WO2019022299A2 - Puce intégrée de lecture de gène entièrement automatique - Google Patents

Puce intégrée de lecture de gène entièrement automatique Download PDF

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Publication number
WO2019022299A2
WO2019022299A2 PCT/KR2017/011246 KR2017011246W WO2019022299A2 WO 2019022299 A2 WO2019022299 A2 WO 2019022299A2 KR 2017011246 W KR2017011246 W KR 2017011246W WO 2019022299 A2 WO2019022299 A2 WO 2019022299A2
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WIPO (PCT)
Prior art keywords
unit
solution
chamber
mixed
amplification
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PCT/KR2017/011246
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English (en)
Korean (ko)
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WO2019022299A3 (fr
WO2019022299A9 (fr
Inventor
이태재
신수정
임선영
김지현
신설이
이문근
이경균
배남호
이석재
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한국과학기술원
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Publication of WO2019022299A2 publication Critical patent/WO2019022299A2/fr
Publication of WO2019022299A3 publication Critical patent/WO2019022299A3/fr
Publication of WO2019022299A9 publication Critical patent/WO2019022299A9/fr

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    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L7/00—Heating or cooling apparatus; Heat insulating devices
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N35/00—Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor
    • G01N35/08—Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor using a stream of discrete samples flowing along a tube system, e.g. flow injection analysis

Definitions

  • the present invention relates to an integrated automatic gene discrimination chip, and more particularly, to an integrated automatic gene discrimination chip capable of automatically discriminating a gene of a detection target organism.
  • nano-biotechnology capable of directly confirming and manipulating biomolecule behavior of nano units such as genes, proteins and cells of an object have been actively conducted.
  • Such nano-biotechnology can be used to diagnose a disease or to determine the type of a target object, and thus the recent industrial field is being expanded.
  • a lab-on-a-chip is a type of biochip that allows a small chip to perform research that can be done in a laboratory.
  • the lab-on-a-chip is made of plastic, glass, silicon, etc. to create microchannels of nanometer (nm) or less.
  • nm nanometer
  • a typical lab-on-a-chip mixes a PCR buffer and a gene to amplify the gene.
  • the lab-on-a-chip is provided by laminating thin films, there is a problem that the PCR buffer injected between the thin films and the gene are difficult to be mixed quickly.
  • the conventional gene chip is not fully automatic, and it takes a lot of time to identify the gene, which is not practical.
  • An object of the present invention to solve the above problems is to provide an integrated automatic gene discrimination chip capable of automatically discriminating the gene of the detection target bacteria automatically.
  • an extractor for extracting foreign matter from a mixed solution containing a desorption solution and magnetic particles to concentrate the detection target bacteria and extract a gene in the concentrated detection target bacteria;
  • a mixer provided downstream of the concentrating extractor and mixing the gene and a PCR buffer to form a mixed PCR solution;
  • An amplifier provided downstream of the mixer and performing an amplification reaction by performing PCR on the mixed PCR solution;
  • a quantitative injector provided between the mixer and the amplifier for injecting a predetermined amount of the mixed PCR solution into the amplifier;
  • a mixer provided downstream of the amplifier for mixing a signal material with the amplified solution amplified by the amplifier to form a mixed material;
  • a discriminator provided downstream of the mixer for discriminating a gene contained in the mixed material, wherein the discriminator discriminates the gene by measuring the amount of current of the mixed material, .
  • the concentrating extractor includes: a concentrating unit for concentrating the bacteria to be detected by discharging the foreign matter solution from the primary mixed solution in which the desorption solution and the magnetic particles are mixed; And an extracting unit for extracting a foreign substance from the secondary mixed solution in a state in which the bacteria to be detected are concentrated to re-concentrate the bacteria to be detected and extract genes from the bacteria to be detected, wherein the concentrating unit and the extracting unit And a magnetic force is selectively generated to concentrate and extract the bacteria to be detected combined with the magnetic particles.
  • the enrichment unit includes: a concentrating container unit through which the primary mixed solution passes; A concentrating magnet unit provided adjacent to the concentrating container unit and generating a magnetic force selectively in the concentrating container unit; And a condensing motor unit connected to the condensing magnet unit, wherein the condensing motor unit selectively generates a magnetic force in the condensing container unit by adjusting a distance between the condensing magnet unit and the condensing container unit .
  • the enrichment unit may further include a vibration motor unit connected to the concentrating container unit, and the vibration motor unit may include a vibration motor unit, Thereby generating vibration and removing the bacteria to be detected attached to the concentrating container unit.
  • the concentrator may further include a sample mixing unit provided upstream of the thickening unit, and the sample mixing unit may mix the desalination solution and the magnetic particles.
  • the sample mixing unit includes a sample mixing container unit into which the desorption solution and the magnetic particles are injected; A sample mixing heating unit for heating the sample mixing container unit to a predetermined temperature; And a sample mixing motor unit connected to the sample mixing container unit to apply vibration to the sample mixing container unit.
  • the concentrator may further include a concentrator and a foreign matter discharging unit provided downstream of the extracting unit, and the foreign matter discharging unit may discharge the foreign matter solution having passed through the concentrator and the extracting unit So that it can be provided.
  • the extraction unit may include: an extraction container unit through which the secondary mixed solution passes; An extraction magnet unit provided adjacent to the extraction container unit and generating a magnetic force selectively in the extraction container unit; And an extraction motor unit connected to the extraction magnet unit, wherein the extraction motor unit selectively generates a magnetic force to the extraction container unit by adjusting a distance between the extraction magnet unit and the extraction container unit .
  • the extraction unit may include an extraction temperature control unit connected to the extraction container unit and heating and cooling the extraction container unit into which the extraction solution is injected at a predetermined temperature. And an extraction cooling unit connected to the extraction temperature control unit to cool the extraction temperature control unit.
  • the extraction temperature control unit may be characterized in that the extraction container unit is heated and cooled while the magnetic force acting on the extraction container unit is removed.
  • the concentrating extractor further includes an extracting air injecting unit provided upstream of the extracting unit, and the extracted air injecting unit extracts the extracted gene toward a mixer provided downstream of the extracting unit And air is injected into the extraction unit.
  • the extraction air injection unit injects air into the extraction unit in a state where a magnetic force is applied to the extraction unit.
  • the mixer includes a mixer main chamber portion in which the gene and the PCR buffer are accommodated; And a mixer auxiliary chamber portion connected to the mixer main chamber portion and adapted to receive the gene and the PCR buffer accommodated in the mixer main chamber portion, wherein the gene and the PCR buffer are connected to the mixer auxiliary chamber portion And the mixer main chamber portion is reciprocated while being mixed with the mixer main chamber portion.
  • the mixer main chamber portion includes a mixer main chamber main body in which the gene and the PCR buffer are accommodated; And a mixer main chamber extension body provided on one side of the mixer main chamber main body and having a relatively higher stepped portion compared to the mixer main chamber main body.
  • the mixer main chamber portion includes a mixer injection hole formed on a bottom surface of the mixer main chamber main body and into which the gene is injected, and the PCR buffer is inserted into the mixer main chamber main body Wherein the mixer main chamber extension body is filled with the mixture from the mixer main chamber extension to the front end of the mixer injection hole.
  • the mixer main chamber portion may further include a mixer air filter provided on the upper side of the mixer injection hole, the mixer air filter being provided on the upper side of the main mixer main body, And the PCR buffer is mixed with the gene while discharging the bubbles injected together with the gene through the hole.
  • the mixer main chamber part further includes at least one vortex induction unit provided in the main mixer main body, and the vortex induction unit is configured to vortex when it collides with the gene and the PCR buffer And the like.
  • the mixer transfer chamber unit may further include a mixer transfer chamber unit connected to the mixer auxiliary chamber unit.
  • the mixer transfer chamber unit receives the mixed PCR solution in which the gene and the PCR buffer are mixed, And discharging the solution toward the dosing device.
  • the mixer pump unit may further include a mixer pump unit connected to the mixer transfer chamber unit, wherein the mixer pump unit mixes the gene and the PCR buffer to mixer main chamber unit And suction and discharge are performed.
  • the mixer pump unit sucks the mixed PCR solution accommodated in the mixer auxiliary chamber part and accommodates the mixed PCR solution in the mixer transfer chamber part, and mixes the PCR solution contained in the mixer transfer chamber part And air is injected so as to be discharged toward the injector.
  • the quantitative injector may include: a metering chamber having a plurality of metering chambers accommodating a mixed PCR solution formed by mixing a gene and a PCR buffer; And a quantitative stopper portion provided between the pair of adjacent metering chambers and having a plurality of metering stoppers relatively higher in level than the metering chambers.
  • the metering stopper is provided in the metering chamber located upstream, And a step is formed to allow the mixed PCR solution to pass through after the solution is completely filled.
  • the quantitative injector may further include a remaining amount discharging unit for discharging the mixed PCR solution remaining after filling the measuring chamber unit.
  • the quantitative stopper portion may further include a discharge stopper provided between the metering chamber portion and the remaining amount discharging portion and having a relatively higher stepped portion than the metering stopper .
  • the discharge stopper may be formed with a step so that the mixed PCR solution may be passed through the quantitative chamber part before the mixed PCR solution is discharged to the residual amount discharging part .
  • the quantitative injector may further include a quantitative pump unit provided upstream of the quantification chamber unit, and the quantitative pump unit may be configured to transfer the mixed PCR solution contained in the quantification chamber unit to an amplifier .
  • the quantitative air filter unit may further include a quantitative air filter unit provided on the downstream side of the quantification chamber unit, wherein the quantitative air filter unit removes bubbles contained in the mixed PCR solution transferred toward the amplifier can do.
  • the quantitative air filter unit may discharge all of the air injected from the metering pump unit to stop the transfer of the mixed PCR solution when the mixed PCR solution passes through the metering pump unit.
  • the amplifier includes: an amplification film part in which a plurality of films are laminated; An amplification main chamber part provided in the amplification film part and containing a mixed PCR solution formed by mixing a gene and a PCR buffer; A first amplification valve unit provided on the amplification film unit and upstream of the amplification main chamber unit; And a second amplification valve unit provided on the amplification film unit and downstream of the amplification main chamber unit, wherein the mixed PCR solution amplifies the number of genes by amplifying the polymerase chain reaction in the amplification main chamber unit, And a solution is formed.
  • the amplifying film portion may include a main portion of the amplification main chamber portion, the amplification main chamber portion being provided on the upstream side and the downstream side of the main amplification chamber portion, ; And an amplification main chamber film laminated on the valve film and having the amplification main chamber portion formed at a position corresponding to the valve film, wherein the amplification main chamber film covers the entire upper portion of the valve film can do.
  • the amplification main chamber film comprises: an amplification main chamber main film formed with the amplification main chamber portion; And a pair of wing films provided on both sides of the main amplification chamber main film to cover the first amplification valve part and the second amplification valve part, wherein the wing film has a step difference with respect to the main amplification main chamber film And is formed to be high in height.
  • the amplifier includes: an amplification pump unit provided downstream of the amplification main chamber unit and capable of sucking and injecting air so as to transfer an amplification solution in which a gene is amplified; An amplification auxiliary chamber part provided between the amplification pump part and the amplification main chamber part and receiving the amplification solution; And an amplification mixing valve unit provided between the amplification auxiliary chamber unit and the mixer.
