WO2023171888A1 - Gene amplification and detection device using isothermal amplification and lateral flow assay - Google Patents

Gene amplification and detection device using isothermal amplification and lateral flow assay Download PDF

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Publication number
WO2023171888A1
WO2023171888A1 PCT/KR2022/020560 KR2022020560W WO2023171888A1 WO 2023171888 A1 WO2023171888 A1 WO 2023171888A1 KR 2022020560 W KR2022020560 W KR 2022020560W WO 2023171888 A1 WO2023171888 A1 WO 2023171888A1
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cap
tube
isothermal
amplification
reaction solution
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PCT/KR2022/020560
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French (fr)
Korean (ko)
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윤현규
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아토플렉스 주식회사
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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
    • B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
    • B01L3/508—Containers for the purpose of retaining a material to be analysed, e.g. test tubes rigid containers not provided for above
    • B01L3/5082—Test tubes per se
    • 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
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2200/00—Solutions for specific problems relating to chemical or physical laboratory apparatus
    • B01L2200/16—Reagents, handling or storing thereof
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2300/00—Additional constructional details
    • B01L2300/04—Closures and closing means
    • B01L2300/041—Connecting closures to device or container
    • B01L2300/042—Caps; Plugs
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2300/00—Additional constructional details
    • B01L2300/04—Closures and closing means
    • B01L2300/041—Connecting closures to device or container
    • B01L2300/044—Connecting closures to device or container pierceable, e.g. films, membranes
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2300/00—Additional constructional details
    • B01L2300/06—Auxiliary integrated devices, integrated components
    • B01L2300/0672—Integrated piercing tool
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2300/00—Additional constructional details
    • B01L2300/06—Auxiliary integrated devices, integrated components
    • B01L2300/069—Absorbents; Gels to retain a fluid
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2300/00—Additional constructional details
    • B01L2300/18—Means for temperature control
    • B01L2300/1805—Conductive heating, heat from thermostatted solids is conducted to receptacles, e.g. heating plates, blocks
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2400/00—Moving or stopping fluids
    • B01L2400/04—Moving fluids with specific forces or mechanical means
    • B01L2400/0403—Moving fluids with specific forces or mechanical means specific forces
    • B01L2400/0406—Moving fluids with specific forces or mechanical means specific forces capillary forces
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q2527/00—Reactions demanding special reaction conditions
    • C12Q2527/101—Temperature

Definitions

  • the present invention relates to a gene amplification and detection device, and in particular, to a device for amplifying and analyzing genes by combining isothermal amplification method and lateral flow analysis method without using a pipette.
  • PCR amplification method has been mainly used as a gene amplification method for detecting specific genes.
  • the PCR amplification method is a method of repeating the cycle of changing temperature dozens of times through three stages of gene amplification: denaturation, annealing, and extension. Therefore, temperature control is necessary and is expensive. It requires a lot of equipment and has the disadvantage of taking a lot of time to amplify the gene.
  • the isothermal amplification method using an isothermal amplifier has recently been used for rapid genetic testing.
  • An isothermal amplifier is a device that maintains a constant temperature of approximately 60°C to 65°C. Genetic samples and reaction materials for gene amplification are placed in a tube, and the tube is mounted on an isothermal amplifier to maintain a constant temperature of approximately 60°C to 65°C. When left for a certain period of time, gene amplification occurs within the tube.
  • the lateral flow analyzer is generally used in the widely known lateral flow assay (LFA). For example, it is widely used in pregnancy diagnosis devices to check whether pregnancy is present, and is also used to detect specific genes. .
  • LFA lateral flow assay
  • gene amplification is performed by labeling an antigen to a primer or probe used in the gene amplification process, and then the amplification product (labeled with the antigen) is spread on a membrane for lateral flow analysis, and the developed amplification It is used in a molecular genetic analysis method to detect the presence of a specific gene in an amplification product by determining whether the antigen of the product reacts with the antibody present on the membrane.
  • the gene amplification process and the gene detection process must be performed in separate devices.
  • the gene amplification product sample to be measured
  • the gene amplification product is transferred from a tube mounted on the isothermal amplifier to a lateral flow analyzer to determine the presence or absence of a specific gene. must be detected.
