JP2019113425A - Inspection cartridge and inspection device for immunochromatography assay - Google Patents

Abstract

To improve the accuracy of various assays including biochemical analysis, and to provide inspection cartridges and inspection devices that can achieve miniaturization, generalization, and low prices.SOLUTION: The inspection device includes: a sample pad 2 which absorbs a specimen sample containing a test substance; a conjugate pad 3 which is connected to the sample pad 2 and to which a first specific binding substance specifically binding to the test substance and a labeling substance for recognizing the binding between the test substance and the first specific binding substance are fixed; and a carrier 4 which is connected to the conjugate pad 3 and in which a second specific binding substance specifically binding to the test substance is fixed to a portion thereof, a plurality of the carriers 4 being present.SELECTED DRAWING: Figure 2

Description

  The present invention relates to a test cartridge and a test apparatus for immunochromatographic assay used in various assays including a biochemical test.

  In recent years, biochemical analysis systems for detecting or quantifying biological substances such as DNA, enzymes, antigens, antibodies, proteins, viruses and cells, and chemical substances in various fields such as medicine, chemistry, food, chemical products, etc. It is used. For example, in the medical field, examinations of various diseases including cancer are generally performed by an examination using a biochemical sample such as blood.

  Conventionally, cancer and other medical examinations have been conducted exclusively at specialized medical institutions such as hospitals, but in developed countries such as Japan, examinations at medical institutions are carried out against the background of the arrival of a super aging society, etc. The burden on people also tends to increase. For this reason, there is a demand for a shift from in-hospital testing that requires specialized equipment and advanced expertise and skills to home-use testing that can be easily used by the general public, and treatment using a combination of in-hospital testing and home-based testing. There is a growing social need for medical and preventative care. On the other hand, in developing countries, due to circumstances such as the lack of a health care system, there are situations where necessary medical services are not sufficiently provided. As a means for solving such problems, development of a point-of-care testing (POCT) medical device that can be easily performed without relying on a specialist such as a clinical laboratory technician is required.

  One of the most common analytical devices and biochemical analysis techniques is called an immunochromatographic assay. The method will be briefly described. First, a fiber material such as paper or synthetic fiber is used as a carrier, and a carrier coated with a certain antibody or reagent is prepared. Then, a sample such as blood or urine is dropped on the carrier. Then, the reagent in the carrier and the protein in the sample are bound, and the reaction causes color development to be observed at a certain carrier location. Biochemical analysis is realized by detecting the presence or absence of the protein in the sample at this color depth. For example, Patent Document 1 describes a test method and a test apparatus using an immunochromatographic assay method.

  In such an analysis method, it is required to be able to detect an extremely small amount of a test substance with high sensitivity. For example, in Patent Document 2, as a sensitization (amplification) method for enhancing sensitivity, a test substance is developed on a carrier (backing sheet), and then a specific binding is carried out on the carrier by sending a washing solution or amplification solution. The substance other than the labeled substance captured by selective binding is washed, and the amplification solution is sent on the carrier to optically, electrically or magnetically amplify the labeled substance attached to the labeled antibody. An immunochromatographic assay that can measure a small amount of a test substance with high sensitivity by increasing the change is described.

JP 2012-73111 A JP, 2009-287952, A

  However, there is a problem that the analysis chip and the inspection apparatus used in the conventional immunochromatographic assay method are largely difficult to carry. In addition to the need for a detector such as a spectrophotometer for detection of a fluorescent reagent labeled with an antibody in the conventional immunoassay by ELISA, there is a problem that the test apparatus is large. Although the inspection device of Patent Document 1 is small in size, its internal structure is complicated and its convenience is poor. In addition, since it is necessary to perform a certain operation in the measurement, there is also a problem that general spread is difficult. In addition, in a portable immunochromatographic assay method that can be easily tested, there is a problem that quantitative and multi-item testing is difficult.

  Moreover, in the immunochromatographic assay method of patent document 2, there exists a problem that an amplification solution can not be sent autonomously.

  In addition, in the measurement using the immunochromatographic assay method, when the solution containing the test substance is dropped onto the carrier (backing sheet), the solution flows along the carrier by capillary action, but if the amount of the test substance in the solution is large, Clogging occurs in the carrier, making it difficult for the antigen and antibody to bind, and the optically variable quantitativeness is lost. This is called the hook phenomenon, which is one of the causes for the difficulty in quantitative analysis in the immunochromatographic assay. In the conventional analysis chip, there is only one flow path through which the solution in the carrier flows, so even if such a phenomenon occurs, it is difficult to reflect it on the analysis result, and there is a problem that the test result is wrong .

  The present invention has been made in view of the circumstances as described above, and is for an immunochromatographic assay that can improve various analytical accuracy including biochemical examination, and can realize downsizing, generalization, and price reduction. An object of the present invention is to provide an inspection cartridge and an inspection apparatus. In addition, since analysis can be performed by a simpler method than the prior art, there is a feature that anyone can perform on the spot.

  In order to solve the problems described above and achieve the object, the present invention provides a sample pad for absorbing an analyte sample containing a test substance, and a sample pad connected to the sample pad and specifically binding to the test substance A conjugate pad on which the specific binding substance of the present invention and a labeling substance for recognizing the binding between the test substance and the first specific binding substance are immobilized, and the conjugate pad is connected to be specific to the test substance And a carrier to which a second specific binding substance bound to is immobilized on a part thereof, and a plurality of carriers are present.

