiment, the flow channel substrate 4 includes four flow channels 45, but this is not limited thereto. In the present embodiment, the flow channel substrate 4 only needs to include at least one flow channel 45 having a first region 451 and a second region 452. In the present embodiment, when the flow channel substrate 4 includes a plurality of flow channels 45 each having a first region 451 and a second region 452, these flow channels 45 may function as flow channels used to detect the same target substance. That is, the first reagent 101 disposed in the plurality of flow channels 45 may be the same reagent, and the second reagent 102 disposed in the plurality of flow channels 45 may be the same reagent. Furthermore, when the flow channel substrate 4A includes a plurality of flow channels 45 each having a first region 451 and a second region 452, these flow channels 45 may function as flow channels used to detect different target substances. That is, the first reagent 101 disposed in the plurality of flow channels 45 may be different reagents, and the second reagent 102 disposed in the plurality of flow channels 45 may be different reagents.

[0124] The flow path substrate 4 may include at least one flow path 45 for a positive control. That is, the flow path 45 may include a flow path 45 for flowing a liquid intentionally containing a detection target. The flow path 45 for the positive control may have a first region 451 and a second region 452 arranged from the upstream side. A liquid intentionally containing a detection target that reacts with the first reagent 101 and the second reagent may be flowed through the flow path 45 for the positive control. However, the flow path 45 having the first region 451 and the second region 452 and used for flowing a liquid containing a specimen may be substituted for the flow path 45 for the positive control.

[0125] Furthermore, in at least one of the branched flow paths 45 of the flow path substrate 4, a sensing substance that detects the passage of the mixed fluid flowing through the flow path 45 may be located at a position farther from the liquid receiving section 41 than the storage section 46. The mixed fluid may contain a sample and a labeling substance that reacts with the target substance contained in the sample. For example, the position farther from the liquid receiving section 41 than the storage section 46 refers to the position where the liquid received by the liquid receiving section 41 reaches after flowing through the flow path 45 and passing through the storage section 46. The sensing substance may also be a fluorescent substance that is soluble in the liquid flowing through the flow path 45. In this configuration, when the liquid flowing through the flow path 45 passes the position where the sensing substance is located, the sensing substance dissolves. Therefore, the brightness of the sensing substance decreases. By confirming the decrease in brightness of the sensing substance, the progress of the liquid can be confirmed. In other words, whether the liquid flowing through the flow path 45 is flowing normally can be determined based on whether the brightness of the sensing substance decreases. The sensing substance is not particularly limited as long as it can confirm the passage of the liquid flowing through the flow path 45.

[0126] For example, the intensity measurement section 351 of the control section 35 may measure the luminance of the fluorescence emitted by the detection substance by analyzing the image captured by the imaging section 34 to obtain the luminance.

[0127] The control unit 35 may include an abnormality determination unit that determines whether the brightness of the fluorescence emitted by the sensing substance is lower than a predetermined value. If the brightness of the fluorescence emitted by the sensing substance is equal to or greater than a predetermined value, the abnormality determination unit may determine that there is an abnormality in the progress of the liquid flowing through the abnormal flow path 45. When an abnormality is determined by this determination, the control unit 35 may notify the user that an abnormality has occurred via an output device such as a display unit and / or speaker (not shown), and then terminate the process of determining the presence or absence of the detection target.

[0128] The number of reservoirs 46 may be determined by the number of connected flow paths 45. That is, a plurality of reservoirs 46 may be connected to each of the branched flow paths 45. For example, each of the branched flow paths may contain a reagent for amplifying nucleic acid corresponding to a different detection target and a labeling substance that specifically binds to the amplified nucleic acid. That is, each flow path 45 may correspond to the detection of a different detection target.

[0129] (Number and Position of Internal Standards) The internal standard 47 may be located in a region different from the branch channel 42, the channel 45, and the reservoir 46. That is, the internal standard 47 used for comparison with the labeled substance may be located in a region different from the channel 45 through which a liquid containing a sample and a labeled substance for detecting the target substance contained in the sample flows. This reduces the possibility that the optical information possessed by the internal standard 47 will change due to contact with the liquid, and the reproducibility of detection of the target substance can be improved by comparing the internal standard 47, which has stable optical information, with the labeled substance.