  • the amplification pump unit sucks air in a state in which the amplification / mixing valve unit is closed, transfers the amplification solution contained in the amplification main chamber unit to the amplification auxiliary chamber unit, And the air is injected in a state where the amplification / mixing valve is opened, and the amplification solution contained in the amplification assisting chamber is transferred to the mixer.
  • the mixer further comprises: a mixing chamber chamber in which the amplification solution and the signal material are received; And a mixing auxiliary chamber portion connected to the mixing chamber portion and adapted to receive the amplification solution contained in the mixing chamber portion, wherein the amplification solution and the signal material are mixed with the mixing chamber portion and the mixing auxiliary chamber portion, And mixed while reciprocating to form a mixed material.
  • the mixing chamber chamber unit may include a mixing chamber chamber body in which the amplification solution and the signal substance are contained; And a mixed well chamber extension body provided on one side of the mixed well chamber body and having a relatively higher step than the mixed well chamber body.
  • the mixing chamber chamber includes a mixing injection hole formed on the bottom surface of the mixing chamber chamber body for injecting the amplification solution, and the signal material is introduced into the mixing chamber chamber body, The mixture is injected from the mixing chamber chamber extension body and filled up to the front end of the mixing injection hole.
  • the mixing chamber chamber further includes a mixing air filter provided on the upper side of the mixing inlet chamber, the mixing air filter being disposed above the mixing chamber chamber body, And the amplification solution and the signal material are mixed while discharging the bubbles injected together with the amplification solution to the outside.
  • the mixing chamber chamber further includes at least one vortex induction unit provided in the mixing chamber chamber body, wherein the vortex induction unit is configured to generate vortex while colliding with the amplification solution and the signal substance And the like.
  • the amplification solution and the signal material are mixed while reciprocating between the mixing chamber part and the mixing auxiliary chamber part as the discrimination pump part provided in the discriminating device injects and sucks air .
  • the discriminator includes an electrode unit for measuring an amount of current of the mixed material; A discrimination board unit to which a plurality of the electrode units are attached; And a discriminating film unit attached to an upper portion of the discriminating substrate unit and having a reaction chamber through which the mixed substance passes.
  • the electrode unit may include: a working electrode for measuring a current amount of the mixed material; A counter electrode for exchanging electrons derived from the redox reaction of the signal material contained in the mixed material with the working electrode; And a reference electrode having a reference voltage that is a reference of a voltage to be applied to the working electrode, wherein the working electrode measures an amount of current using electrons exchanged with the counter electrode.
  • the present invention provides a gene discrimination apparatus using an integrated automatic gene discrimination chip.
  • the present invention provides a food poisoning discriminator to which an integrated automatic gene discrimination chip is applied.
  • the present invention provides a fish species discriminator to which an integrated automatic gene discrimination chip is applied.
  • the effect of the present invention according to the above configuration is that the bacteria to be detected can be easily concentrated using the magnetic force, and the bacteria to be detected are concentrated in two stages using the concentrating section and the extracting section, Mixing can be prevented. That is, accurate test results can be obtained.
  • the quantitative chamber portion, the quantitative stopper portion, and the remaining amount discharging portion of the present invention can be filled with the predetermined mixed PCR solution in each of the metering chambers without any separate control. That is, the present invention can control the mixed PCR solution injected into the amplifier to be always constant automatically, thereby improving the accuracy of the experiment, and it is economical to inject the mixed PCR solution quickly.
  • the quantitative air filter unit of the present invention eliminates the bubbles contained in the mixed PCR solution transferred toward the amplifier, thereby causing an error in the experimental results due to the air contained in the mixed PCR solution when the mixed PCR solution is amplified Can be prevented.
  • the quantitative air filter unit of the present invention can prevent the mixed PCR solution stored in the amplifier from being pressurized even when the metering pump unit continuously injects air. That is, the quantitative air filter unit is convenient because the metering pump unit can precisely control the time for injecting air to transport the mixed PCR solution.
  • the quantitative support hole of the present invention can prevent a problem that leakage occurs when the film substrate laminated by a plurality of layers spreads when air is injected into the metering pump unit.
  • the amplifying film portion of the amplifier of the present invention does not leak because each film is stacked so as to cover all the upper surfaces of the films laminated on the lower side.
  • the present invention shortens the preparation time for gene discrimination, and makes it possible to quickly identify the gene.
  • FIG. 1 and FIG. 2 are diagrams illustrating the configuration of an integrated automatic gene discrimination chip according to an embodiment of the present invention.
  • FIG. 3 is a combined perspective view of an integrated automatic gene discrimination chip according to an embodiment of the present invention.
  • FIG. 4 is an exploded perspective view of an integrated automatic gene discrimination chip according to an embodiment of the present invention.
  • FIG. 5 is an illustration of a mixer main chamber portion of a blender of an integrated automatic gene discrimination chip according to an embodiment of the present invention.
  • FIG. 6 is an exemplary diagram illustrating a vortex induction unit of a blender of an integrated automatic gene discrimination chip according to an embodiment of the present invention.
  • FIG. 7 is a diagram illustrating an internal fluid flow of a mixer main chamber part of a blender of an integrated automatic gene discrimination chip according to an embodiment of the present invention.
  • FIG. 8 is an exploded perspective view of an amplifier of an integrated automatic gene discrimination chip according to an embodiment of the present invention.
  • FIG. 9 is an exemplary diagram of a discriminator of an integrated automatic gene discrimination chip according to an embodiment of the present invention.
  • FIG. 10 is a flowchart of a gene discrimination method according to an embodiment of the present invention.
  • Fig. 11 is a flowchart of a step of concentrating the detection target bacteria of the gene discrimination method according to an embodiment of the present invention.
  • FIG. 12 is a flowchart illustrating a step of generating a primary mixed solution in the step of concentrating a detection target bacterium in the gene discrimination method according to an embodiment of the present invention.
  • FIG. 13 is a flowchart illustrating a step of injecting a primary mixed solution of a step of concentration of a detection target bacterium in a method for identifying a gene according to an embodiment of the present invention into a concentrating part.
  • FIG. 14 is a flowchart illustrating a step of concentrating the detection target bacteria by discharging a foreign matter solution in the step of concentrating the detection target bacteria in the gene discrimination method according to an embodiment of the present invention.
  • FIG. 15 is a flowchart illustrating a step of transferring a second mixed solution in the step of concentrating the detection target bacteria in the gene discrimination method according to an embodiment of the present invention to the extraction unit.
  • FIG. 16 is a flowchart showing a step of re-concentrating the detection target bacteria in the step of concentrating the detection target bacteria in the gene discrimination method according to an embodiment of the present invention.
  • 17 is a flowchart of a step of extracting a gene of the gene discrimination method according to an embodiment of the present invention.
  • FIG. 18 is a flowchart illustrating a step of extracting a gene from a detection subject bacterium in the step of extracting a gene of the gene discrimination method according to an embodiment of the present invention.
  • FIG. 19 is a flowchart illustrating a step of transferring a gene to a blender in the step of extracting a gene of the gene discrimination method according to an embodiment of the present invention.
  • FIG. 20 is a flowchart of a step of forming a mixed PCR solution of a gene discrimination method according to an embodiment of the present invention.
  • FIG. 21 is a flowchart of a step of injecting a gene according to an embodiment of the present invention.
  • FIG. 22 is a flowchart of a step of forming a mixed material of the gene discrimination method according to an embodiment of the present invention.
  • FIG. 23 is a flowchart of a step of discriminating genes of a gene discrimination method according to an embodiment of the present invention.
  • an extractor for extracting foreign matter from a mixed solution containing a desorption solution and magnetic particles to concentrate the detection target bacteria and extract a gene in the concentrated detection target bacteria;
  • a mixer provided downstream of the concentrating extractor and mixing the gene and a PCR buffer to form a mixed PCR solution;
  • An amplifier provided downstream of the mixer and performing an amplification reaction by performing PCR on the mixed PCR solution;
  • a quantitative injector provided between the mixer and the amplifier for injecting a predetermined amount of the mixed PCR solution into the amplifier;
  • a mixer provided downstream of the amplifier for mixing a signal material with the amplified solution amplified by the amplifier to form a mixed material;
  • a discriminator provided downstream of the mixer for discriminating a gene contained in the mixed material, wherein the discriminator discriminates the gene by measuring the amount of current of the mixed material, .
  • FIG. 1 and FIG. 2 are diagrams illustrating the configuration of an integrated automatic gene discrimination chip according to an embodiment of the present invention.
  • 3 is an assembled perspective view of an integrated automatic gene discrimination chip according to an embodiment of the present invention
  • FIG. 4 is an exploded perspective view of an integrated automatic gene discrimination chip according to an embodiment of the present invention.
  • the integrated automatic gene discrimination chip 1000 includes a concentrator 100, a mixer 200, a quantifier 300, an amplifier 400, a mixer 500, a discriminator 600 A main body chip 700, and a frame machine 800.
  • the concentrator 100 may be provided to extract a foreign object from a mixed solution in which a desalting solution and magnetic particles are mixed to concentrate the detection target bacteria, and to extract a gene in the concentrated detection target bacteria.
  • the buffer unit 120, the concentration pump unit 130, the concentration unit 140, the foreign matter discharge unit 150, And may include a transfer valve unit 190.
  • the mixing unit 110 is provided upstream of the thickening unit 140 to mix the desorption solution and the magnetic particles and includes a mixing vessel unit 111, a mixing heating unit 112, a mixing motor unit 113, And a mixing valve unit (114).
  • the mixing vessel unit 111 may be provided so that the desorption solution and the magnetic particles are injected, and the desorption solution and the magnetic particles are supported for a predetermined time.
  • the desolvation solution may be a solution which has been desorbed from food, fish, etc., but is not limited thereto.
  • the magnetic particles may be bonded to the detection object bacteria contained in the desorption solution, and may be particles having magnetic properties.
  • the mixing heating unit 112 is connected to the mixing container unit 111 and can heat the mixing container unit 111 to a predetermined temperature. At this time, the mixing heating unit 112 may heat the mixing vessel unit 111 to 32 to 42 degrees so that the desorption solution and the magnetic particles can be actively mixed.
  • the mixing motor unit 113 may be connected to the mixing container unit 111 to apply vibration to the mixing container unit 111. Specifically, the mixing motor unit 113 may mix the desalination solution and the magnetic particles contained in the mixing vessel unit 111 by applying vibration to the mixing vessel unit 111 for 20 to 120 minutes .
  • the mixing vessel unit 111 is heated by the mixing heating unit 112 and the mixing motor unit 113 generates vibration to mix the desorption solution and the magnetic particles, The mixing efficiency of the solution and the magnetic particles can be further improved. And, when the desalination solution and the magnetic particles are mixed, the detection subject bacteria contained in the desorption solution can be combined with the magnetic particles.
  • the mixing valve unit 114 is provided between the mixing vessel unit 111 and the thickening unit 140 and is capable of selectively providing the mixed primary solution to the thickening unit 140 . Specifically, the mixing valve unit 114 is closed during the mixing of the desorption solution and the magnetic particles in the mixing vessel unit 111, and when the desorption solution and the magnetic particles are mixed, The mixed solution may be supplied to the concentrating part 140.