  Further, according to the present invention, there are provided a sample pad for absorbing an analyte sample containing a test substance, a first specific binding substance which is connected to the sample pad and which specifically binds to the test substance, a test substance and a first substance. And a second specific binding substance which is connected to the conjugate pad and is specifically bound to the test substance, wherein the conjugate pad is immobilized with a labeling substance for recognizing the binding to the specific binding substance of And a carrier fixed to a part of the carrier, wherein the carrier is branched into a plurality of branches.

  Further, according to the present invention, there are provided a sample pad for absorbing an analyte sample containing a test substance, a first specific binding substance which is connected to the sample pad and which specifically binds to the test substance, a test substance and a first substance. And a second specific binding substance which is connected to the conjugate pad and is specifically bound to the test substance, wherein the conjugate pad is immobilized with a labeling substance for recognizing the binding to the specific binding substance of And a carrier immobilized on a part thereof, characterized in that there are a plurality of parts to which the second specific binding substance is immobilized.

  ADVANTAGE OF THE INVENTION According to this invention, it is effective in the ability to improve the precision of various assays including a biochemical analysis, and to implement | achieve size reduction, generalization, and price reduction.

It is a top view of the inspection cartridge concerning a 1st embodiment. It is a top view of the internal structure of the test | inspection cartridge of FIG. It is the sectional view on the AA line of the inspection cartridge of FIG. It is a top view of the inspection device concerning a 1st embodiment. It is side surface sectional drawing of the inspection apparatus of FIG. It is a block diagram which shows the electric constitution of the test | inspection apparatus of FIG. It is a flowchart which shows the test | inspection / analysis method using the test | inspection cartridge concerning 1st Embodiment and a test | inspection apparatus. It is a flowchart which shows the modification of the inspection / analysis method using the inspection cartridge concerning 1st Embodiment and an inspection apparatus. It is a figure which shows the modification of the structure (flow path) of the sample pad concerning the 1st embodiment, a conjugate pad, a support | carrier (backing sheet), and an absorption pad. It is a figure which shows the other modification of the structure (flow path) of the sample pad concerning the 1st embodiment, a conjugate pad, a support | carrier (backing sheet), and an absorption pad. It is a figure which shows the other modification of the structure (flow path) of the sample pad concerning the 1st embodiment, a conjugate pad, a support | carrier (backing sheet), and an absorption pad. It is a figure which shows the other modification of the structure (flow path) of the sample pad concerning the 1st embodiment, a conjugate pad, a support | carrier (backing sheet), and an absorption pad. It is a figure which shows the other modification of the structure (flow path) of the sample pad concerning the 1st embodiment, a conjugate pad, a support | carrier (backing sheet), and an absorption pad. It is a figure which shows the other modification of the structure (flow path) of the sample pad concerning the 1st embodiment, a conjugate pad, a support | carrier (backing sheet), and an absorption pad. It is a top view of the inspection cartridge concerning a 2nd embodiment. It is a top view of the internal structure of the test | inspection cartridge of FIG. FIG. 16 is a cross-sectional view of the inspection cartridge of FIG. 15 taken along line AA.

  The best mode of an inspection cartridge and an inspection apparatus according to the present invention will be described in detail with reference to the accompanying drawings. In the drawings shown below, for convenience of explanation, the scale of each member in the drawings may be described differently.

First Embodiment
The first embodiment will be described with reference to the accompanying drawings.

<Inspection cartridge>
First, the inspection cartridge will be described. 1 is a plan view of the inspection cartridge according to the first embodiment, FIG. 2 is a plan view of the internal structure of the inspection cartridge of FIG. 1, and FIG. 3 is a sectional view of the inspection cartridge of FIG. FIG. The test cartridge 1 according to the first embodiment optically, magnetically or electrically measures a test substance contained in a sample sample using an antigen-antibody reaction or the like.

  The test cartridge 1 includes a sample pad 2, a conjugate pad 3, a carrier (backing sheet) 4, an absorption pad 5, an analyte sample inlet 6, an opening 7, a liquid pack 8, a microchannel 9, an integrated circuit 10, an integrated circuit A control wiring 11, an inspection line measurement electric wiring 12, and a liquid pack valve control electric wiring 13 are provided.

  Then, among the components of the test cartridge 1, the portions to which the test substance is transferred by the immunochromatographic assay method are the sample pad 2, the conjugate pad 3, the carrier (backing sheet) 4, and the absorbent pad 5. Therefore, the so-called flow path is formed by the sample pad 2, the conjugate pad 3, the carrier (backing sheet) 4, and the absorbent pad 5. In FIGS. 2 and 3, four carriers (backing sheets) 4 are arranged in parallel, and one end of each carrier (backing sheet) 4 is connected to the sample pad 2 and the conjugate pad 3 and the other The absorption pad 5 is connected to the end with a tape or the like.

  The sample pad 2 absorbs a sample sample containing a test substance. The sample sample refers to a solution or liquid that may contain a test substance (natural product, compound, biochemical substance, mineral, organic substance, inorganic substance, etc.) that can be analyzed using the test cartridge and test device of the present invention. It is.