[0130] 6, the internal standard 47 may be disposed in a region not connected to the flow path 45, near the region R1 in which the storage section 46 is disposed. For example, the internal standard 47 may be located closer to the storage section 46 than the liquid receiving section 41. This makes it easier to simultaneously image the storage section 46 and the internal standard 47. Furthermore, the internal standard 47 may be located around or part of the periphery of the region R1 in which the storage section 46 is disposed.

[0131] Furthermore, the number of internal standards 47 may be any number other than five, which is the number shown in the example of FIG. 6 . The number of internal standards 47 can be set arbitrarily. With this configuration, the target substance can be detected based on the detection results of the fluorescence emitted from the multiple internal standards 47. Therefore, the target substance can be detected with high accuracy. Furthermore, the number of internal standards 47 may be less than the number of multiple storage sections 46 provided in the flow path substrate 4. For example, the multiple storage sections 46 may share the internal standard used to determine the presence or absence of the target substance in the multiple storage sections 46.

[0132] Furthermore, the optical information possessed by at least two of the multiple internal standards 47 may be different from each other. The optical information may be, for example, information obtained from a dye, a luminescent substance, or a fluorescent substance. The optical information may be, for example, information such as color or brightness value. The colors of at least two of the multiple internal standards 47 may be different from each other. For example, one internal standard 47 may be green and another internal standard 47 may be red. For example, the brightness value and intensity of one internal standard 47 may be different from the brightness value and intensity of the other internal standard 47. If the internal standard 47 contains a fluorescent substance, the amount of the fluorescent substance contained in at least two of the multiple internal standards 47 may be different from each other. By using internal standards 47 having different optical information from each other and comparing the multiple optical information with the optical information from the labeling substance, detection of the target substance can be performed with higher accuracy.

[0133] For example, the amount of fluorescent substance contained in one internal standard 47 may be any multiple of the amount of fluorescent substance contained in another internal standard 47. When creating a calibration curve using the fluorescence intensities of the internal standards 47, an appropriate number of internal standards 47 may be provided for creating the calibration curve, and the amount of fluorescent substance contained in each internal standard 47 may be set to an amount with a difference appropriate for creating the calibration curve. By using internal standards 47 having different optical information from each other and comparing the optical information from the multiple internal standards 47 with the optical information from the labeling substance, the target substance can be detected with higher accuracy.

[0134] Furthermore, the luminance emitted by at least one of the multiple internal standards 47 may be set to be higher than the luminance emitted by the mixed fluid stored in the storage section 46. The mixed fluid may contain a specimen and a labeling substance that reacts with the target substance contained in the specimen. For example, the luminance emitted by a certain internal standard 47 may be 10 times the luminance expected to be emitted by the liquid stored in the storage section 46. This configuration allows the internal standard 47 to be used as a positive control.

[0135] 6, a plurality of internal standards 47 may be arranged outside a region R1 in which a plurality of the storage sections 46 of the flow path substrate 4 are arranged. This configuration reduces the possibility that the fluorescence of the storage sections 46 and the fluorescence of the internal standards 47 will affect each other, while allowing the fluorescence emitted by each to be appropriately received. For example, the internal standard 47 may be arranged at a position outside the region R1 shown in FIG. 6, where the internal standard 47 is captured together with the storage sections 46 in an image captured by the imaging section 34.

[0136] Furthermore, the plurality of storage sections 46 included in the flow path substrate 4 may be positioned between the plurality of internal standards 47. This configuration makes it easier to compare the fluorescence of the storage section 46 with the fluorescence of the internal standard 47. For example, each of the plurality of storage sections 46 may be positioned between the plurality of internal standards 47 within the region R1 shown in Fig. 6. That is, the storage sections 46 and the internal standards 47 may be arranged so that the storage sections 46 and the internal standards 47 are positioned alternately.