  • the buffer unit 120 may be provided upstream of the thickening unit 140 and the buffer unit 120 may be provided to inject the cleaning solution and the transferring solution into the thickening unit 140 .
  • the buffer unit 120 may inject the cleaning solution into the thickening unit 140 so that the foreign matter solution contained in the primary mixed solution may be discharged through the foreign matter discharging unit 150.
  • the buffer unit 120 can transfer the secondary mixed solution stored in the concentrating unit 140 to the extracting unit 180 by injecting the transferring solution into the concentrating unit 140.
  • the secondary mixed solution refers to a solution in which the foreign matter solution is discharged and concentrated in the primary mixed solution.
  • the concentration pump unit 130 may be disposed downstream of the mixing unit 110 and the buffer unit 120 and may be provided upstream of the concentration unit 140.
  • the concentration pump unit 130 may inject the primary mixed solution, the cleaning solution, and the transfer solution into the concentration unit 140.
  • the concentrating unit 140 concentrates the detection target bacteria by discharging the foreign matter solution from the primary mixed solution in which the desorption solution and the magnetic particles are mixed, selectively generates a magnetic force, And is capable of concentrating the bacteria to be detected.
  • the enrichment unit 140 includes a concentration vessel unit 141, a concentrated magnet unit 142, a concentration motor unit 143, a vibration motor unit 144 and a concentration valve unit 145.
  • the concentrating container unit 141 may be provided to receive and pass the primary mixed solution.
  • the concentrating container unit 141 may be provided in the form of a tube, and the liquid injected to one side may be discharged to the other side.
  • the shape of the concentrating container unit 141 is not limited to the embodiment.
  • the concentrated magnet unit 142 is provided adjacent to the concentrating container unit 141 and may generate magnetic force selectively in the concentrating container unit 141.
  • the concentrated magnet unit 142 may be provided below the concentrating container unit 141 and may be provided to apply or remove the magnetic force of the concentrating container unit 141.
  • the condensing magnet unit 142 may be connected to the condensing motor unit 143 to adjust the distance between the condensing magnet unit 142 and the condensing container unit 141. That is, the concentrated magnet unit 142 may move adjacent to the concentrated container unit 141 by the concentrated motor unit 143 to apply a magnetic force to the concentrated container unit 141. Conversely, the condensing magnet unit 142 may be separated from the condensing container unit 141 by the condensing motor unit 143 to remove the magnetic force applied to the condensing container unit 141.
  • the concentrated magnet unit 142 may have a plurality of magnets in the longitudinal direction of the concentrating container unit 141. At this time, the magnets may have different polarities of adjacent magnets. That is, the condensing magnet unit 142 may be provided such that the polarities of the magnets opposed to the concentrating container unit 141 are alternately repeated in the N and S poles.
  • the concentrated magnet unit 142 may be electromagnetically generated to selectively generate a magnetic force in the concentrating container unit 141.
  • a magnetic force may be generated in the concentrated container unit 141 when a current is supplied to the concentrated magnet unit 142.
  • the concentrated motor unit 143 may not be provided.
  • the concentrated magnet unit 142 provided as an electromagnet as described above, since the magnetic force generated in the concentrating container unit 141 can be controlled by controlling the current flowing in the concentrated magnet unit 142, Easy magnetic control is possible.
  • the concentrated magnet unit 142 provided as described above can control the flow of the detection subject bacteria contained in the primary mixed solution by controlling the magnetic force applied to the concentrated container unit 141.
  • the magnetic particles combined with the detection target bacteria are moved toward the concentrated magnet unit 142 side by the magnetic force, And can be fixed to the inner wall of the container unit 141.
  • the vibrating motor unit 144 is connected to the concentrating container unit 141 and generates vibration in the concentrating container unit 141 when the secondary mixed solution is transferred to the extracting unit 180 And the bacteria to be detected attached to the concentrating container unit (141) are peeled off.
  • the secondary mixed solution can maintain the state of being attached to the concentrated magnet unit 142 side in the concentrated vessel unit 141 even when the concentrated vessel unit 141 has its magnetic force removed. Therefore, when the vibration motor unit 144 transfers the secondary mixed solution in the concentrating container unit 141 to the extracting unit 180, Vibration can be applied to the mixed solution to separate from the inner wall.
  • the vibration motor unit 144 provided as described above can increase the transport efficiency of the secondary mixed solution.
  • the foreign matter discharging unit 150 may be disposed downstream of the thickening unit 140 and the extracting unit 180 and the foreign matter discharging unit 150 may include the thickening unit 140 and the extracting unit 180 ) May be provided to discharge the foreign substance solution.
  • the foreign matter discharge unit 150 includes a foreign matter outlet unit 151, a first discharge valve unit 152, and a second discharge valve unit 153.
  • the foreign matter outlet unit 151 is disposed on the downstream side of the concentrating unit 140 and the extracting unit 180 so that the foreign matter solution can be discharged.
  • the first discharge valve unit 152 is provided between the enrichment unit 140 and the discharge port unit 151. When the foreign matter solution having passed through the enrichment unit 140 is transferred to the discharge port unit 151 , And can be opened. The first discharge valve unit 152 may be closed when the second mixed solution is transferred from the enrichment unit 140 to the extraction unit 180.
  • the second discharge valve unit 153 is provided between the extraction unit 180 and the discharge unit 151 so that when the foreign matter solution having passed through the extraction unit 180 is transferred to the discharge unit 151 , And can be opened.
  • the second discharge valve unit 153 may be closed when the gene extracted from the extractor 180 is transferred to the mixer 200.
  • the extraction chamber unit 160 can inject the extraction solution for extracting the genes in the detection target bacteria into the extraction unit 180 and the extraction chamber unit 161, the extraction chamber pump unit 162, Unit 163 and an extraction chamber pressure regulating unit 164.
  • the extraction chamber unit 161 may be provided to receive the extraction solution and may be provided on the upstream side of the extraction unit 180.
  • the extraction chamber pump unit 162 may be connected to the extraction chamber unit 161 to inject air to supply the extraction solution contained in the extraction chamber unit 161 to the extraction unit 180.
  • the extraction chamber valve unit 163 is provided between the extraction chamber unit 161 and the extraction unit 180 and may be provided to control whether the extraction solution contained in the extraction chamber unit 161 is supplied or not have.
  • the extraction chamber pressure regulating unit 164 is connected to the extraction chamber pump unit 162 and the extraction chamber pressure regulating unit 164 is connected to the extraction chamber unit 161 via an air injection tube (Not shown) is connected, the air between the extraction chamber pump unit 162 and the air injection tube is discharged to the outside, and then, the air is sealed.
  • the extraction chamber pressure regulating unit 164 thus provided is configured such that when the air injection tube is connected to the extraction chamber pump unit 162, air between the extraction chamber pump unit 162 and the air injection tube is supplied to the extraction chamber pump It is possible to prevent a problem of flowing into the extraction chamber unit 161 through the unit 162. That is, only a predetermined amount of air can always be injected into the extraction chamber unit 161.
  • the extraction chamber pressure regulating unit 164 may be opened when the extraction chamber pump unit 162 discharges the extraction solution contained in the extraction chamber unit 161 to regulate the internal pressure.
  • the extraction chamber pressure regulating unit 164 thus provided can improve the safety by allowing the air injection tube to be separated from the extraction chamber pump unit 162 while the internal pressure is adjusted.
  • the extraction air injection unit 170 is provided on the upstream side of the extraction unit 180 and may include an extraction air injection unit 171 and an extraction air injection valve 172.
  • the extraction air injection unit 171 injects air into the extraction unit 180 to discharge the gene extracted from the extraction unit 180 toward the mixer 200 provided downstream of the extraction unit 180 can do. At this time, the extraction air injection unit 171 can inject air into the extraction unit 180 in a state where a magnetic force is applied to the extraction unit 180. In this way, when the extraction unit 180 is applied with a magnetic force, the detection target bacteria and the magnetic particles are attached to the extraction unit 180 and fixed. Therefore, in a state where the extraction unit 180 is applied with a magnetic force, only the genes extracted from the detection subject organism can be transferred to the mixer 200.
  • the extraction air injection unit 171 injects air into the extraction unit 180 and transfers the gene in the extraction unit 180
  • the extraction chamber unit 160 is also connected to the extraction unit 180
  • the extraction solution may be further injected to more smoothly transfer the gene.
  • the extracting unit 180 can re-concentrate the detection target bacteria by discharging the foreign matter solution from the secondary mixed solution in a state in which the detection target bacteria are concentrated by the concentration unit 140, The gene to be detected can be extracted.
  • the extraction unit 180 may selectively generate a magnetic force to fix the detection target bacteria combined with the magnetic particles in the extraction unit 180, thereby re-concentrating the detection target bacteria and extracting the gene to be detected have.
  • the extraction unit 180 may include an extraction container unit 181, an extraction magnet unit 182, an extraction motor unit 183, an extraction temperature control unit 184, and an extraction cooling unit 185 .
  • the extraction container unit 181 accommodates the detection target bacteria combined with the magnetic particles, and may be provided so that the secondary mixed solution passes through the extraction container unit 181.
  • the extraction container unit 181 may be provided in a tube shape, but the shape of the extraction container unit 181 is not limited thereto.
  • the extraction magnet unit 182 is provided adjacent to the extraction container unit 181 and can selectively generate magnetic force in the extraction container unit 181.
  • the extraction magnet unit 182 may be provided below the extraction container unit 181 and may be provided to apply or remove the magnetic force of the extraction container unit 181.
  • the extraction magnet unit 182 may be connected to the extraction motor unit 183 to adjust the distance from the extraction container unit 181. That is, the extraction magnet unit 182 may be moved adjacent to the extraction container unit 181 by the extraction motor unit 183 to apply a magnetic force to the extraction container unit 181. Conversely, the extraction magnet unit 182 may be separated from the extraction container unit 181 by the extraction motor unit 183 to remove the magnetic force applied to the extraction container unit 181.
  • the extraction magnet unit 182 may have a configuration in which a plurality of magnets are provided in the longitudinal direction of the extraction container unit 181. At this time, the magnets may have different polarities of adjacent magnets. That is, the extraction magnet unit 182 may be provided such that the polarities of the magnets opposed to the extraction container unit 181 are alternately repeated in the N and S poles. At the intersection of the N pole and the S pole, a strong magnetic flux is formed. Therefore, since the extraction magnet unit 182 captures and captures magnetic particles intensively at the point where the N pole and the S pole intersect, it is possible to prevent the problem that the detection target bacteria combined with the magnetic particles are lost.
  • the extraction magnet unit 182 may be electromagnetically generated to selectively generate magnetic force in the extraction container unit 181.
  • a magnetic force may be generated in the extraction container unit 181 when a current is supplied to the extraction magnet unit 182. Conversely, if the current can not flow through the extraction magnet unit 182, the magnetic force applied to the extraction container unit 181 can be removed. If the extraction magnet unit 182 is provided with an electromagnet, the extraction motor unit 183 may not be provided.