  The conjugate pad 3 comprises an antibody that specifically binds to a test substance, a first specific binding substance such as an antigen, and the test substance and the first specific binding substance optically, electromagnetically, or electrically. And a labeled antibody consisting of a labeling substance for recognizing the binding of

  Here, the specific binding substance is a substance that selectively binds (captures) the test substance contained in the sample of the sample. For example, when the test substance is an antigen, the specific binding substance is an antibody. In addition, specific binding substances and test substances are not limited to this example, and proteins, peptides, particles, biochemical substances, minerals and the like can be used.

  Also, the labeling substance is a substance that binds to a specific binding substance in order to optically, electrically and magnetically recognize the binding of the test substance contained in the analyte sample when the specific binding substance is bound. It is. For example, there are gold nanoparticles, magnetic particles, latex particles and the like.

  The carrier (backing sheet) 4 transfers a sample in the target direction by using a liquid transfer phenomenon such as a capillary phenomenon to form a part of a flow path. The carrier (backing sheet) 4 has a specific binding substance that binds to a test substance contained in an analyte sample immobilized (encapsulated) in two parts of the carrier, and the chemistry of the substance at the immobilized location Generate a reaction or an antigen-antibody reaction. The width of each of the four carriers (backing sheets) 4 is 0.01 to 1 mm.

  Thus, the advantage of having two or more carriers (backing sheet) 4 is as follows. For example, when the same specific binding substance is immobilized on each carrier (backing sheet) 4, optical changes and the like can be simultaneously measured at a plurality of locations, which makes it possible to enhance the reliability of the test results. . When different specific binding substances are immobilized on each carrier (backing sheet) 4, a plurality of tests can be performed simultaneously.

  The sample containing the test substance is absorbed by the sample pad 2 and then passes through the conjugate pad 3, where the test substance binds to the labeled antibody (first specific binding substance and labeling substance), and further capillary action Flow to each carrier (backing sheet) 4 by the Each carrier (backing sheet) 4 is a test line 14 in which a capture antibody (a second specific binding substance and a third specific binding substance) that binds to a labeled test substance is encapsulated (fixed). And a control line 15.

  A second specific binding substance is immobilized on the test line 14. Then, the labeled substance is selectively bound by selectively binding the second specific binding substance and the test substance bound to the labeled antibody (first specific binding substance and the labeled substance) contained in the sample sample. Assembling, the optical, magnetic or electrical properties of the labeling substance are changed, and the color is developed on the carrier (backing sheet) 4.

  An example of a labeling substance using an optical measurement method is gold nanoparticles. Gold nanoparticles consist of several tens to several hundreds of gold atoms, and the reflectance of bulk gold is higher as the wavelength is longer, and the color close to yellow is the reflected light. . However, in gold nanoparticles, wine red or red light is reflected because it is reflected in a wavelength range of 400 to 600 nm under the influence of surface plasmon resonance phenomenon (a collective vibration of atoms by incident light). Another labeling substance is FITC (fluorescein etc.) as a fluorescent substance. FITC has the property of reflecting light of a wavelength of about 520 nm when exposed to ultraviolet light, and in the present invention, quantitative measurement of a substance to be measured is performed by measuring the intensity of light of that wavelength.

  When the measurement is performed with a magnetic change, a magnetic substance (ferromagnetic substance, paramagnetic substance, etc.) such as iron or ferrite is used as a labeling substance, but the invention is not limited thereto. The magnetic substance has a size of micrometer or nanometer, and aggregation of the magnetic substance causes a magnetic change, which is measured by a sensor.

  As other labeling substances, latex particles, silver nanoparticles, organic substances, biological substances and the like can be used, but not limited thereto.

  As described above, it is also possible to measure an electrical change because gold nanoparticles and magnetic substances aggregate and change in resistance (or conductivity). In addition, since optical, magnetic, and electrical changes increase in proportion to the amount of the test substance, proteins and the like are quantitatively measured by measuring those changes.

  The third specific binding substance is fixed to the control line 15. Then, the third specific binding substance and the labeled antibody (the first specific binding substance and the labeling substance) contained in the sample sample bind, whereby the labeling substances are assembled, and the optical and magnetic properties of the labeling substance are obtained. Target or electrical characteristics change, and is a region where color is developed on the carrier (backing sheet) 4. Unlike the test line 14, the control line 15 is a colored area for checking whether the antigen-antibody reaction is normally occurring.

  In the present embodiment, the first specific binding substance, the second specific binding substance, and the third specific binding substance use different substances. The reason is that, for example, when a specific binding substance such as an antibody binds to a substance to be captured, binding can not be achieved unless the binding site (portion serving as a keyhole) is different. However, for example, when a goat antibody is used as the first specific binding substance, an antibody that binds to another site on the capture target substance is used as the second specific binding substance, and a person or mouse that binds to the goat antibody And the like can be used as the third specific binding substance. Therefore, it is also possible to use the binding of the goat antibody in the sample sample and the mouse antibody that recognizes the goat antibody immobilized on the carrier as an indicator that the sample sample has passed on the carrier.

  The absorbent pad 5 absorbs excess water of a solution (analyte sample) in which the test substance having passed through the carrier (backing sheet) 4 is contained.

  The materials of the sample pad 2, the conjugate pad 3, the carrier (backing sheet) 4, and the absorbent pad 5 can be generally known ones. For example, insoluble nanofibers, cellulose nanofibers, chemically synthesized fibers, pulp, nitrocellulose membranes, acetylcellulose membranes, polysulfone membranes, polyester-made membranes, nylon membranes, glass fibers, non-woven fabrics, cloths, other fiber materials, etc. Although it can be, it is not limited to these.