[0137] Furthermore, in this embodiment, the internal standard 47 is disposed on the flow path substrate 4, but this is not limiting. The internal standard 47 may be disposed on the surface of the housing of the cartridge 2 instead of on the flow path substrate 4. In this case, the internal standard 47 may be disposed near the window 211 of the main body 21. For example, the internal standard 47 may be disposed at a position where the internal standard 47 disposed on the surface of the housing of the cartridge 2 and the reservoir 46 of the flow path substrate 4 are both captured in an image captured by the imaging unit 34. Such a cartridge 2 can be expressed as follows. The cartridge 2 includes a flow path substrate 4 having a flow path through which a mixed fluid containing a sample and a labeled substance that reacts with the target substance contained in the sample flows, and a housing of the main body 21 that houses the flow path substrate 4. In other words, the mixed fluid is located inside the flow path. The housing of the main body 21 includes an internal standard 47 to be compared with the mixed fluid.

[0138] Furthermore, the detection system can be expressed as a determination system using the cartridge 2 having an internal standard in the housing of the main body 21 as follows. The detection system 1 includes a flow path substrate 4, a housing that stores the flow path substrate 4 and stores an internal standard 47 for comparison with a mixed fluid containing a sample and a labeled substance that reacts with the target substance contained in the sample, and an excitation light source that irradiates the mixed fluid and the internal standard 47 with excitation light. The detection system 1 also includes a positive determination unit 352 that determines the presence or absence of the target substance using optical information from the mixed fluid and the internal standard 47. For example, the positive determination unit 352 determines the presence or absence of the target substance using the intensity of fluorescence emitted by the mixed fluid and the internal standard 47, where the labeled substance and the internal standard include a fluorescent substance. Furthermore, the determination method performed by the detection system 1 using the cartridge 2 as described above can be expressed as including the following steps: (1) a step of irradiating the mixed fluid in the flow path substrate 4 and the internal standard 47 in the housing that stores the flow path substrate 4 and stores the internal standard 47 for comparison with the mixed fluid with excitation light. The flow path substrate 4 includes a flow path through which a mixed fluid containing a sample and a labeling substance that reacts with the target substance contained in the sample flows. That is, the mixed fluid is located inside the flow path. (2) A step of measuring optical information of the mixed fluid and the internal standard 47. For example, in this step, the intensity of fluorescence emitted by the mixed fluid and the internal standard 47 is measured. (3) A step of determining the presence or absence of the target substance using the optical information of the mixed fluid and the internal standard 47. For example, in this step, the presence or absence of the target substance is determined using the intensity of fluorescence emitted by the mixed fluid and the internal standard 47.

[0139] Fig. 8 is a plan view showing a flow path substrate 40, which is another example of a flow path substrate included in the cartridge 2 of the present disclosure. In the flow path substrate 40, the multiple internal standards 47a may be positioned so as to surround at least one storage section 46. Specifically, as shown in Fig. 8, the internal standards 47a may be arranged at the four corners of a region R1 in which the multiple storage sections 46 are arranged. By arranging the internal standards 47a in this manner and using the multiple internal standards 47a as alignment marks, it becomes easier to identify the position of the storage section 46.

[0140] The internal standard 47a may have the same dye, luminescent material, or fluorescent material as the above-described internal standard 47a. In Fig. 8, the internal standard 47a is depicted as having a circular shape, but the shape of the internal standard 47a is not particularly limited.

[0141] 9 is a diagram illustrating a method for specifying the position of the storage section 46 using the internal standard 47a. As described above, the position determination section 350 may specify the positions of the multiple internal standards 47 in the captured image and specify the position of each storage section 46 in the captured image from the specified positions of the internal standards 47. In other words, the position determination section 350 may function as an identification section that specifies the position of the storage section 46, which is the detection target, in the captured image based on the positions of the multiple internal standards 47a.

[0142] The position determination unit 350 may function as an identification unit and first identify the positions of multiple fluorescent spots 70 in the captured image. As shown in the diagram indicated by reference numeral 81 in Fig. 9 , multiple fluorescent spots 70 including spots of the internal standard 47a may be displayed in the captured image. In the example shown in Fig. 9 , there are four internal standards 47a, and they are located at the four vertices of a square.