  • the extraction magnet unit 182 provided as described above can control the flow of the detection subject bacteria contained in the secondary mixed solution by controlling the magnetic force applied to the extraction container unit 181.
  • the extraction magnet unit 182 when the extraction magnet unit 182 generates a magnetic force in the extraction container unit 181, the magnetic particles combined with the detection target bacteria are moved toward the extraction magnet unit 182 by the magnetic force, And can be fixed to the inner wall of the container unit 181.
  • the extraction temperature control unit 184 may be connected to the extraction container unit 181 to heat and cool the extraction container unit 181 into which the extraction solution is injected to a predetermined temperature.
  • the extraction temperature control unit 184 can heat and cool the extraction container unit 181 with the magnetic force acting on the extraction container unit 181 being removed so that the temperature of the secondary mixture solution is uniform .
  • the extraction container unit 181 can be heated at 90 to 100 degrees for 5 to 20 minutes. If the temperature of the extraction container unit 181 is less than 90 degrees, the extraction solution may not actively active. If the temperature of the extraction container unit 181 exceeds 100 degrees, the gene may be destroyed, The function of the solution may be lost. Thus, the extraction container unit 181 can be heated from 90 to 100 degrees for 5 minutes to 20 minutes.
  • the extraction solution contained in the extraction container unit 181 heated in an appropriate range can actively break down the cells of the detection target bacteria to rapidly extract genes.
  • the temperature of the extraction container unit 181 is not limited to that described above, and it may be maintained at room temperature depending on the type of extraction solution.
  • the extraction temperature control unit 184 can cool the heated extraction container unit 181 to room temperature. At this time, the extraction temperature control unit 184 may cool the temperature of the extraction container unit 181 by 20 to 30 degrees.
  • the extraction cooling unit 185 may be connected to the extraction temperature control unit 184 to cool the extraction temperature control unit 184.
  • the extraction cooling unit 185 is adapted to be operated simultaneously when the extraction temperature control unit 184 heats the extraction container unit 181 to prevent the failure of the extraction temperature control unit 184 due to overheating And may be operated when the extraction temperature control unit 184 exceeds a predetermined temperature.
  • the extraction cooling unit 184 can operate even when the extraction temperature control unit 184 cools the extraction vessel unit 181 to room temperature so that cooling can be performed more quickly.
  • the extraction unit 180 may further include a first extraction vessel valve 186 and a second extraction vessel valve 187.
  • the first extraction vessel valve 186 and the second extraction vessel valve 187 may be provided adjacent to both sides of the extraction vessel unit 181, respectively.
  • the first extraction vessel valve 186 and the second extraction vessel valve 187 are connected to the extraction vessel unit 181 when the extraction vessel unit 181 is heated by the extraction temperature control unit 184, It can be controlled to adjust the internal pressure and prevent the contained solution from flowing out.
  • the mixer transfer valve unit 190 is provided downstream of the extraction unit 180 and may be opened when the extracted gene is transferred to the mixer 200.
  • FIG. 5 is a diagram illustrating a mixer main chamber part of a blender of an integrated automatic gene discrimination chip according to an embodiment of the present invention
  • FIG. 6 is a block diagram illustrating a vortex induction unit of a blender of an integrated automatic gene discrimination chip according to an embodiment of the present invention
  • FIG. 7 is an exemplary view illustrating an internal fluid flow of a mixer main chamber portion of a mixer of an integrated automatic gene discrimination chip according to an embodiment of the present invention.
  • the mixer 200 is provided downstream of the concentrating extractor 100, and the gene and the PCR buffer are mixed to form a mixed PCR solution.
  • the mixer 200 includes a mixer main chamber 210, a mixer auxiliary chamber 220, a mixer transfer chamber 230, a mixer pump 240, a mixer pressure regulator 250, And a mixer valve unit 270.
  • the gene and the PCR buffer (polymerase chain reaction buffer) are arranged to be mixed while reciprocating between the mixer auxiliary chamber unit 220 and the mixer main chamber unit 210 . ≪ / RTI >
  • the mixer main chamber portion 210 includes a mixer main chamber body 211, a mixer main chamber extension 212, a mixer injection hole 213, a mixer air filter 214 and a vortex induction unit 215, And the PCR buffer is accommodated and mixed with the gene extracted from the target bacteria.
  • the mixer main chamber main body 211 is provided so that the gene and the PCR buffer can be accommodated therein.
  • the mixer main chamber body 211 may be provided in the main body chip 700 formed by stacking a plurality of films, and a space in which the gene and the PCR buffer can be accommodated may be formed therein.
  • the mixer main chamber extension body 212 extends to one side of the mixer main chamber body 211 and a relatively higher step than the mixer main chamber main body 211 can be formed.
  • the mixer main chamber extension 212 is connected to the mixer sub chamber 220 only when the mixer sub chamber 220 sucks the gene and the PCR buffer introduced into the mixer main chamber body 211. [ A step can be formed so as to be able to be transported.
  • the mixer main chamber extension 212 may prevent the gene and the PCR buffer from moving to the mixer sub-chamber 220 when the mixer sub-chamber 220 does not apply a suction force.
  • the mixer injection hole 213 is formed on the bottom surface of the mixer main chamber body 211 and the extracted gene can be injected. As shown in FIG. 7A, the PCR buffer is injected from the mixer main chamber extension 212 before the gene is introduced into the mixer main chamber body 211, and is injected into the mixer injection hole 213 To the front end of the housing. That is, the mixer injection hole 213 provided in the mixer main chamber body 211 can prevent the PCR buffer pre-filled in the mixer main chamber body 211 from flowing into the mixer injection hole 213 and flowing back May be provided.
  • the mixer air filter 214 is provided on the upper side of the mixer main chamber body 211 and may be provided on the upper side of the mixer injection hole 213.
  • the mixer air filter 214 mixes the gene and the PCR buffer while discharging the bubbles injected together with the gene through the mixer injection hole 213 to the outside.
  • the gene introduced through the mixer injection hole 213 includes bubbles.
  • the mixer air filter 214 can discharge the bubbles introduced together with the gene to the outside. At this time, the bubble may burst and the gene and the PCR buffer may be mixed. That is, the mixer air filter 214 can improve mixing efficiency.
  • the mixer air filter 214 may increase the mixing efficiency as described above while discharging the bubbles generated while the gene and the PCR buffer reciprocate.
  • the vortex induction unit 215 may be provided in the mixer main chamber body 211 and the vortex induction unit 215 may be provided to vortex while colliding with the gene and the PCR buffer when they move . 6, the vortex induction unit 215 may be provided to extend inward from both sides of the mixer main chamber body 211, And is provided. The vortex induction unit 215 may be provided on one side of the mixer main chamber body 211 and on the other side of the mixer main chamber body 211 so as to be offset from each other.
  • the vortex induction unit 215 may be provided to extend vertically inward from both sides of the mixer main chamber body 211, and as shown in FIG. 6 (b) and may be formed to extend diagonally inward from both side surfaces of the mixer main chamber body 211 as shown in Figs.
  • the vortex induction unit 215 may be extended in a curved shape from both sides of the mixer main chamber body 211 toward the inside.
  • the vortex induction unit 215 is not limited to the illustrated form, and may be included in one embodiment as long as the shape and position of vortex can be actively generated when the gene and the PCR buffer move.
  • the vortex induction unit 215 thus provided can improve the mixing efficiency of the gene and the PCR buffer.
  • the mixer main chamber body 211 provided as described above is provided on the main body chip 700 formed by lamination of thin films, the mixing of the gene and the PCR buffer is performed in the same space, .
  • the mixer main chamber part 210 according to the present invention is configured to rapidly mix the gene and the PCR buffer, so that the mixed PCR solution can be rapidly formed.
  • the mixer auxiliary chamber unit 220 is connected to the mixer main chamber unit 210 to receive the gene and the PCR buffer accommodated in the mixer main chamber unit 210.
  • the mixer transfer chamber part 230 is connected to the mixer auxiliary chamber part 220 and the mixer transfer chamber part 230 can be provided to receive a mixed PCR solution in which the gene and the PCR buffer are mixed.
  • the mixer transfer chamber 230 may be connected to the quantitative injector 300 so that the mixed PCR solution can be discharged toward the injector 300.
  • the mixer pump unit 240 is connected to the mixer transfer chamber unit 230 and the mixer pump unit 240 mixes the gene and the PCR buffer with the mixer auxiliary chamber unit 220 and the mixer main chamber unit 210 to reciprocate and mix air.
  • the mixer pump unit 240 sucks the mixed PCR solution contained in the mixer main chamber unit 210 and moves the mixed PCR solution to the mixer auxiliary chamber unit 220, As shown in FIG. 7 (c), air may be injected into the mixed PCR solution contained in the mixer auxiliary chamber 220 and transferred to the mixer main chamber 210. In this way, the mixer pump unit 240 can mix the gene and the PCR buffer between the mixer sub-chamber 220 and the mixer main chamber 210 and mix them.
  • the mixer pump unit 240 may suck the mixed PCR solution contained in the mixer auxiliary chamber unit 220 and store the mixed PCR solution in the mixer transfer chamber unit 230.
  • the mixer pump unit 240 may be configured to inject air to discharge the mixed PCR solution contained in the mixer transfer chamber unit 230 toward the metering injector 300.
  • the mixer pump unit 240 may inject the PCR buffer into the mixer main chamber 210. Specifically, the mixer pump unit 240 may inject a preset amount of PCR buffer into the mixer main chamber 210 before the gene is injected.
  • the mixer pressure regulator 250 is connected to the mixer pump 240 and the mixer pressure regulator 250 includes an air injection tube for injecting air into the mixer pump 240, The air between the mixer pump unit 240 and the air injection tube is discharged to the outside, and then the air is sealed.
  • the air injection tube is connected to the mixer pump unit 240, the air supplied from the mixer pump unit 240 to the air injection tube is supplied to the mixer pump unit 240, Can be prevented. That is, only a predetermined amount of air can always be injected.
  • the mixer pressure regulator 250 may be opened when the mixer pump unit 240 discharges the mixed PCR solution to the quantitative injector 300 to regulate the internal pressure.
  • the mixer pressure regulator 250 may be separated from the mixer pump 240 in a state in which the internal pressure of the mixer pressure regulator 250 is adjusted, thereby improving safety.
  • the mixer support hole 260 may be formed at a position adjacent to the mixer pump 240 and the mixer pressure regulator 250.
  • the mixer support hole 260 may prevent the main body 700 from being deformed when the air flows into the mixer pump 240 and the mixer pressure regulator 250 .
  • the mixer support hole 260 prevents deformation of the main body chip 700, thereby preventing leakage of water.
  • the mixer valve unit 270 is provided between the mixer auxiliary chamber unit 220 and the mixer transfer chamber unit 230 and includes a first mixer valve 271 and a second mixer valve 272.
  • the first mixer valve 271 may control the flow between the mixer auxiliary chamber part 220 and the mixer transfer chamber part 230.
  • the second mixer valve 272 may control the flow between the mixer transfer chamber part 230 and the metering injector 300.
  • the first mixer valve 271 may be configured such that the mixer pump unit 240 sucks the mixed PCR solution contained in the mixer auxiliary chamber unit 220 and transfers the mixed PCR solution to the mixer transfer chamber unit 230 It may be in an open state. At this time, the second mixer valve 272 is closed.