  The sample sample inlet 6 is an opening for introducing a sample sample containing a test substance into the sample pad 2. When the inspection cartridge 1 is attached to (inserted into) the inspection device 18 described later, the opening 7 makes the inspection line 14 of the carrier (backing sheet) 4 and the control line 15 against the inspection portion of the inspection device. Exposed.

  The liquid pack 8 holds the amplification solution and the washing solution. Here, the amplification solution is a solution that enhances the measurement sensitivity of the test substance by amplifying the optical, magnetic or electrical change of the labeling substance. In addition, when the specific binding substance or the impurity on the carrier (backing sheet) 4 is nonspecifically bound and the quantitative measurement becomes difficult, the washing solution is used to wash away the qualitative measurement. Solution. The liquid pack 8 incorporates a compressed air pack 16 and a liquid pack valve 17.

  The compressed air pack 16 is a container in which compressed air is contained. The liquid pack valve 17 is a heat fusible material disposed at the boundary between the liquid pack 8 and the compressed air pack 16. The liquid pack valve 17 is used to automatically feed the liquid in the solution pack 8.

  The minute flow path 9 is a minute flow path for sending the liquid in the liquid pack 8 to the sample pad 2. The minute channel 9 is a structure in which a shape such as unevenness is formed on glass, rubber, or plastic using a photolithography technique and an optical shaping technique used for manufacturing a semiconductor, and a liquid or the like flows therethrough. In addition, when the liquid is caused to flow through the minute flow channel 9 to which a chemical substance or the like is applied, the liquid flowing to the minute flow channel 9 reacts with the chemical substance.

  This chemical mainly uses hydrophobic materials or hydrophilic materials. As a reason for using a hydrophobic material, various solutions such as a sample such as blood and a chemical solution may remain in the channel due to the influence of the viscosity of the liquid. Therefore, by applying a hydrophobic material in the microchannel, the residue is eliminated as much as possible. As a hydrophobic material, there are a positive resist (eg, Tokyo Ohka Kogyo: PMER P series etc.) and a negative resist (eg, SU-8, TMMR series etc. of Tokyo Ohka Kogyo).

  When a hydrophilic material is used in the flow channel, the solution is water-soluble and is used when the solution easily passes through the microchannel. Hydrophobic materials, hydrophilic materials and chemicals are not limited to these.

  The mechanism for automatically sending the liquid pack 8 to the sample pad 2 is as follows. The fluid pack valve 17 is physically melted by a high voltage electric signal or pulse supplied from the boosting unit 36 described later, whereby the boundary between the fluid pack 8 and the compressed air pack 16 disappears and the compressed air pack 16 is opened. . The compressed air in the open compressed air pack 16 flows into the liquid pack 8, and the pressure causes the amplification liquid and the cleaning liquid in the liquid pack 8 to flow into the sample pad 2 via the microchannel 9.

  The integrated circuit 10 is an IC for performing chip recognition and the like. The integrated circuit control wiring 11 is a wiring that electrically connects the inspection device 18 and the integrated circuit 10 when the inspection cartridge 1 is mounted on the inspection device 18 described later. The inspection line measurement electrical wiring 12 is a wiring that electrically connects the inspection device 18 and the inspection line 14 when the inspection cartridge 1 is mounted on the inspection device 18 described later. The liquid pack valve control electrical wiring 13 is a wiring that electrically connects the inspection device 18 and the liquid pack valve 17 when the inspection cartridge 1 is attached to the inspection device 18 described later.

<Inspection device>
Next, the inspection apparatus will be described. 4 is a plan view of the inspection apparatus according to the first embodiment, FIG. 5 is a side sectional view of the inspection apparatus of FIG. 4, and FIG. 6 is a block diagram showing an electrical configuration of the inspection apparatus of FIG. is there. The inspection device 18 according to the first embodiment automatically performs an inspection when connected to an information terminal such as a computer after the inspection cartridge 1 is mounted.

  The inspection device 18 includes a control unit 19, an information terminal connection unit 20, a wireless communication module 21, an infrared communication module 22, a microphone 23, a fingerprint sensor 24, a recording unit 25, an information processing unit 26, an information display unit 27, a battery unit 28, A light emitting element 29, a lens 30, a filter 31, a light receiving element 32, an optical correction sheet 33, an electrochemical measurement processing unit 34, a magnetic sensor 35, a boosting unit 36, and an electrical connector 37 are provided.

  The control unit 19 electronically controls the inspection device 18. The control unit 19 is configured by an integrated circuit or the like. The information terminal connection unit 20 electrically connects the inspection device 18 to an information terminal (not shown) such as a computer, a mobile phone, or a smart phone, and supplies power from the information terminal to the inspection device 18. The information terminal connection unit 20 is a USB serial bus interface, but may be another serial bus interface such as IEEE 1394. The wireless communication module 21 performs wireless communication with a physically separated electric device by Wi-Fi or the like. The infrared communication module 22 performs infrared communication with a physically separated electric device. The microphone 23 captures an external sound. The fingerprint sensor 24 performs fingerprint recognition and mechanical detection.