  • the technical field related to gene amplification and detection devices to which the present invention belongs also requires constant technological improvement.
  • the present invention was designed to solve the problems described above, and can detect target genes quickly and easily using a combination of isothermal amplification method and lateral flow analysis method.
  • the purpose is to provide a gene amplification and detection device that can obtain accurate and reliable detection results without being affected.
  • the present invention to achieve the above object is a gene amplification and detection device using isothermal amplification and lateral flow analysis
  • a reaction solution tube having an opening at the top, containing a reaction solution in the inner space, and in which a genetic sample is mixed and dissolved in the reaction solution;
  • a reaction material for gene amplification is accommodated in the inner space, an upper wall dividing the upper side of the inner space is provided with a cutout, and a lower side of the inner space is provided.
  • the bottom wall dividing the rupture line is provided with a rupture line that is ruptured by an external force, and when the rupture line is ruptured by an external force, the gene amplification reactant is discharged and mixed into the reaction solution contained in the inner space of the reaction solution tube.
  • reaction solution tube to which the tube cap is coupled is accommodated, and the received reaction solution tube is maintained at a constant temperature to react the mixture of the genetic sample, the reaction solution, and the gene amplification reactant in the inner space of the reaction solution tube.
  • an isothermal amplifier for isothermal amplification of the genetic sample isothermal amplification of the genetic sample
  • a lateral flow analyzer including a membrane provided along the longitudinal direction inside the casing and exposed to the inner space of the reaction solution tube through an opening at the lower tip and coming into contact with the isothermally amplified genetic sample within the reaction solution tube.
  • the tube cap includes an upper cap having the upper wall, a side wall dividing the internal space, and a lower opening, and the bottom wall. and a lower cap that has a side wall and the upper cap is coupled to an inner space defined by the side wall, and when the upper cap is pressed into the inner space of the lower cap, the bottom wall of the lower cap is connected to the upper cap. It may have a rupture line that ruptures when pressed by the bottom of the cap.
  • the rupture line of the bottom wall may be formed in a cross shape.
  • the side wall height of the upper cap is higher than the side wall height of the lower cap, and the lower end of the side wall of the upper cap is the bottom wall of the lower cap.
  • the isothermal amplifier includes an external cylinder, an isothermal chamber provided inside the external cylinder to accommodate the reaction liquid tube, and the isothermal chamber. It may be provided with a heat transfer plate that transfers heat to maintain a constant temperature in the reaction liquid tube contained in and a temperature control PCB board that can control the temperature of the heat transfer plate.
  • the upper cap may be made of polyethylene material, and the lower cap may be made of silicone rubber.
  • the reaction solution may include potassium hydroxide solution and polyethylene glycol
  • the gene amplification reactants include Bst DNA polymerase, It may contain buffer solution, primers, dNTP and Mg 2+ .
  • gene detection results can be obtained quickly and easily by using a combination of the isothermal amplification method and the lateral flow analysis method.
  • the reaction solution in which the gene sample in the reaction solution tube is dissolved and the reaction material for gene amplification in the tube cap are stored in the reaction solution tube. Can be easily mixed.
  • the assembly of the tube cap and the reaction tube is inserted into the isothermal chamber of the isothermal amplifier to isothermally amplify the genetic sample, and then the opening of the lateral flow analyzer where the membrane is exposed is inserted through the incision in the tube cap.
  • the isothermal amplification product in the reaction tube comes into contact with the membrane and flows along the membrane by capillary action, thereby visually indicating the presence or absence of the gene to be detected. Therefore, the present invention can quickly and easily perform gene amplification and detection using isothermal amplification and lateral flow analysis without using a pipette and without external influence.
  • the present invention can detect the target gene by simply combining a lateral flow analyzer with the isothermal amplifier from which the isothermal amplification product is obtained, thereby preventing contamination of the analyte and obtaining highly reliable results.
  • the assembly of the reaction tube and tube cap and the lateral flow analyzer can be easily separated from the isothermal amplifier, making it easy to remove after use and preventing contamination of the isothermal amplifier. There is an advantage.