  • the second mixer valve 272 may be opened when the mixer pump unit 240 transfers the mixed PCR solution contained in the mixer transfer chamber unit 230 to the metering injector 300. At this time, the first mixer valve 271 is closed.
  • the quantitative injector 300 is provided between the mixer 200 and the amplifier 400, and can inject the mixed PCR solution in a predetermined amount into the amplifier 400 .
  • the metering injector 300 includes a metering chamber 310, a metering stopper 320, a metering pump 340, a metering support hole 350, a metering valve 360, And a metering air filter unit 370.
  • the quantification chamber 310 has a plurality of quantification chambers for receiving a mixed PCR solution formed by mixing a gene and a PCR buffer.
  • the metering chamber 310 includes a first metering chamber 311, a second metering chamber 312, a third metering chamber 313, a fourth metering chamber 314, and a fifth metering chamber 315, .
  • the first to third quantification chambers 311 to 315 may be provided to have a size capable of accommodating a predetermined amount of the mixed PCR solution.
  • the first quantification chamber 311 is provided on the upstream side to which the mixed PCR solution flows, and the downstream side of the second quantification chamber 311, the third quantification chamber 313,
  • the first metering chamber 314 and the fifth metering chamber 315 may be sequentially arranged.
  • the number of the metering chamber units 310 is not limited to one embodiment but may be plural.
  • the metering stopper 320 is provided between adjacent pair of metering chambers, and has a plurality of metering stoppers having a relatively higher stepped portion compared to the metering chambers.
  • a first metering stopper 321 is provided between the first metering chamber 311 and the second metering chamber 312 and the second metering chamber 312 and the third metering chamber 313 are provided between the first metering chamber 311 and the second metering chamber 312,
  • a second metering stopper 322 is provided.
  • a third metering stopper 323 is provided between the third metering chamber 313 and the fourth metering chamber 314 and between the fourth metering chamber 314 and the fifth metering chamber 315
  • a fourth metering stopper 324 is provided.
  • the quantitative stopper unit 320 may be provided with a stepped portion for allowing the mixed PCR solution to pass therethrough in a state where the mixed PCR solution is filled in the quantitative chamber located upstream.
  • the first quantitative stopper 321 to the fourth quantitative stopper 324 of the quantitative stopper 320 may be formed so as to form steps having the same height. In the path of the mixed PCR solution, .
  • the introduced mixed PCR solution is first directed to the first quantitative stopper 321.
  • the first quantitative stopper 321 has a stepped height, the mixed PCR solution can not pass over the first quantitative stopper 321 and is naturally transferred to the first quantitative chamber 311, And is filled in the first metering chamber 311.
  • the first quantitative chamber 311 is filled with the mixed PCR solution, the water level of the mixed PCR solution rises by the step height of the first quantitative stopper 321, and the first quantitative stopper 321 It passes over.
  • the mixed PCR solution having passed through the first quantitative stopper 321 is directed to the second quantitative stopper 322.
  • the second quantitative stopper 322 also has a stepped height, the mixed PCR solution moves toward the second quantitative chamber 312 without passing over the second quantitative stopper 322, The chamber 312 is filled.
  • the second quantitative chamber 312 is filled with the mixed PCR solution, the water level of the mixed PCR solution rises by the step height of the second quantitative stopper 322, and the second quantitative stopper 322 It passes over.
  • the third PCR chamber 313 and the fourth PCR chamber 314 can be filled with the mixed PCR solution.
  • the quantitative stopper 320 is not provided in a stepped shape and the wall surface of the channel channel is made hydrophobic, the mixed PCR solution can be injected in a predetermined amount into each of the metering chambers in accordance with the difference in fluid resistance in the channel channel. .
  • the remaining amount discharging unit 330 may be provided to discharge the mixed PCR solution remaining after filling the metering chamber unit 310.
  • the metering stopper 320 further includes a discharge stopper 325 provided between the metering chamber 310 and the residual amount discharging unit 330.
  • the discharge stopper 325 is provided between the fifth metering chamber 315 and the remaining amount discharging portion 330, and is relatively higher than the metering stoppers 321, 322, 323, Can be formed.
  • the discharge stopper 325 may be discharged to the remaining amount discharging unit 330 through the mixed PCR solution while the mixed PCR solution is completely filled in the metering chamber 310 So that a stepped portion is formed.
  • the quantitative chamber 310, the quantitative stopper 320, and the residual amount discharger 330 provided as described above may be filled with a predetermined mixed PCR solution in each of the quantitative chambers without any separate control. That is, according to the present invention, the mixed PCR solution injected into the amplifier 400 can be constantly maintained, thereby improving the accuracy of the experiment, and the mixed PCR solution can be injected quickly, which is economical.
  • the metering pump unit 340 is provided on the upstream side of the metering chamber unit 310 and can transfer the mixed PCR solution contained in the metering chamber unit 310 to the amplifier 400. More specifically, the metering pump unit 340 is provided between the metering chamber unit 310 and the metering stopper unit 320. When the metering chamber unit 310 is filled with the mixed PCR solution, So that the mixed PCR solution contained in the quantification chamber 310 can be transferred to the amplifier 400.
  • the quantitative support hole 350 may be formed at a position adjacent to the quantitative pump 340. When the air is injected into the metering pump unit 340, the quantitative support hole 350 may prevent the main body chip 700 from being deformed. Also, the quantitative support hole 350 can prevent the deformation of the main body chip 700, thereby preventing the leakage of water.
  • the metering valve unit 360 is provided on the upstream side of the metering pump unit 340.
  • the metering valve unit 360 may be disposed between the metering pump unit 340 and the metering stopper unit 320.
  • the metering valve unit 360 may be disposed between the metering pump unit 340 and the metering stopper unit 320, When the mixed PCR solution contained in the metering chamber 310 is transferred to the amplifier 400, it may be in a closed state.
  • the quantitative air filter unit 370 is provided on the downstream side of the quantification chamber unit 310 and the quantitative air filter unit 370 is configured to collect bubbles contained in the mixed PCR solution transferred toward the amplifier 400 Can be removed. Specifically, when the amplifier 400 amplifies the gene contained in the mixed PCR solution, if the bubble is included, an error may occur in the experimental result. Therefore, the quantitative air filter unit 370 can remove bubbles contained in the mixed PCR solution transferred toward the amplifier 400, thereby preventing the above-described problems from occurring.
  • the quantitative air filter unit 370 may be configured to discharge all the air injected from the quantitative pump unit 340 to stop the transfer of the mixed PCR solution when the mixed PCR solution passes through the quantitative air filter unit 370 . Specifically, when the quantitative pump unit 340 injects air and the mixed PCR solution contained in the quantification chamber unit 310 is all transferred to the amplifier 400, the quantitative pump unit 340 is injected by the quantitative pump unit 340 The air can be exhausted through the quantitative air filter unit 370.
  • the quantitative air filter unit 370 may prevent the mixed PCR solution contained in the amplifier 400 from being pressurized even when the metering pump unit 340 continuously injects air. Accordingly, the quantitative air filter unit 370 is not required to precisely control the injection time of the air for feeding the mixed PCR solution by the quantitative pump unit 340, which is convenient.
  • FIG. 8 is an exploded perspective view of an amplifier of an integrated automatic gene discrimination chip according to an embodiment of the present invention.
  • the amplifier 400 is provided downstream of the mixer 200, and performs PCR for the mixed PCR solution to form an amplification solution .
  • the amplifier 400 includes an amplification film unit 410, an amplification main chamber unit 420, a first amplification valve unit 430, a second amplification valve unit 440, an amplification pump unit 450, A chamber portion 460, an amplification mixing valve portion 470, and an amplification support hole 480.
  • the amplification main chamber part 420 is provided in the amplification film part 410, and a mixed PCR solution formed by mixing a gene and a PCR buffer may be accommodated.
  • the amplification main chamber 420 may form a channel through which the mixed PCR solution passes, and may be provided such that heating and cooling are continuously performed to generate a polymerase chain reaction.
  • the first amplification valve unit 430 may be provided on the amplification film unit 410 and upstream of the amplification main chamber unit 420.
  • the second amplification valve unit 440 may be provided on the amplification film unit 410 and downstream of the amplification main chamber unit 420.
  • the first amplification valve unit 430 and the second amplification valve unit 440 may be accommodated in the amplification main chamber 420 during the polymerase chain reaction have.
  • the amplification film part 410 is laminated with a plurality of films and the valve film 411 is laminated on the amplification main chamber film 412, the amplifying flow film 413, the flow path dummy film 414, the primary dummy film, A film 417, and a secondary dummy film.
  • the valve film 411 has the amplification main chamber part 420 at a position corresponding to the amplification main chamber part 420 formed in the main body chip 700,
  • the first amplification valve unit 430 and the second amplification valve unit 440 may be formed on the upstream side and the downstream side, respectively.
  • the amplification main chamber film 412 is stacked on the valve film 411 and the amplification main chamber part 420 may be formed at a position corresponding to the valve film 411.
  • the amplification main chamber film 412 may cover the entire upper portion of the valve film.
  • the amplification main chamber film 412 includes an amplification main chamber main film 412a and a wing film 412b.
  • the amplification main chamber part 420 may be formed in the amplification main chamber film 412 in a position and shape corresponding to the amplification main chamber part 420 formed in the valve film 411.
  • the wing films 412b may be provided on both sides of the main amplification main body film 412a and cover the upper portions of the first amplification valve unit 430 and the second amplification valve unit 440 .
  • the blade film 412b may be formed to have a step height higher than the amplification main chamber main film 412a.
  • the amplification channel film 413 is provided to cover the upper part of the amplification main chamber main film 412a and the amplification main chamber part 420 is formed at a position corresponding to the amplification main chamber main film 412a .
  • the amplification channel film 413 may be formed to have a thickness equal to that of the amplification main chamber main film 412a and the wing film 412b.
  • the channel dummy film 414 is stacked so as to cover the entire upper portion of the amplification main chamber film 421.
  • the channel dummy film 414 is provided with the amplification main chamber part 420 may be formed.
  • the primary dummy film is stacked so as to cover the entire upper portion of the channel dummy film 414, and the amplification main chamber part 420, the first amplification valve part 430 And the second amplification valve unit 440 may be formed.
  • the primary dummy film is composed of the first primary dummy film 415 and the second primary dummy film 415, but the primary dummy film may be provided in more than one.
  • the valve dummy film 417 is stacked to cover the entire upper portion of the primary dummy film, and the first amplification valve unit 430 and the second amplification valve unit 440 may be formed.
  • the secondary dummy film is stacked so as to cover the entire upper portion of the valve dummy film 417, and the amplification main chamber part 420, the first amplification valve part 430 And the second amplification valve unit 440 may be formed.
  • the secondary dummy film is composed of the first secondary dummy film 418 and the second secondary dummy film 419, but the secondary dummy film may be provided in more than one.
  • the primary dummy film and the secondary dummy film provided as described above can provide a space for swelling when the temperature of the mixed PCR solution contained in the amplification main chamber part 420 increases. That is, the dummy films provide an inflatable space of the amplification main chamber part 420 to prevent damage and sample leakage.