[0143] In order to detect the plurality of fluorescent spots 70 in the captured image, the position determining section 350 may perform binarization processing and contour extraction on the captured image.

[0144] Next, the position determination unit 350 may select any two of the fluorescent spots 70 included in the captured image. In the diagrams indicated by the reference numerals 82 and 83, the selected fluorescent spots are shown as fluorescent spots 71, 72, 73, and 74. When a square is formed having a line segment connecting the two selected fluorescent spots 71 and 72 or the two selected fluorescent spots 73 and 74 as one side of the square, the position determination unit 350 may determine whether another fluorescent spot 70 exists at the positions corresponding to the remaining two vertices.

[0145] In the example indicated by the reference numeral 84, since no fluorescent spots 70 exist at the positions corresponding to the remaining two vertices, the position determination unit 350 determines that at least one of the fluorescent spots 71 and 72 is not the internal standard 47a. The position determination unit 350 may reselect a combination of two other fluorescent spots 70 and repeat the above-described determination for the reselected two fluorescent spots 70. This method reduces the amount of calculation compared to checking each combination of four vertices one by one.

[0146] On the other hand, in the diagram indicated by the reference numeral 85, fluorescent spots 75 and 76 are present at positions corresponding to the remaining two vertices as fluorescent spot 70. Therefore, the position determination unit 350 determines that fluorescent spots 73, 74, 75, and 76 are internal standards 47a.

[0147] If multiple sets of fluorescent spots 70 corresponding to the vertices of the rectangle indicated by the four internal standards 47a can be identified, the position determination unit 350 may identify the set of fluorescent spots 70 corresponding to the vertices of the rectangle having the longest side as the set of internal standards 47a.

[0148] In this way, the position determination unit 350 may identify, as the internal standards 47a, multiple fluorescent spots 70 that exist at positions corresponding to the vertices of a polygon (reference shape) having multiple predetermined internal standards 47a as its vertices. The shape of the reference shape may be any polygon with three or more vertices. In other words, it is sufficient to provide three or more multiple internal standards 47a. The shape of the reference shape is not particularly limited, but the amount of calculation can be reduced by using a square as the reference shape.

[0149] The position determination unit 350 may use the identified multiple internal standards 47a as alignment marks to identify the position of the storage unit 46 that is the detection target. The positional relationship between the multiple internal standards 47a and the storage unit 46 may be stored in advance in a storage device accessible by the position determination unit 350. The position determination unit 350 can identify the position of each storage unit 46 based on the positional relationship, using the positions of the multiple internal standards 47a as a reference.

[0150] 10 is a flowchart showing an example of a method for manufacturing the flow path substrate 4. This manufacturing method may be performed by a manufacturing apparatus for manufacturing the flow path substrate 4.

[0151] 10 , first, a liquid receiving section 41 and a flow path 45 may be formed on a substrate (S1; forming step). In this embodiment, in addition to the liquid receiving section 41 and the flow path 45, a branch flow path 42, a storage section 46, and a second storage section 48 may also be formed on the substrate.

[0152] The liquid receiving portion 41, the flow path 45, etc. may be formed, for example, as follows. For example, a template having the shapes of the liquid receiving portion 41, the flow path 45, etc. patterned thereon is placed on a substrate, and then a resin is poured into the substrate. After the resin has hardened, the template is removed. After the template is removed, a lid is placed on the hardened resin, thereby forming the liquid receiving portion 41, the flow path 45, etc. on the substrate.

[0153] Next, a reagent may be placed in the flow path 45. In this embodiment, a first reagent 101 may be placed in the flow path 45B (S2; first placing step). The first reagent 101 may be placed in the flow path 45B by applying the first reagent 101 to the flow path 45B. Furthermore, in this embodiment, a second reagent 102 may be placed in the flow paths 45A and 45B (S3; second placing step). The second reagent 102 may be placed in the flow paths 45A and 45B by applying the second reagent 102 to the flow paths 45A and 45B.