  • Figure 1 is an exploded perspective view showing the parts constituting the gene amplification and detection device according to an embodiment of the present invention separated.
  • Figure 2 is a combined cross-sectional view of a gene amplification and detection device according to an embodiment of the present invention.
  • Figure 3a is a perspective view of a reaction liquid tube sealed with a film according to an embodiment of the present invention.
  • Figure 3b is a cross-sectional view of Figure 3a.
  • Figure 4A is a perspective view of a tube cap according to one embodiment of the present invention.
  • 4B is a bottom perspective view of a tube cap according to one embodiment of the present invention.
  • Figure 4c is a cross-sectional view of the tube cap taken along line 'I-I' of Figure 4a.
  • Figure 5a is a cross-sectional view of a tube cap coupled to a reaction liquid tube according to an embodiment of the present invention.
  • Figure 5b is a cross-sectional view of the tube cap in the state of Figure 5a when the upper cap of the tube cap is pressed and pressed into the lower cap.
  • Figure 6a is a cross-sectional view of the assembly of the tube cap and the reaction liquid tube before inserting it into the isothermal chamber of the isothermal amplifier according to one embodiment of the present invention.
  • Figure 6b is a cross-sectional view of the assembly of the tube cap and the reaction tube being inserted into the isothermal chamber of the isothermal amplifier and performing isothermal amplification according to an embodiment of the present invention.
  • Figure 7a is a perspective view of a lateral flow analyzer according to an embodiment of the present invention.
  • Figure 7b is a cross-sectional view of a lateral flow analyzer according to an embodiment of the present invention.
  • the gene amplification and detection device using an isothermal amplifier and a lateral flow analyzer has an opening at the top and contains a reaction solution (L) (gene a reaction liquid tube 10 that accommodates the sample (the sample is mixed and dissolved), a tube cap 20 that is coupled to the opening of the reaction liquid tube 10 and seals the inside of the reaction liquid tube 10, and a tube cap.
  • L reaction solution
  • reaction solution tube 10 accommodates the combined reaction solution tube 10 and maintains an isothermal state to isothermally amplify the gene sample in the reaction solution tube 10, an isothermal amplifier 30, a tube cap 20, and a reaction solution tube ( It includes a lateral flow analyzer (40) that is inserted into the assembly of 10) and detects a specific target gene by contacting the isothermal amplification product in the reaction tube (10) amplified in the isothermal amplifier (30) with the membrane (47). .
  • a lateral flow analyzer 40
  • the reaction liquid tube 10 accommodates the reaction liquid (L) in its internal space, and is sealed with a film 15 attached to the opening 11 at the top before use. The inside is blocked from the outside.
  • the user inserts a cotton swab (not shown) into the nose and rubs the inside of the nose to collect a sample on the swab.
  • the cotton swab with the sample is placed in the reaction liquid tube 10 ( When placed in L), the genetic sample from the sample on the cotton swab is dissolved in the reaction solution (L).
  • the reaction solution (L) contained in the reaction solution tube 10 contains potassium hydroxide solution and polyethylene glycol, and dissolves the genetic sample from the specimen affixed to the cotton swab.
  • the tube cap 20 is coupled to the opening 11 of the reaction liquid tube 10.
  • the reaction solution tube 10 may be made of, for example, polypropylene, but is not limited thereto, and of course, various materials suitable for stable reaction solution storage and gene amplification may be used.
  • the tube cap 20 shown in FIGS. 4A to 4C consists of an upper cap 21 and a lower cap 26.
  • the upper cap 21 has a cutout 23 formed on the upper wall, has a side wall that partitions the internal space S 20 , and the lower end is open to form an opening.
  • the lower cap 26 has an inner space formed by the side wall and the upper end is open to form an opening.
  • the side wall of the upper cap 21 is movably fitted and coupled into the inner space of the lower cap 26 through the upper opening of the lower cap 26.
  • a 'cross (+)' shaped rupture line 27 is formed on the bottom wall 28 of the lower end of the lower cap 20.
  • the reaction material (p) for gene amplification is accommodated in the internal space (S 20 ) of the upper cap (21).