  The recording unit 25 records, in advance, data such as an advertisement and a moving image to be displayed on the information display unit 27 and data received or processed by the inspection device 18. A semiconductor memory or the like is used for the recording unit 25. The information processing unit 26 processes the data received by the inspection device 18 or the data stored in the recording unit 25. The information processing unit 26 is configured by an integrated circuit or the like. The information display unit 27 displays the information processed by the information processing unit 26 or the information recorded in the recording unit 25. The information display unit 27 uses an LED, an LCD, an organic EL element, or the like. The battery unit 28 supplies necessary power to each part of the inspection apparatus 1 through the control unit 19.

  The light emitting element 29 emits light toward the inspection line 14 of the carrier (backing sheet) 4 exposed at the opening 7 of the inspection cartridge 1. As the light emitting element 29, a lamp, an LED or the like is used. The lens 30 condenses the light reflected from the labeling substance of the carrier (backing sheet) 4 exposed at the opening 7 of the inspection cartridge 1 and performs optical correction of the light. The filter 31 further performs optical correction on the light passing through the lens 30. The light receiving element 32 receives the light passing through the filter 31 and measures the optical change of the labeling substance. As the light receiving element 32, a CCD, a cadmium selenide sensor, or the like is used. The optical correction sheet 33 changes the light reflectance at the part where color is developed by the antigen-antibody reaction depending on the humidity and the production quality, and therefore the complementary color intensity and the light of the light of the inspection line 14 on the inspection cartridge 1 for optical correction. Correction is performed by comparing the intensities of complementary colors of light of the correction sheet 33.

  The electrochemical measurement processing unit 34 measures the electrical change of the labeling substance of the test cartridge 1. The magnetic sensor 35 measures the magnetic change of the labeling substance of the test cartridge 1. For example, the biochemical reaction causes the antibody attached to the magnetic particle and the antibody on the carrier to bind to the protein to be measured, and concentration on a part of the carrier causes a magnetic change, which is a magnetic sensor Measure at 35. The boosting unit 36 boosts the voltage of the electric power supplied from the information terminal connection unit 20 to create a high voltage electric signal or pulse, and supplies it to the liquid pack valve control electric wiring 13 of the inspection cartridge 1. The electrical connector 37 is electrically connected to the integrated circuit control wiring 11 of the inspection cartridge, the inspection line measurement electrical wiring 12, and the liquid pack valve control electrical wiring 13.

<Inspection and analysis method using an inspection cartridge and an inspection device>
Next, an inspection and analysis method using the inspection cartridge 1 and the inspection device 18 according to the first embodiment will be described. In the test cartridge according to the present embodiment, for example, an immunoassay by enzyme-linked immunosorbent assay (hereinafter, ELISA) is possible. As the ELISA method, generally known direct adsorption method, sandwich method and competition method can be used.

  FIG. 7 is a flowchart showing an inspection / analysis method using the inspection cartridge 1 and the inspection apparatus 18 according to the first embodiment. First, a sample is dropped on the sample pad 2 of the test cartridge 1 (step S10). Then, the analyte sample is transmitted from the sample pad 2 to the conjugate pad 3 by capillarity, and the first specific binding substance to which the test substance inside the analyte sample and the labeled substance immobilized in the conjugate pad 3 are bound And then move to the carrier (backing sheet) 4. Then, when the test substance bound to the labeled antibody (the first specific binding substance and the labeling substance) and the second specific binding substance are bound in the test line 14, the labeling substance is agglutinated and the optical property is Changes.

  Next, the inspection cartridge 1 is inserted into the inspection device 18 (step S20). Further, the inspection device 18 is connected to an information terminal such as a computer (step S30). Then, the inspection device 18 receives an electrical signal from the information terminal, the program of the control unit 19 is activated, and the inspection to start the analysis is automatically performed (step S40).

  During analysis, the information display unit 27 displays the advertisement and the moving image recorded in the recording unit 25 (step S50). When the analysis is completed, the information display unit 27 or a display device such as a liquid crystal display of the information terminal displays the analysis result (step S60).

  Next, the control unit 19 or the information terminal confirms the presence or absence of the setting for performing the insurance service or the like (step S70). If there is no setting for performing an insurance service or the like (No at Step S70), the analysis is ended. If there is a setting for performing insurance services etc. (Yes at step S70), trial calculation of insurance and risk calculation analysis etc. are performed based on the analysis data, and the result is displayed on the information display unit 27 or liquid crystal display of the information terminal , Etc. to be presented (step S80), and then the analysis is ended.

(Modification)
FIG. 8 is a flowchart showing a modification of the inspection and analysis method using the inspection cartridge 1 and the inspection device 18 according to the first embodiment. In the modification, step S35 is performed between step S30 and step S40. After the inspection device 18 is connected to an information terminal such as a computer in step S30, when the fingerprint sensor 24 recognizes a fingerprint or detects pressure in step S35, the sensed signal is used as a trigger to control the control unit 19 in step S40. The program starts up and checks are initiated automatically to start the analysis.

<Inspection and analysis method>
The inspection and analysis method in step S40 will be described in detail below. A few minutes after dropping the sample sample onto the test cartridge 1, the program in the control unit 19 causes the power supplied from the information terminal connection unit 20 to be boosted by the boosting unit 36 with a high voltage electrical signal or pulse, It flows to the pack valve control electrical wiring 13. Then, the liquid pack valve 17 is melted by heat, and the solution (cleaning liquid) in the liquid pack 8 flows into the microchannel 9 by the pressure of the compressed air in the compressed air pack 16. The liquid that has flowed into the micro channel 9 flows into the carrier (backing sheet) 4, and the impurities and nonspecifically bound substances trapped in the inspection line 14 on the carrier (backing sheet) 4 flow together with the liquid And absorbed by the absorbent pad 5. Further, when the liquid pack valve 17 of another liquid pack 8 is thermally melted as described above, the solution (amplification liquid) in the liquid pack 8 flows to the carrier (backing sheet) 4 and is captured by the inspection line 14 Optical, magnetic or electrical changes of the labeled substance are amplified.