[0154] Next, an internal standard 47 may be placed on the substrate (S4). The internal standard 47 may be placed on the substrate by applying a fluorescent substance as the internal standard 47 to the substrate. Next, the fluorescent substance may be applied to the second reservoir 48, thereby placing the fluorescent substance in the second reservoir 48 (S5). The order of the processes from S2 to S5 does not matter. The processes from S2 to S5 may be performed in parallel.

[0155] Fig. 11 is a schematic diagram showing an example of a method for applying the reagent 100 to the flow channel 45 provided in the flow channel substrate 4. Reference numeral 1131 in Fig. 12 is a schematic plan view showing an example of the flow channel 45 to which the reagent 100 has been applied, and reference numeral 1132 in Fig. 12 is a schematic diagram showing another example of a method for applying the reagent 100 to the flow channel 45. The reagent 100 in Figs. 11 and 12 may be the first reagent 101 or the second reagent 102.

[0156] The application of the reagent 100 may be performed using, for example, an inkjet printer. Using an inkjet printer enables fine application of the reagent 100. The inkjet printer may include a head 201 that ejects the reagent 100. As shown by reference numeral 1121 in FIG. 11 , the inkjet printer may eject the reagent 100 from the head 201, thereby adhering the reagent 100 to the flow path 45 as shown by reference numeral 1122 in FIG. 11 . The reagent 100 that has adhered to the flow path 45 may be allowed to dry naturally.

[0157] The inkjet printer may apply the reagent 100 while moving the head 201 along the extension direction of the flow path 45, thereby adhering multiple reagents 100 to the bottom 454 of the flow path 45, as shown by reference numeral 1131 in Figure 12.

[0158] The inkjet printer may control the movement of the head 201 and the ejection of the reagent 100 so that the multiple deposits are disposed independently (separately) on the bottom 454. The inkjet printer may also control the movement of the head 201 and the ejection of the reagent 100 so that the deposits do not come into contact with the inner wall 455 of the flow channel 45.

[0159] As indicated by reference numeral 1131 in Fig. 12 , the inkjet printer may control the movement of the head 201 and the ejection of the reagent 100 so that a plurality of deposits are arranged independently side by side in a direction different from the extension direction of the flow channel 45. The plurality of deposits may be arranged side by side, for example, in the width direction of the flow channel 45. Furthermore, as indicated by reference numeral 1132 in Fig. 12 , new reagent 100 may be applied on top of the dried reagent 100 arranged in the flow channel 45.

[0160] [Summary] The flow path substrate according to aspect 1 of the present invention is configured to include a flow path in which a mixed fluid containing a sample and a labeling substance that reacts with the target substance contained in the sample is located, and a standard section that is located in a region different from the flow path and is used for comparison with the mixed fluid.

[0161] A flow path substrate according to Aspect 2 of the present invention may be configured in accordance with Aspect 1 above, further comprising a reservoir connected to the flow path and configured to store the mixed fluid.

[0162] A flow path substrate according to a third aspect of the present invention may be configured in the first or second aspect above, such that the labeling substance and the standard portion contain a fluorescent substance.

[0163] A flow path substrate according to Aspect 4 of the present invention may be configured such that, in Aspect 3 described above, the fluorescent substance contained in the standard portion is a fluorescent substance that is excited by an excitation wavelength that excites the fluorescent substance contained in the labeling substance.

[0164] The flow path substrate of aspect 5 of the present invention may be configured in the above-mentioned aspect 2 to include a liquid receiving section connected to the flow path for receiving the sample, and the standard section is located closer to the storage section than the liquid receiving section.

[0165] A flow path substrate according to Aspect 6 of the present invention may be configured as in any one of Aspects 1 to 5 above, and may include a plurality of the standard portions.

[0166] A flow path substrate according to Aspect 7 of the present invention may be configured in accordance with Aspect 2 above, wherein the flow path substrate includes a plurality of the storage sections, and the number of the standard sections is smaller than the number of the plurality of storage sections.