  • the amplifying film part 410 provided as described above is provided such that the films laminated on the valve film 411 cover the entire upper portion of the valve film 411 so that the liquid does not flow out, .
  • the amplification pump unit 450 is provided downstream of the amplification main chamber 420 and is capable of sucking and injecting air to transfer the amplified amplified solution.
  • the amplification auxiliary chamber part 460 is provided between the amplification pump part 450 and the amplification main chamber part 420 and can accommodate the amplification solution.
  • the amplification mixing valve unit 470 may be provided between the amplification assisting chamber unit 460 and the mixer 500.
  • the amplification support part 480 is provided at a position adjacent to the amplification pump part 450 and injects air into and discharges the amplification pump part 450.
  • the amplification pump unit 450 sucks air in a state in which the amplification / mixing valve unit 470 is closed, and supplies the amplification solution stored in the amplification main chamber unit 420 to the amplification auxiliary chamber unit 460
  • the amplification mixing valve unit 470 is opened and the second amplification valve unit 440 is closed to inject the amplification solution contained in the amplification auxiliary chamber unit 460 into the mixer 500, .
  • the mixer 500 is provided downstream of the amplifier 400 and may mix a signal material with the amplified solution amplified by the amplifier 400 to form a mixed material.
  • the mixer 500 may include a mixing chamber part 510 and a mixing auxiliary chamber part 520.
  • the mixing chamber part 510 includes a mixing chamber main body 511, a mixing chamber extension 512, a mixing injection hole 513, a mixing air filter 514 and a vortex induction unit 515.
  • the mixing chamber chamber body 511 is provided so that the amplification solution and the signal material can be received therein.
  • the mixing chamber chamber body 511 may be provided in the body chip 700 formed by stacking a plurality of films, and a space in which the amplification solution and the signal material can be accommodated may be formed therein.
  • the mixing chamber extension body 512 is provided to extend to one side of the mixing chamber main body 511 and relatively higher steps than the mixing chamber main body 511 can be formed. Specifically, the mixing chamber extension 512 is formed in the mixing auxiliary chamber part 520 only when the amplification solution and the signal material introduced into the mixing chamber main body 511 are sucked by the mixing auxiliary chamber part 520, A stepped portion may be formed so as to be able to be transported. The mixing chamber extension 512 may prevent the amplification solution and the signal material from moving to the mixing auxiliary chamber part 520 when the mixing auxiliary chamber part 520 does not apply a suction force.
  • the mixing hole 513 is formed on the bottom surface of the mixing chamber main body 511 to inject the amplification solution.
  • the signal material is injected from the mixing chamber chamber extension 512 before the amplification solution is introduced into the mixing chamber chamber body 511, And is filled up to the front end of the injection hole 513. That is, the mixing injection hole 513 provided in the mixing chamber main body 511 is positioned at a position where the signal material pre-filled in the mixing chamber chamber body 511 flows into the mixing injection hole 513, As shown in FIG.
  • the mixed air filter 514 is provided on the upper side of the mixing chamber main body 511 and may be provided on the upper part of the mixing injection hole 513.
  • the mixed air filter 514 may mix the amplification solution and the signal material while discharging the bubbles injected together with the amplification solution through the mixing injection hole 513 to the outside.
  • the amplification solution introduced through the mixing hole 513 includes bubbles.
  • the mixed air filter 514 may discharge the bubbles introduced together with the gene. At this time, the amplification solution and the signal material can be mixed with the bubbles. That is, the mixing air filter 514 can improve mixing efficiency.
  • the mixing air filter 514 may discharge the bubbles generated as the amplification solution and the signal material reciprocate to the outside, thereby improving the mixing efficiency as described above.
  • the vortex induction unit is substantially the same as the vortex induction unit 215 of the mixer 200, so that redundant description will be omitted.
  • FIG. 9A is an exploded perspective view of a discriminator of an integrated automatic gene discrimination chip according to an embodiment of the present invention
  • FIG. 9B is a diagram showing a discrimination of an integrated automatic gene discrimination chip according to an embodiment of the present invention Fig.
  • the discriminator 600 is provided downstream of the mixer 500, and can identify a gene contained in the mixed material.
  • the discrimination device 600 can discriminate the gene by measuring the amount of the current of the mixed material and includes an electrode unit 610, a discrimination substrate unit 620, a discrimination film unit 630, a discrimination unit 640 A discrimination pump unit 650, a discrimination pressure regulating unit 660 and a discrimination support hole 670.
  • the electrode unit 610 may be formed of a material such as platinum, gold, carbon, copper, nickel, or silver.
  • the electrode unit 610 may measure the amount of current of the mixed material, and may include a working electrode 611, a counter electrode 612, And includes a reference electrode 613.
  • the working electrode 611 can measure the amount of current of the mixed material.
  • the mixed material is a mixture of the amplification solution and the signal material, and the gene contained in the amplification solution is combined with the signal material. If the number of the genes is smaller than that of the signal substance, the signal substance remains in the mixed substance in a state that the signal substance can not bind to the gene.
  • the working electrode 611 can measure an electrical signal of the signal material that is not thus coupled. Therefore, the amount of current measured by the working electrode 611 can be changed in accordance with the amount of the signal material that is not coupled.
  • the counter electrode 612 can exchange electrons derived from the redox reaction of the signal material included in the mixed material with the working electrode 611. Specifically, the counter electrode 612 can exchange electrons derived from the signal material not associated with the gene of the mixed material through the redox reaction with the working electrode 611. The working electrode 611 can accurately measure the amount of current of the mixed material by measuring the amount of electrons exchanged with the counter electrode 612
  • the reference electrode 613 may be a reference for a voltage to be applied to the working electrode 611.
  • the reference electrode 613 has a reference voltage for each of the mixed materials and the working electrode 611 is adapted to apply a voltage according to a reference voltage of the reference electrode 613, (611) can maintain a relatively constant voltage with respect to the mixed material.
  • the applied voltage applied to the working electrode 611 may be a unique voltage value for causing a redox reaction depending on each signal material included in the mixed material.
  • the applied voltage may be adjusted in consideration of the reference electrode 613 so that the signal material included in the mixed material has a voltage value at which a redox reaction can occur.
  • the display voltage may be a sum of the reference voltage and the applied voltage.
  • the display voltage of the working electrode 611 is a reference voltage of 0.2V And an applied voltage of 0.5V.
  • the electrodes 511, 512 and 513 of the electrode unit 610 include a reaction part 614 contacting the mixed material and a ground part 615 extending from the reaction part 614 and grounded ). ≪ / RTI >
  • the reaction unit 614 may refer to a portion where the working electrode 611, the counter electrode 612, and the reference electrode 613 are in contact with the mixed material.
  • the mixed material may be electrochemically reacted with the electrode unit 610 while passing through the reaction unit 614.
  • the ground unit 615 may refer to a grounded portion of the electrodes 611, 612, and 613 included in the electrode unit 610 and may extend from the reaction unit 614.
  • the grounding unit 615 may be separated from the reaction unit 614 by the discriminating film unit 630 so as not to be in contact with the mixed substance.
  • a plurality of the electrode units 610 may be attached to the upper portion of the identification substrate unit 620.
  • the material of the identification substrate unit 620 may be glass, and a plurality of the electrode units 610 may be attached to the upper portion of the identification substrate unit 620 in the longitudinal direction. At this time, the material of the identification substrate unit 620 is not limited to glass.
  • the discriminating film unit 630 is attached to the upper portion of the discriminating substrate unit 620, and a reaction chamber through which the mixed substance passes may be formed.
  • the determination film unit 630 may be provided to form the reaction chambers at positions corresponding to the plurality of electrode units 610 provided in the longitudinal direction of the identification substrate unit 620.
  • the reaction chamber may be formed in such a shape that mixed material having passed through the mixer 200 flows into one side, reacts with the electrode unit 610, and is discharged to the other side.
  • the width of the discriminating film unit 630 is divided into the reaction part 614 and the grounding part 615 of the electrode unit 610 so that the grounding part 615 and the mixed material are not in contact with each other .
  • the discriminating and discharging unit 640 may be disposed downstream of the discriminating film unit 630 and may receive the mixed substance that has passed through the discriminating film unit 630 and has been subjected to gene discrimination to be discharged to the outside.
  • the discrimination pump unit 650 is connected to the mixing auxiliary chamber unit 520 and the discrimination pump unit 650 amplifies the amplification solution and the signal material from the mixing chamber main body 510 and the mixing auxiliary chamber unit 520) to be mixed and reciprocated.
  • the discrimination pump unit 650 sucks the air in the mixing chamber part 510 to move the amplification solution and the signal material to the mixing auxiliary chamber part 520,
  • the amplification solution and the signal material accommodated in the chamber part 520 can be transferred to the mixing chamber part 510.
  • the discrimination pump unit 650 may mix the amplification solution and the signal material between the mixing auxiliary chamber unit 520 and the mixing chamber chamber unit 510 to form a mixed material.
  • the discrimination pump unit 650 may suck air such that the mixed material contained in the mixing auxiliary chamber unit 520 passes through the discriminating film unit 630 and moves to the discriminating discharging unit 640 .
  • the discrimination pressure regulating unit 660 is connected to the discrimination pump unit 650 and the discrimination pressure regulating unit 660 is connected to the discrimination pump unit 650 through an air suction tube (not shown)
  • the air in the space between the discrimination pump unit 650 and the air suction tube is discharged to the outside and is then sealed.
  • the discrimination pressure regulating unit 660 provides air to the discrimination pump unit 650, It is possible to prevent the problem of flowing into the inside through the through hole.
  • the discrimination pressure regulating unit 660 may be opened when the discrimination pump unit 650 moves and discharges the mixed substances to regulate the internal pressure.
  • the discrimination pressure regulating unit 660 thus provided can separate the air suction tube from the discriminating pump unit 650 while the internal pressure is adjusted, thereby improving the safety.
  • the discrimination support hole 670 may be formed at a position adjacent to the discrimination pump unit 650 and the discrimination support hole 670 may be formed in the main body chip 650 when the discrimination pump unit 650 sucks air. 700 can be prevented from being deformed. That is, it is possible to prevent the mixed material from flowing out due to the deformation of the main body chip 700.
  • the main body chip 700 is provided inside the frame unit 800 to form a body, and the injected sample can be moved.
  • the main body chip 700 may include a main body upper film 710, a main body film 720, and a main body lower film 730, and a plurality of films may be stacked to form a channel.
  • the frame device 800 includes an upper frame 810 and a lower frame 820 to form an outer shape of the integrated automatic gene discrimination chip 1000. Specifically, the upper frame 810 and the lower frame 820 are coupled to each other to form an appearance of the integrated chip 1000 for an automatic gene discrimination.
  • the frame unit 800 may be made of a plastic material, but the material of the frame unit 800 is not limited thereto.
  • a plurality of alignment holes may be formed in the main body chip 700, and a columnar alignment column provided in the frame unit 800 may be inserted into the alignment holes.
  • the alignment pillars may be inserted into the alignment holes formed in the integrated automatic gene discrimination chip 1000 to fix the respective structures at predetermined positions.