  This optical, magnetic or electrical change is measured by the optical measurement device (the light emitting element 29, the lens 30, the filter 31, and the light receiving element 32), the magnetic sensor 24, and the electrochemical measurement processing unit 34, respectively. Measure and quantitatively measure the test substance.

  At this time, since the inspection cartridge 1 according to the present embodiment has the inspection line 14 arranged in four parallel, the optical measurement device (the light emitting element 29, the lens 30, the filter 31, and the light receiving element 32) is moved. Since the measurement can be performed without the need, the inspection device 18 can be miniaturized.

  When the reagent (antibody or specific binding substance) held on the carrier is reacted with the sample in the immunochromatographic assay, the concentration of the reaction portion (detection band) of the reagent is proportional to the amount of protein contained in the sample. The reagent is a substance used for analysis, inspection, test, etc., and it is mixed with a predetermined solution as needed to improve fluidity from the viewpoint of ease of operation in inspection, etc. May be used. In the present embodiment, the reaction substance (detection) is obtained by binding the test substance, the labeling substance and the first specific binding substance to the second specific binding substance held in the test line 14. It becomes a band). The detection band is illuminated by the light emitting element 29, and the intensity of the reflected light of the detection band is measured by the light receiving element 32, thereby measuring the concentration of the reaction portion (detection band).

When measuring the chemical substance in the sample, the reagent contained in the carrier of the immunochromatographic assay uses a substance that reacts with the substance in the sample. For example, when it is desired to measure ammonia in a sample, one prepared by immobilizing Nesler's reagent (mixture of mercury (II) iodide and potassium iodide) on a carrier of an immunochromatographic assay is prepared. When measuring in this system, when the sample is dropped on the carrier of the immunochromatographic assay, ammonia in the sample and the Nessler indicator react, and a brown substance (HgO · Hg (NH ₂ ) I) rises. By measuring the floating substance optically, quantitative measurement of the substance in the sample is performed. In addition, in this reacted portion, electron transfer occurs by the chemical reaction, and it becomes possible to measure the change in electrical characteristics, so that the measurement can also be performed by the electrochemical measurement unit. The reagents contained in the substance to be measured and the carrier of the immunochromatographic assay are not limited to these.

  For example, when optical measurement is performed, optical detection of the detection band is performed. In that case, the optical change of the labeling substance attached to the second specific binding substance immobilized on the carrier inspection line 14 is measured. For example, when the labeling substance is gold nanoparticles, the light detected in the inspection line 14 is also wine red because it is optically wine red. In that case, the light emitted from the light emitting element 29 is not absorbed by the inspection line 14, that is, the intensity of the complementary color is measured. By doing so, different from the measurement state of the light source using the conventional mercury lamp etc., other absorbed light can be cut and detected. Therefore, clearer measurement becomes possible.

  The coloration state of the reagent due to the reaction is measured based on the light intensity signal of the inspection line 14 of the inspection cartridge 1, and a filtering process such as a binarization process is applied to measure the amount of protein more accurately. Further, in order to perform correction processing, light from the light emitting element 29 is reflected by the optical correction sheet 33 using the optical correction sheet 33 mounted inside the inspection device 18 and compared with the data measured by the light receiving element 32. Thus, by measuring the storage state and quality state of the immunochromatographic assay, more accurate measurement can be performed. In an optical system related to measurement, an object is to make the measurement of light highly accurate by using a lens, a polarizing filter, a pinhole, or the like, but the use mode is not limited to this.

  As another method, the optical characteristics of the detection band are electrically measured from the light receiving element 32, and quantitative measurement and calculation are performed using the data. For example, if the light source is a green LED that emits light at a wavelength of 555 nm, the RGB code on the program has the values of R60, G2020, and B0. On the other hand, when the complementary color is calculated, R '202, G' 60, and B '262 are obtained, and the wavelength intensity of this value is measured with respect to the data obtained by the CCD to obtain the carrier (backing sheet) 4 (flow path). Since the amount of capture antibody is proportional to the absorption intensity of light, the detection intensity can be measured. In addition, complementary color calculation in color code may be in the form of CMYK, HSV, XYZ, HSL, LAB or the like, but is not limited thereto.

  Since a plurality of carriers (backing sheets) are arranged in parallel, that is, a plurality of flow path structures are provided, it is characterized in that inspection of multiple items can be performed, while the same kind of each flow path When a test is performed using a substance in which a specific binding substance of the present invention and a labeling substance having different optical properties are bound, a change in the optical properties can be measured and quantitative analysis becomes possible.