[0167] The flow path substrate according to aspect 8 of the present invention may be configured in the above-described aspect 2 to include a plurality of the storage sections, and the plurality of standard sections may be arranged outside the area in which the plurality of storage sections are arranged.

[0168] A flow path substrate according to Aspect 9 of the present invention may be configured as in Aspect 2 above, further comprising a plurality of the storage sections, and the plurality of storage sections may be positioned between a plurality of the standard sections.

[0169] A flow path substrate according to aspect 10 of the present invention may be configured such that, in any one of aspects 7 to 9 above, the flow path is branched and the multiple storage sections are connected to each of the branched flow paths.

[0170] A flow path substrate according to an eleventh aspect of the present invention may be configured in accordance with the sixth or eighth aspect above, wherein the optical information of at least two of the plurality of standard portions is different from each other.

[0171] The flow path substrate of aspect 12 of the present invention may be configured in the above aspect 2 or any one of claims 7 to 10 such that the brightness emitted by at least one of the plurality of standard portions is higher than the brightness emitted by the mixed fluid stored in the storage portion.

[0172] The flow path substrate of aspect 13 of the present invention may be configured in the above-mentioned aspect 6 or 12 to have three or more standard sections, and the three or more standard sections may be positioned so as to surround at least one of the storage sections.

[0173] A flow path substrate according to aspect 14 of the present invention may be configured in the above-described aspect 10, further comprising a liquid receiving section connected to the flow path for receiving a liquid containing the sample, and in at least one of the branched flow paths, a detection substance for detecting the passage of the mixed fluid flowing through the flow path is located at a position farther from the liquid receiving section than the storage section.

[0174] A cartridge according to Aspect 15 of the present invention may be configured in accordance with Aspect 1 above, further comprising the flow path substrate and a housing that houses the flow path substrate.

[0175] A cartridge according to aspect 16 of the present invention may be configured such that, in the above-mentioned aspect 15, the flow path substrate is connected to the flow path and has a storage section for storing the mixed fluid, and the storage section and the standard section are optically exposed.

[0176] A cartridge according to aspect 17 of the present invention comprises a flow path substrate having a flow path inside which a mixed fluid containing a sample and a labeled substance that reacts with the target substance contained in the sample is located, and a housing that stores the flow path substrate, the housing having a standard section for comparison with the mixed fluid.

[0177] A determination system according to aspect 18 of the present invention is configured to include a flow path substrate having a flow path inside which a mixed fluid containing a sample and a labeled substance that reacts with the target substance contained in the sample is located, and a standard section located in an area different from the flow path and for comparison with the mixed fluid, and a first determination section that determines the presence or absence of the target substance using optical information from the mixed fluid and the standard section.

[0178] A determination system according to aspect 19 of the present invention comprises a flow path substrate having a flow path inside which a mixed fluid containing a sample and a labeled substance that reacts with the target substance contained in the sample is located, a housing that stores the flow path substrate and has a standard section that is compared with the mixed fluid, and a first determination section that determines the presence or absence of the target substance using optical information from the mixed fluid and the standard section.

[0179] A determination system according to aspect 20 of the present invention may be configured in the above-mentioned aspects 18 or 19 such that the flow path substrate is provided with a plurality of storage sections connected to the flow paths and for storing the mixed fluid, and the first determination section determines the presence or absence of the detection target for each of the storage sections.

[0180] A determination system according to aspect 21 of the present invention may be configured in any one of aspects 18 to 20 above, to include a plurality of standard parts, wherein the optical information of at least two of the plurality of standard parts is different from each other, and the first determination unit determines the presence or absence of the detection target based on the optical information of the standard parts whose optical information is different from each other.

[0181] A determination system according to aspect 22 of the present invention may be configured in any one of aspects 18 to 21 above, wherein the first determination unit determines the presence or absence of the detection target using an image showing the intensity of fluorescence emitted by the mixed fluid and the standard part, and includes a second determination unit that determines whether the position of the image of the fluorescence emitted by the standard part in the image is within a normal range.