  • the integrated automatic gene discrimination integrated chip 1000 is compact and convenient to carry. In addition, the integrated automatic gene discrimination chip 1000 is practical since it can be used immediately on the spot because the gene can be identified quickly. In addition, since the integrated chip 1000 for automatic gene discrimination does not need to fix the probe corresponding to the gene to the discriminator 600 in advance, it is possible to shorten the preparation time for gene discrimination and quickly identify the gene Do.
  • FIG. 10 is a flowchart of a gene discrimination method according to an embodiment of the present invention
  • FIG. 11 is a flowchart of a step of concentrating a detection target bacterium of a gene discrimination method according to an embodiment of the present invention.
  • the gene discrimination method can first perform step S100 of concentrating the detection target bacteria.
  • the step (S100) of concentrating the bacteria to be detected comprises a step (S110) of producing a primary mixed solution by mixing the desorption solution and the magnetic particles, a step of injecting the generated primary mixed solution into the concentration part (S120 A step S140 of discharging the foreign substance solution from the injected primary mixed solution to concentrate the detection target bacteria (S130), transferring the secondary mixed solution in the concentrated state to the extraction part (S140) And discharging the foreign substance solution from the secondary mixed solution to re-concentrate the detection target bacteria (S150).
  • FIG. 12 is a flowchart illustrating a step of generating a primary mixed solution in the step of concentrating a detection target bacterium in the gene discrimination method according to an embodiment of the present invention.
  • step S110 of mixing the desorption solution and the magnetic particles to form a primary mixed solution first, a step S111 of injecting a desorption solution and magnetic particles into the mixing vessel unit is performed .
  • a step S112 of heating the mixing vessel unit to a predetermined temperature may be performed.
  • the mixing vessel unit 111 can be heated to a temperature at which the mixed heating unit 112 can actively mix the desorption solution and the magnetic particles.
  • the mixing vessel unit 111 may be heated to 32 to 42 degrees.
  • vibration may be applied to the mixing vessel unit to mix the desorption solution and the magnetic particles to generate a primary mixed solution (S113).
  • the mixing vessel unit 111 is subjected to vibration for 20 to 120 minutes by the mixing motor unit 113, so that the desorption solution and the magnetic particles can be mixed.
  • the detection target microorganism and the magnetic particles may be combined.
  • FIG. 13 is a flowchart illustrating a step of injecting a primary mixed solution of a step of concentration of a detection target bacterium in a method for identifying a gene according to an embodiment of the present invention into a concentrating part.
  • the step of injecting the generated primary mixed solution into the concentrating unit (S120) may be performed by first performing a step S121 of generating a magnetic force in the concentrating container unit using the concentrated magnet unit have.
  • the concentrated magnet unit 142 is disposed adjacent to the concentrating container unit 141 by the concentrating motor unit 143 So that a magnetic force can be generated in the concentrating container unit 141.
  • step (S121) of generating a magnetic force to the concentrating container unit using the concentrated magnet unit when the concentrated magnet unit 142 is provided as an electromagnet a current is supplied to the concentrated magnet unit 142, It is also possible to generate a magnetic force in the magnet 141.
  • the step of injecting the primary mixed solution into the concentrating container unit using the concentrating pump unit (S122) may be performed.
  • the detection subject bacteria contained in the primary mixed solution may be attached to the inner wall of the concentrating container unit 141 by magnetic force in a state of being combined with the magnetic particles.
  • FIG. 14 is a flowchart illustrating a step of concentrating the detection target bacteria by discharging a foreign matter solution in the step of concentrating the detection target bacteria in the gene discrimination method according to an embodiment of the present invention.
  • the step S130 of discharging the foreign material solution from the injected primary mixed solution to concentrate the detection target bacteria is performed first (S131) of injecting the cleaning solution into the concentration portion .
  • the first discharge valve unit 152 is opened and the buffer unit 120 can inject the cleaning solution into the concentrating container unit 141.
  • the foreign substance solution may be discharged to the foreign substance discharging unit together with the washing solution to concentrate the detection target bacteria (S132).
  • the cleaning solution can transfer the foreign matter solution in the primary mixed solution contained in the concentrating container unit 141 to the foreign matter outlet unit 151 and discharge it.
  • the bacteria and the magnetic particles to be detected attached to the inner wall of the concentrating container unit 141 are not conveyed to the foreign matter outlet unit 151 together with the cleaning solution, but by the magnetic force of the concentrated magnet unit 142 By keeping the fixed state, it can be concentrated.
  • FIG. 15 is a flowchart illustrating a step of transferring a second mixed solution in the step of concentrating the detection target bacteria in the gene discrimination method according to an embodiment of the present invention to the extraction unit.
  • the magnetic force generated in the concentrating container unit is first removed, (S141) of generating a magnetic force in the unit.
  • the extraction magnet unit 182 may move adjacent to the extraction container unit 181 by the extraction motor unit 183 to generate a magnetic force in the extraction container unit 181.
  • the extraction magnet unit 182 in the step (S141) of removing magnetic force generated in the concentrating container unit and generating magnetic force in the extraction container unit using the extraction magnet unit when the extraction magnet unit 182 is provided as an electromagnet, the extraction magnet unit 182 to generate a magnetic force in the extraction container unit 181.
  • the concentrating unit 140 separates the concentrated magnet unit 142 from the concentrating container unit 141, or prevents current from flowing through the concentrated magnet unit 142, Can be removed.
  • step (S142) of generating vibration in the concentrating container unit can be performed.
  • the vibration motor unit 144 provided in the enrichment unit 140 generates vibration in the enrichment container unit 141 to remove the detection target bacteria attached to the enrichment container unit 141 . That is, the detection target bacteria adhering to the inner wall of the thickening part 140 can be more quickly transferred to the extracting part 180.
  • the transferring solution may be injected into the concentrating unit to transfer the secondary mixed solution to the extraction container unit (S143).
  • the buffer unit 120 injects the transfer solution into the concentrating unit 140 while the enrichment valve unit 145 is opened, thereby to transfer the secondary mixed solution to the extraction container unit 181 .
  • FIG. 16 is a flowchart showing a step of re-concentrating the detection target bacteria in the step of concentrating the detection target bacteria in the gene discrimination method according to an embodiment of the present invention.
  • step S150 of discharging the foreign matter solution from the transferred secondary mixed solution to re-concentrate the detection target bacteria first, the detection target bacteria combined with the magnetic particles are collected in the extraction container Unit (S151) attached to the unit.
  • the magnetic force generated in the concentrating container unit is removed, and the magnetic force applied to the extraction container unit 181 in the step (S 141) of generating magnetic force in the extraction container unit using the extraction magnet unit
  • the bacteria to be detected contained in the secondary mixed solution passing through the container unit 181 can be attached to the extraction container unit 181.
  • the foreign substance solution transferred together with the transferring solution may be discharged to the foreign substance discharging unit, and the step of re-concentrating the detection target bacteria (S152) may be performed.
  • the second discharge valve unit 153 is switched to the open state.
  • the foreign matter solution other than the detection target bacteria combined with the magnetic particles may be transferred to the foreign matter discharging unit 150 together with the transferring solution and discharged.
  • 17 is a flowchart of a step of extracting a gene of the gene discrimination method according to an embodiment of the present invention.
  • step (S200) of extracting genes from the concentrated detection subject bacteria can be performed.
  • (S200) of extracting a gene from the concentrated detection target bacteria is performed by injecting an extraction solution into an extraction unit containing the detection target bacteria combined with magnetic particles (S210), heating the extraction unit to a predetermined temperature, (S220) a step of extracting the gene from the target organism, and a step S230 of cooling the extracting unit to room temperature, and transferring the extracted gene toward the mixer (S240).
  • the extraction chamber unit 160 includes the extraction unit 180 ). ≪ / RTI >
  • the extraction solution may be a solution capable of extracting the gene in the bacteria to be detected by disrupting the bacteria to be detected.
  • FIG. 18 is a flowchart illustrating a step of extracting a gene from a detection subject bacterium in the step of extracting a gene of the gene discrimination method according to an embodiment of the present invention.
  • a step S220 of extracting a gene from the detection target bacteria by heating the extracting unit to a predetermined temperature includes firstly removing the magnetic force applied to the extracting unit by using the extracting magnet unit (S221).
  • the extraction magnet unit 182 is separated from the extraction container unit 181 and is connected to the extraction unit 180 The applied magnetic force can be removed.
  • the extracted magnet unit 182 is provided with an electromagnet
  • the current flowing in the extracted magnet unit 182 is controlled So that the magnetic force applied to the extraction unit can be removed.
  • the extracting unit may be heated to a predetermined temperature to extract a gene from the detection target organism (S222).
  • the extraction section 180 can be heated at 90 to 100 degrees for 5 to 20 minutes. If the temperature of the extraction container unit 181 is less than 90 degrees, the extraction solution may not actively active. If the temperature of the extraction container unit 181 exceeds 100 degrees, the gene may be destroyed, The function of the solution may be lost. Thus, the extraction container unit 181 can be heated from 90 to 100 degrees for 5 minutes to 20 minutes.
  • the temperature of the extraction container unit 181 is not limited to that described above, and it may be maintained at room temperature depending on the type of extraction solution.
  • the extraction solution contained in the extraction container unit 181 heated in an appropriate range can actively break down the cells of the detection target bacteria to rapidly extract genes.
  • the extracting unit After the extracting unit is heated to a preset temperature and the gene is extracted from the detection target bacteria (S220), the extracting unit may be cooled to room temperature (S230). At this stage, the extraction container unit 181 can be quickly cooled by 20 to 30 degrees by the extraction temperature control unit 184 and the extraction cooling unit 185.
  • FIG. 19 is a flowchart illustrating a step of transferring a gene to a blender in the step of extracting a gene of the gene discrimination method according to an embodiment of the present invention.
  • the step of transferring the extracted gene toward a blender may first perform step S241 of applying a magnetic force to the extracting unit.
  • the detection target bacteria contained in the extraction container unit 181 are in a state of being fixed to the inner wall of the extraction container unit 181 by the magnetic force because they are in a state of being coupled with the magnetic particles.
  • the gene extracted from the detection target bacteria is not fixed to the inner wall of the extraction container unit 181 because it is not coupled with the magnetic particles.
  • the step of transferring the gene extracted by the extracting unit toward the blender may be performed.
  • the extraction air injection unit 170 may open the extraction air injection valve 172 to inject air into the extraction unit 180.
  • the second discharge valve unit 153 is closed, and the mixer transfer valve unit 190 may be in an open state. In this state, the air injected into the extracting unit 180 may transfer the gene to the mixer 200.
  • the bacteria to be detected attached to the extraction container unit 181 and the magnetic particles are not transferred to the mixer 200, so they can be separated from the gene and discharged naturally.
  • the extraction chamber 160 further injects the extraction solution into the extraction unit 180 to smoothly transfer the gene .
  • FIG. 20 is a flowchart of a step of forming a mixed PCR solution of a gene discrimination method according to an embodiment of the present invention.
  • step S300 of forming a mixed PCR solution by mixing the extracted gene with the PCR buffer can be performed .
  • step S310 of injecting a PCR buffer into the main mixer chamber may be performed.