  Information analysis based on acquired data can be divided into two main stages. In the first step, measurement amount correction of marker proteins of disease is performed. Use the measured amount of the same marker protein obtained at multiple detection points and the measured amount of the standard protein whose abundance does not change regardless of the presence or absence of the disease, in order to accurately determine the protein even when the obtained blood is small Make corrections. In the second step, the corrected amount data of multiple proteins is collated with the disease risk prediction database originally developed to predict the type of disease risk and the degree thereof, and an appropriate coping method. This analysis is not a method to predict disease risk independently from the abundance of individual marker proteins related to conventional diseases, but comprehensively predict disease risk using multivariate analysis using the abundance of multiple proteins I do. By adding the abundance of multiple proteins, it becomes possible to identify the more detailed type and extent of the disease and to provide consulting for appropriate treatment. The obtained specific result is stored in the database together with the diagnosis result at the medical institution. Thus, the prediction accuracy is improved by repeating the prediction analysis.

  In the conventional immunochromatographic assay method, a carrier (flow channel) of about 5 mm in width is prepared, and a certain amount of reaction solution is dropped thereon, or the reaction solution is dropped onto spots in a small shape. There is. However, in this method, since the reaction solution spreads by capillary action, it is difficult to drop a large amount of reaction solution on one test flow channel, so it is difficult to achieve integration and miniaturization.

  However, since the width of the carrier (flow path) is 0.01 to 1 mm, the blood analysis cartridge according to the present invention has a smaller volume of medium and accumulated in the flow path as compared with the conventional immunochromatographic assay using capillary action. Because the amount of wasted sample is reduced, it is possible to measure with a smaller amount of sample than before.

  In the immunochromatographic assay, the carrier (backing sheet) of the test cartridge is configured as a single rectangular sensor. Therefore, when an antigen-antibody reaction occurs in the carrier (backing sheet), the region where color development occurs on the band extends in one direction, and thus movement or correction of the optical system is necessary. On the other hand, in the inspection cartridge and the inspection apparatus according to the first embodiment, only a part of the carrier (backing sheet) develops a color by the antigen-antibody reaction, so that the inspection can be performed without the need to move the optical system. It becomes possible.

(Modification)
Among the configurations of the test cartridge 1 of the present embodiment, the configurations (flow paths) of the sample pad 2, the conjugate pad 3, the carrier (backing sheet) 4, and the absorbent pad 5 can be made different.

  FIG. 9 is a view showing a modified example of the configuration (flow path) of the sample pad 2, the conjugate pad 3, the carrier (backing sheet) 4, and the absorbent pad 5. In FIG. 9, only one carrier (backing sheet) 4 is connected to the conjugate pad 3 and branched to four along the way. Four absorbent pads 5 are connected to each of the four branched ends.

  FIG. 10 is a view showing another modified example of the configuration (flow path) of the sample pad 2, the conjugate pad 3, the carrier (backing sheet) 4, and the absorbent pad 5. In FIG. 9, only one carrier (backing sheet) 4 is connected to the conjugate pad 3 and branched to four along the way. The carrier (backing sheet) 4 merges into one again at the end of the control line 15, and one absorbent pad 5 is connected to the end thereof.

  FIG. 11 is a view showing another modified example of the configuration (flow path) of the sample pad 2, the conjugate pad 3, the carrier (backing sheet) 4, and the absorbent pad 5. In FIG. 11, as in FIG. 9, only one carrier (backing sheet) 4 is connected to the conjugate pad 3 and it branches into four from the middle, but before the carrier (backing sheet) 4 branches. Is formed longer than in FIG. Four absorbent pads 5 are connected to each of the four branched ends.

  FIG. 12 is a view showing another modified example of the configuration (flow path) of the sample pad 2, the conjugate pad 3, the carrier (backing sheet) 4, and the absorbent pad 5. In FIG. 12, two carriers (backing sheets) 4 are connected to the conjugate pad 3. Then, one carrier (backing sheet) 4 is branched into three along the way. One absorbent pad 5 is connected to the end of one carrier (backing sheet) 4, and three absorbent pads 5 are connected to each end of the three branched carrier (backing sheet) 4.

  FIG. 13 is a view showing another modified example of the configuration (flow path) of the sample pad 2, the conjugate pad 3, the carrier (backing sheet) 4, and the absorbent pad 5. In FIG. 13, four conjugate pads 3 are connected to four sample pads 2, and four carriers (backing sheets) 4 are connected to four conjugate pads 3. And one absorption pad 5 is connected to the end of the four carriers (backing sheets) 4.

  FIG. 14 is a view showing another modified example (flow path) of the configuration of the sample pad 2, the conjugate pad 3, the carrier (backing sheet) 4, and the absorbent pad 5. In FIG. 14, one carrier (backing sheet) 4 is connected to one conjugate pad 3, and thereafter, the carrier (backing sheet) 4 forms a spiral and extends to the center of the spiral. On the carrier (backing sheet) 4, four inspection lines 14 and four control lines 15 are formed in order, and an absorbent pad 5 is connected to one end at the center. The spiral shape can absorb blood and other reaction solutions with a small area and volume, thereby preventing excess solution from leaking out of the chip.

Second Embodiment
The second embodiment is different from the first embodiment in the configuration of the inspection cartridge. A second embodiment will be described with reference to the attached drawings. In the configuration of the inspection cartridge according to the present embodiment, parts different from the first embodiment will be described. The other parts are the same as in the first embodiment, so the above description is referred to and the description here is omitted.

  15 is a plan view of an inspection cartridge according to the second embodiment, FIG. 16 is a plan view of the internal structure of the inspection cartridge of FIG. 15, and FIG. 17 is a sectional view of the inspection cartridge of FIG. FIG. The test cartridge 41 according to the second embodiment optically, magnetically or electrically measures a test substance contained in a sample sample using an antigen-antibody reaction or the like.