[0182] A determination system according to aspect 23 of the present invention may be configured in any one of aspects 19 to 22 above, wherein the flow path substrate comprises a plurality of the standard sections and at least one storage section connected to the flow path and configured to store the mixed fluid, and further comprises an identification section that identifies the position of at least one storage section in an image showing the intensity of the fluorescence emitted by the mixed fluid and the standard sections based on the positions of the plurality of standard sections.

[0183] A determination system according to aspect 24 of the present invention may be configured in the above-mentioned aspect 23 such that the identification unit identifies the position of the at least one storage section based on the position of a polygon having three or more corners identified by three or more of the standard sections in the image.

[0184] A determination method according to aspect 25 of the present invention is a method comprising the steps of: measuring optical information of the mixed fluid and the standard section in a flow path substrate provided with a flow path in which a mixed fluid containing a sample and a labeled substance that reacts with the target substance contained in the sample is located; and a standard section located in an area different from the flow path and for comparison with the mixed fluid; and determining the presence or absence of the target substance using the optical information of the mixed fluid and the standard section.

[0185] A determination method according to aspect 26 of the present invention is a method including the steps of measuring optical information of a mixed fluid containing a sample and a labeled substance that reacts with the target substance contained in the sample in a flow path substrate having a flow path inside which the mixed fluid is located, and a standard portion in a housing that stores the flow path substrate and has a standard portion for comparison with the mixed fluid, and determining the presence or absence of the target substance using the optical information of the mixed fluid and the standard portion.

[0186] [Additional Notes] The invention according to the present disclosure has been described above based on the drawings and examples. However, the invention according to the present disclosure is not limited to the above-described embodiments. In other words, the invention according to the present disclosure can be modified in various ways within the scope of the present disclosure, and embodiments obtained by appropriately combining the technical means disclosed in different embodiments are also included in the technical scope of the invention according to the present disclosure. In other words, it should be noted that a person skilled in the art could easily make various modifications or corrections based on the present disclosure. It should also be noted that these modifications or corrections are included in the scope of the present disclosure.

[0187] REFERENCE SIGNS LIST 1 Detection system (determination system) 2 Cartridge 33 Light irradiation unit (excitation light source) 4 Flow path substrate 41 Liquid receiving unit 45 Flow path 46 Storage unit 47 Internal standard (standard unit) 352 Positive determination unit (first determination unit) 350 Position determination unit (second determination unit, identification unit)

Claims

1. A flow channel containing a sample and a mixed fluid containing a labeling substance that reacts with the target to be detected in the sample, A flow channel substrate comprising a standard section located in a region different from the flow channel and compared with the mixed fluid.

2. The flow channel substrate according to claim 1, comprising a storage section connected to the flow channel and for storing the mixed fluid.

3. The channel substrate according to claim 1 or 2, wherein the labeling substance and the standard portion include a fluorescent substance.

4. The flow channel substrate according to claim 3, wherein the fluorescent substance contained in the standard portion is a fluorescent substance that is excited by the excitation wavelength at which the fluorescent substance contained in the labeling substance is excited.

5. It includes a liquid receiving section that receives the aforementioned sample and is connected to the flow path. The flow path substrate according to claim 2, wherein the standard portion is located closer to the storage portion than the liquid receiving portion.

6. A flow channel substrate according to any one of claims 1, 2, or 5, comprising a plurality of the standard portions.

7. The system comprises multiple storage units, The flow channel substrate according to claim 2, wherein the number of the standard sections is less than the number of the plurality of storage sections.

8. The system comprises multiple storage units, The flow channel substrate according to claim 2, wherein the plurality of standard portions are arranged outside the region in which the plurality of storage portions are arranged.

9. The system comprises multiple storage units, The plurality of storage units are located between the plurality of standard units, The flow channel substrate according to claim 2, wherein the storage portion and the standard portion are arranged so that they alternately position each other.

10. The aforementioned flow path is branched, The flow channel substrate according to any one of claims 7 to 9, wherein the plurality of storage sections are connected to each of the branched flow channels.

11. The flow channel substrate according to claim 6, wherein the optical information of at least two of the plurality of standard sections is different from that of the other.