  • the PCR buffer is injected from the mixer main chamber extension 212 by the mixer pump unit 240 before the gene is introduced into the mixer main chamber body 211, To the front end of the housing.
  • a step S320 of injecting a gene into the mixer main chamber portion into which the PCR buffer is injected may be performed.
  • the gene injected through the mixer injection hole 213 may be mixed with the PCR buffer while bubbles are discharged by the mixer air filter 214 formed in the upper part of the mixer injection hole 213 .
  • a step S330 of injecting and sucking air such that the injected gene and the PCR buffer are mixed while reciprocating the mixer main chamber portion and the mixer auxiliary chamber portion can be performed.
  • the mixer pump unit 240 sucks the mixed PCR solution contained in the mixer main chamber unit 210 and moves the mixed PCR solution to the mixer auxiliary chamber unit 220, As shown in FIG. 7 (c), air may be injected into the mixed PCR solution contained in the mixer auxiliary chamber 220 and transferred to the mixer main chamber 210.
  • the mixer pump unit 240 may mix the gene and the PCR buffer by reciprocating the mixer auxiliary chamber unit 220 and the mixer main chamber unit 210 a plurality of times.
  • the gene and the PCR buffer when mixed while reciprocating between the mixer auxiliary chamber part 220 and the mixer main chamber part 210, they may collide with the vortex induction unit 215 to generate a vortex.
  • the resulting vortex can improve the mixing efficiency of the gene and the PCR buffer.
  • the mixed PCR solution in which the gene and the PCR buffer are mixed can be sucked and transferred to the mixer transfer chamber part (S340).
  • the mixer transfer chamber unit 230 can receive and receive the mixed PCR solution contained in the mixer auxiliary chamber unit 220 by the suction force of the mixer pump unit 240.
  • air may be injected into the mixer transfer chamber portion to transfer the mixed PCR solution to the quantitative injector (S350).
  • the mixer pump unit 240 may inject air to discharge the mixed PCR solution contained in the mixer transfer chamber unit 230 toward the metering injector 300.
  • Such a gene discrimination method improves the mixing efficiency of the gene and the PCR buffer, improves the reliability of the experimental results, and enables rapid experimentation.
  • FIG. 21 is a flowchart of a step of injecting a gene according to an embodiment of the present invention.
  • the mixed PCR solution thus formed may be quantitatively injected into the amplifier (S400) .
  • the step S410 of filling the mixed PCR solution sequentially from the quantification chamber located upstream may be performed. This step may be performed automatically by the quantitative stopper having a stepped portion through which the mixed PCR solution is passed after the mixed PCR solution is filled in the quantification chamber located upstream.
  • the introduced mixed PCR solution is first directed to the first quantitative stopper 321.
  • the first quantitative stopper 321 has a stepped height, the mixed PCR solution can not pass over the first quantitative stopper 321 and is naturally transferred to the first quantitative chamber 311, And is filled in the first metering chamber 311.
  • the first quantitative chamber 311 is filled with the mixed PCR solution, the water level of the mixed PCR solution rises by the step height of the first quantitative stopper 321, and the first quantitative stopper 321 It passes over.
  • the mixed PCR solution having passed through the first quantitative stopper 321 is directed to the second quantitative stopper 322.
  • the second quantitative stopper 322 also has a stepped height, the mixed PCR solution moves toward the second quantitative chamber 312 without passing over the second quantitative stopper 322, The chamber 312 is filled.
  • the second quantitative chamber 312 is filled with the mixed PCR solution, the water level of the mixed PCR solution rises by the step height of the second quantitative stopper 322, and the second quantitative stopper 322 It passes over.
  • the third PCR chamber 313 and the fourth PCR chamber 314 can be filled with the mixed PCR solution.
  • the quantitative stopper 320 is not provided in a stepped shape and the wall surface of the channel channel is made hydrophobic, the mixed PCR solution can be injected in a predetermined amount into each of the metering chambers in accordance with the difference in fluid resistance in the channel channel. .
  • a step S420 of filling the remaining portion of the metering chamber and discharging the remaining mixed PCR solution to the remaining amount discharging portion may be performed.
  • the discharge stopper 325 (see FIG. 3), which has a stepped portion to allow the excess mixed PCR solution to pass therethrough and be discharged to the remaining amount discharge portion 330 in a state in which the mixed PCR solution is filled in the dosing chamber portion 310 ).
  • the discharge stopper 325 which has a stepped portion to allow the excess mixed PCR solution to pass therethrough and be discharged to the remaining amount discharge portion 330 in a state in which the mixed PCR solution is filled in the dosing chamber portion 310 ).
  • step S430 of closing the metering valve unit may be performed.
  • the quantitative pump part 340 injects the mixed PCR solution stored in the quantification chamber part 310 into the amplifier (400).
  • the quantitative air filter unit 370 can remove bubbles contained in the mixed PCR solution transferred toward the amplifier 400.
  • the gene discrimination method as described above can be always injected into the amplifier 400 by a predetermined amount, thereby improving the reliability of the experimental results and automatically injecting the quantitation without any additional control, The experiment can be made possible.
  • PCR buffer for gene amplification may be lyophilized in a gene amplification chamber and loaded in advance.
  • amplified genes are amplified by polymerase chain reaction (PCR) on the quantified mixed PCR solution to form an amplification solution (S500).
  • PCR polymerase chain reaction
  • the mixed PCR solution injected in the quantitative manner is amplified by the amplification of the gene Can be amplified.
  • the gene can be amplified by polymerase chain reaction (PCR).
  • PCR polymerase chain reaction
  • FIG. 22 is a flowchart of a step of forming a mixed material of the gene discrimination method according to an embodiment of the present invention.
  • an amplification solution and a signal substance are mixed
  • Forming a mixed material may be performed.
  • step S600 of forming the mixed material by mixing the amplification solution and the signal material a step (S611) of injecting the amplification solution into the mixing chamber part into which the signal material is injected, and a step of injecting the amplification solution and the signal material into the mixing chamber part (S612) of injecting and sucking air to be mixed while reciprocating the mixed auxiliary chamber portion.
  • the signal material is injected into the mixing chamber chamber extension 512 (FIG. 5) before the amplification solution is introduced into the mixing chamber chamber body 511, at step S611, And may be filled up to the front end of the mixed injection hole 513.
  • the amplification solution injected through the mixing hole 513 is mixed with the mixture
  • the air bubbles can be mixed with the signal material while being discharged by the mixed air filter 514 formed in the upper portion of the injection hole 513.
  • the discrimination pump part 650 is disposed in the mixing chamber part 510 Air can be sucked in and the signal material and the amplification solution can be transferred to the mixing auxiliary chamber part 520.
  • the discrimination pump unit 650 may inject air and then transfer the signal material and the amplification solution contained in the mixing auxiliary chamber unit 520 to the mixing chamber chamber unit 510. In this way, the discrimination pump unit 650 can mix the amplification solution and the signal material between the mixing auxiliary chamber part 520 and the mixing chamber part 510 a plurality of times.
  • the amplification solution and the signal material when the amplification solution and the signal material are mixed while reciprocating between the mixing auxiliary chamber part 520 and the mixing chamber part 510, the amplification solution and the signal material may collide with the vortex induction unit to generate a vortex.
  • the vortex generated as described above can improve the mixing efficiency of the amplification solution and the signal material.
  • FIG. 23 is a flowchart of a step of discriminating genes of a gene discrimination method according to an embodiment of the present invention.
  • a step S700 of determining the gene by measuring the amount of current of the mixed material may be performed .
  • Step S700 of determining the gene by measuring the amount of current of the mixed material formed may include applying an applied voltage to the working electrode (S710), measuring the electron derived from the redox reaction of the signal material included in the mixed material A step S720 of continuously exchanging the working electrode with the working electrode through an amount of electrons exchanged with the counter electrode S730 and a step S740 of discriminating the gene by analyzing the measured amount of current .
  • a step of applying an applied voltage to the working electrode S710 is a step of applying an applied voltage to the working electrode 611.
  • the applied voltage means a unique voltage value for causing a redox reaction depending on each signal material included in the mixed material. That is, by applying the applied voltage to the working electrode 611 in consideration of the reference voltage of the reference electrode 613, the redox reaction can be performed on the signal material contained in the mixed material.
  • the counter electrode 612 may oxidize the signal material contained in the mixed material Electrons derived through the reduction reaction can be exchanged with the working electrode 611.
  • the signal material causing the redox reaction may be a signal material Quot; Specifically, the binding amount of the signal material is determined according to the amount of the gene, and the signal material that is not coupled to the gene is extracted through the redox reaction to provide an electrical signal to the working electrode 611 .
  • the working electrode 611 sequentially changes the amount of current through the amount of electrons exchanged with the counter electrode 612 . That is, in the step S730 of continuously measuring the amount of current through the amount of electrons exchanged with the counter electrode by the working electrode, it is possible to successively measure the amount of current change according to the amount of the signal material not coupled to the gene .
  • step S740 of determining the gene by analyzing the measured amount of electric current when the amount of current is higher than a preset reference value, it is determined that the gene is less than the reference amount. If the amount of current is lower than the predetermined reference value, Many can be judged.
  • a food-borne pathogen discriminator using a gene discrimination method using a gene discrimination chip if the electric current is higher than a preset reference value, it is judged that the gene causing food poisoning is small and the food is judged to be safe. On the other hand, if the amount of electric current is lower than a preset reference value, it is judged that there are many genes causing food poisoning, and it is judged that there is a risk of food poisoning if the food is consumed.
  • the method for determining the gene is not limited to the food poisoning bacteria determination device.
  • the method of gene identification can be applied to a fish species discriminator.

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Abstract

La présente invention concerne une puce intégrée de lecture de gène entièrement automatique et, plus particulièrement, une puce intégrée de lecture de gène entièrement automatique conçue pour lire rapidement un gène de bactéries à détecter, de manière entièrement automatisée. La configuration de la présente invention concerne une puce intégrée de lecture de gène entièrement automatique comprenant : un extracteur de concentration pour enrichir des bactéries à détecter par décharge d'une solution de matière étrangère composée d'une solution combinée dans laquelle une solution de détachement et des particules magnétiques sont combinées, et pour extraire un gène des bactéries enrichies à détecter ; un mélangeur, disposé en aval de l'extracteur de concentration, pour mélanger le gène et un tampon de PCR pour former une solution de PCR mélangée ; un amplificateur, disposé en aval du mélangeur, pour effectuer une réaction en chaîne de polymérase (PCR) avec la solution de PCR mélangée pour donner une solution amplifiée ; un injecteur quantitatif, disposé entre le mélangeur et l'amplificateur, pour injecter une quantité déterminée de la solution de PCR mélangée à l'amplificateur ; un combineur, disposé en aval de l'amplificateur, pour combiner une substance de signalisation avec la solution d'amplification amplifiée par l'amplificateur pour former une matière combinée ; et un lecteur, disposé en aval du combineur, pour lire le gène contenu dans la matière combinée, le lecteur mesurant une quantité actuelle de la matière combinée pour lire le gène.
PCT/KR2017/011246 2017-07-24 2017-10-12 Puce intégrée de lecture de gène entièrement automatique WO2019022299A2 (fr)

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