  The test cartridge 41 includes a sample pad 2, a conjugate pad 3, a carrier (backing sheet) 4, an absorption pad 5, an analyte sample inlet 6, an opening 7, a liquid pack 8, a flow path 9, an integrated circuit 10, integrated circuit control A wiring 11, an inspection line measuring electric wiring 12, and a liquid pack valve control electric wiring 13 are provided. The carrier (backing sheet) 4 includes an inspection line 14 and a control line 15. The liquid pack 8 incorporates a compressed air pack 16 and a liquid pack valve 17.

  In the inspection cartridge 41, two inspection cartridges 1 according to the first embodiment are arranged side by side symmetrically about the integrated circuit 10. As shown in FIG. Thus, except for the integrated circuit 10, all the components are present twice as much as the test cartridge 1.

  Then, after one end of the inspection cartridge 41 is attached (inserted) to the inspection device 18, the inspection described in the first embodiment is performed. After completion of the inspection, the inspection cartridge 41 is removed from the inspection device 18, and the other end of the inspection cartridge 41 is attached again to the inspection device 18, whereby the inspection described in the first embodiment is performed again.

  According to the inspection cartridge and the inspection device according to the second embodiment, since there are two portions to be inspected on both ends on the inspection cartridge, after one of the inspection portions is measured, it is replaced and the other side is replaced. The examination part can be measured. Therefore, many inspections can be performed with one inspection cartridge.

  In the first and second embodiments, the number of carriers (backing sheets) is four, but can be set arbitrarily as long as the test substance can be measured.

  The test cartridge according to the first and second embodiments is based on an enzyme-linked immunosorbent assay (hereinafter referred to as "ELISA"), and methods such as a direct assay, a sandwich assay, and an adsorption assay are not limited thereto. Moreover, DNA, cDNA (complementary DNA), DNA, RNA, etc. can also be measured by the hybridization method, and a to-be-tested substance is not restricted to these.

1, 41 test cartridge 2 sample pad 3 conjugate pad 4 carrier (backing sheet)
DESCRIPTION OF SYMBOLS 5 Absorbent pad 6 Sample sample introduction port 7 Opening 8 Liquid pack 9 Micro flow path 10 Integrated circuit 11 Wiring for integrated circuit control 12 Electric wiring for test line measurement 13 Electric wiring for liquid pack valve control 14 Test line 15 Control line 16 Compression Air pack 17 Liquid pack valve 18 Inspection device 19 Control unit 20 Information terminal connection unit 21 Wireless communication module 22 Infrared communication module 23 Microphone 24 Fingerprint sensor 25 Recording unit 26 Information processing unit 27 Information display unit 28 Battery unit 29 Light emitting element 30 Lens 31 Filter 32 Light receiving element 33 Optical correction sheet 34 Electrochemical measurement processing unit 35 Magnetic sensor 36 Boosting unit 37 Electrical connector

Claims (11)

A sample pad for absorbing a sample sample containing a test substance;
A first specific binding substance, which is connected to the sample pad and specifically binds to the test substance, and a labeling substance for recognizing the binding between the test substance and the first specific binding substance are immobilized With the conjugate pad being
A carrier to which a second specific binding substance, which is connected to the conjugate pad and which specifically binds to the test substance, is immobilized on a part thereof;
The presence of a plurality of the carriers,
Test cartridge for immunochromatographic assay characterized by Branching at least one of the plurality of said carriers into a plurality,
The test cartridge for immunochromatographic assay according to claim 1, characterized in that   The test cartridge for immunochromatographic assay according to claim 1 or 2, wherein a plurality of the sample pads and the conjugate pads are present. A sample pad for absorbing a sample sample containing a test substance;
A first specific binding substance, which is connected to the sample pad and specifically binds to the test substance, and a labeling substance for recognizing the binding between the test substance and the first specific binding substance are immobilized With the conjugate pad being
A carrier to which a second specific binding substance, which is connected to the conjugate pad and which specifically binds to the test substance, is immobilized on a part thereof;
That the carrier is branched into plural,
Test cartridge for immunochromatographic assay characterized by A sample pad for absorbing a sample sample containing a test substance;
A first specific binding substance, which is connected to the sample pad and specifically binds to the test substance, and a labeling substance for recognizing the binding between the test substance and the first specific binding substance are immobilized With the conjugate pad being
A carrier to which a second specific binding substance, which is connected to the conjugate pad and which specifically binds to the test substance, is immobilized on a part thereof;
A plurality of portions to which the second specific binding substance is immobilized,
Test cartridge for immunochromatographic assay characterized by   6. The immunochromatographic assay test cartridge according to claim 5, wherein the carrier is spiral.   The immunochromatographic assay test cartridge according to any one of claims 1 to 6, wherein the width of the carrier is 0.01 to 1 mm.   The test cartridge for immunochromatographic assay according to claim 2 or 4, wherein the width of the branched portion is 0.01 to 1 mm.   The immunochromatographic assay test cartridge according to any one of claims 1 to 8, further comprising an absorption pad that absorbs the sample sample having passed through the carrier.   A test cartridge for immunochromatography assay, characterized in that there are a plurality of test cartridges for immunochromatography assay according to any one of claims 1 to 9.   An immunochromatographic assay test apparatus to which the immunochromatographic assay test cartridge according to any one of claims 1 to 10 is attached.

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