12. The flow channel substrate according to claim 2, or any one of claims 7 to 9, wherein the brightness emitted by at least one of the plurality of standard parts is higher than the brightness emitted by the mixed fluid stored in the storage part.

13. The standard part comprises three or more of the above-mentioned parts, The flow path substrate according to claim 2, wherein the three or more standard portions are positioned to surround at least one of the storage portions.

14. It includes a liquid receiving section connected to the flow path that receives the liquid containing the aforementioned sample. The flow path substrate according to claim 10, wherein in at least one of the branched flow paths, a sensing substance for detecting the passage of the mixed fluid flowing through the flow path is located at a position where the distance from the liquid receiving portion is greater than that of the storage portion.

15. The flow channel substrate according to claim 1, A cartridge comprising a housing for housing the aforementioned flow channel substrate.

16. The flow channel substrate is connected to the flow channel and includes a storage section for storing the mixed fluid, The cartridge according to claim 15, wherein the storage portion and the standard portion are optically exposed.

17. A channel substrate having a channel in which a mixed fluid containing a sample and a labeling substance that reacts with the target to be detected contained in the sample is located, The system comprises a housing for housing the aforementioned flow channel substrate, The housing is a cartridge equipped with a standard section for comparison with the mixed fluid.

18. A flow channel containing a sample and a mixed fluid containing a labeling substance that reacts with the target to be detected in the sample, A flow path substrate comprising a standard section located in a region different from the flow path and for comparison with the mixed fluid, A determination system comprising a first determination unit that determines the presence or absence of the detection target using the optical information of the mixed fluid and the standard unit.

19. A channel substrate having a channel in which a mixed fluid containing a sample and a labeling substance that reacts with the target to be detected contained in the sample is located, A housing for housing the flow channel substrate, the housing comprising a standard section for comparison with the mixed fluid, A determination system comprising a first determination unit that determines the presence or absence of the detection target using the optical information of the mixed fluid and the standard unit.

20. The flow channel substrate is connected to the flow channel and comprises a plurality of storage sections for storing the mixed fluid, The determination system according to claim 18 or 19, wherein the first determination unit determines whether or not the detection target is present for each of the storage units.

21. The standard section comprises multiple such standard sections, The optical information of at least two of the aforementioned multiple standard sections is different from that of the other. The determination system according to claim 18 or 19, wherein the first determination unit determines the presence or absence of the detection target based on the optical information of the standard unit, which has different optical information from each other.

22. The first determination unit determines the presence or absence of the detection target using an image showing the intensity of fluorescence emitted by the mixed fluid and the standard unit. The determination system according to claim 18 or 19, further comprising a second determination unit for determining whether the position of the fluorescence image emitted by the standard unit in the aforementioned image is within the normal range.

23. The aforementioned flow channel substrate is Multiple standard parts, It comprises at least one storage section connected to the aforementioned flow path for storing the mixed fluid, The determination system according to claim 19, further comprising a identifying unit that identifies the position of at least one storage unit in an image showing the intensity of the mixed fluid and the fluorescence emitted by the standard units, based on the positions of the plurality of standard units.

24. The determination system according to claim 23, wherein the specified part determines the position of at least one storage unit based on the position of a polygon having three or more angles, which is specified by three or more standard parts in the image.

25. A flow channel substrate comprising a flow channel containing a sample and a labeling substance that reacts with the target to be detected contained in the sample, a standard section located in a region different from the flow channel and compared with the mixed fluid, A step of measuring the optical information of the mixed fluid and the standard part, A determination method comprising the step of determining the presence or absence of the object to be detected using the optical information of the mixed fluid and the standard part.

26. A mixed fluid containing a sample and a labeling substance that reacts with the target to be detected contained in the sample is located in a channel substrate having a channel inside, and a standard portion in a housing that houses the channel substrate and has a standard portion for comparison with the mixed fluid, A step of measuring the optical information of the mixed fluid and the standard part, A determination method comprising the step of determining the presence or absence of the object to be detected using the optical information of the mixed fluid and the standard part.