  • the reactant for gene amplification (p) can be freeze-dried, and the freeze-dried reactant for gene amplification (p) contains Bst DNA polymerase, buffer solution, primers, dNTP, and Mg 2+ required for isothermal amplification of gene samples. Includes.
  • the side wall height h1 of the upper cap 21 is higher than the side wall height h2 of the lower cap 26. Therefore, when the upper cap 21 is coupled to the lower cap 26 with the lower end of the side wall of the upper cap 21 in contact with the bottom wall 28 of the lower cap 26, the upper and lower sides of the upper cap 21 A gap D is formed between the tops of the caps 26.
  • the upper cap 21 may be made of polyethylene material
  • the lower cap 26 may be made of silicone rubber.
  • the upper cap 21 and the lower cap 26 are not limited to the above-described materials, and provide stable storage of reactants for gene amplification and efficient removal and dropping of reactants for gene amplification when combined with the reaction tube 10. , and of course, various materials suitable for gene amplification can be used.
  • the tube cap 20 with the upper cap 21 and lower cap 26 assembled is the opening of the reaction liquid tube 10 from which the vinyl 15 has been removed. It is coupled to (11). Accordingly, the inner space of the reaction solution tube (10) containing the reaction solution in which the genetic sample is dissolved is sealed, and foreign substances are prevented from entering the internal space.
  • the bottom wall 28 is divided while being elastically pressed by the lower side of the side wall of the upper cap 21, and the elastic divided The bottom wall 28 is tightly fitted between the inside of the side wall of the reaction liquid tube 10 and the side wall of the upper cap 21.
  • the mixture of the reaction solution (L) in which the genetic sample is dissolved and the reaction material for gene amplification (p) is kept at a constant temperature, e.g. When reacted at 60°C to 65°C for approximately 30 minutes, the gene to be detected is isothermally amplified.
  • the isothermal amplifier 30 includes an external cylinder 31, an isothermal chamber 32 provided inside the external cylinder 31 to accommodate the reaction tube 10, and an isothermal chamber 32.
  • a heat transfer plate 34 is provided on at least one of the side walls and the bottom of the isothermal chamber 32 to transfer heat to the reaction liquid tube 10 accommodated in the isothermal chamber 32 to maintain a constant temperature, and a heat transfer plate is installed on the bottom of the isothermal chamber 32.
  • It includes a temperature control PCB board (35) capable of controlling the temperature (34).
  • the reaction liquid tube 10 coupled with the tube cap 20 is inserted into the isothermal chamber 32 of the isothermal amplifier 30, and the temperature control PCB board 35 is operated.
  • the heating plate 34 is heated and the isothermal chamber 32 of the isothermal amplifier 30 is maintained at a constant temperature, for example, 60° C. to 65° C. for approximately 30 minutes, the reaction liquid (L) in the reaction liquid tube 10 ) reacts with the gene amplification reactant (p) (including Bst DNA polymerase, buffer solution, primers, dNTP, and Mg 2+ ), and the gene to be detected is isothermally amplified.
  • p gene amplification reactant
  • the lateral flow analyzer 40 is formed in the casing 41 and the lower part of the casing 41, and is coupled to the assembly of the reaction liquid tube 10 and the tube cap 20. When doing so, it penetrates the space between the cutout part 23 of the tube cap 20 and the quartered bottom wall 28 and is inserted into the inner space of the reaction liquid tube 10, and has an opening 44 on the bottom surface.
  • a vibrating lower tip 43 and the casing 41 are provided along the longitudinal direction, and the lower end is exposed to the inner space of the reaction liquid tube 10 through the opening 44 of the lower tip 43 to react. It is provided with a membrane 47 that comes into contact with the isothermally amplified gene sample (liquid sample) in the liquid tube 10.
  • the casing 41 consists of a left casing part 41a and a right casing part 41b, and a membrane 47 is provided therein in the longitudinal direction.
  • the membrane 47 is composed of a porous membrane through which the isothermally amplified gene sample (liquid sample) can flow through capillary action.
  • an absorption pad 112 that promotes capillary action
  • a lateral flow analyzer (40) is inserted into the incision (23) provided on the upper wall of the upper cap (21) of the assembly of the reaction liquid tube (10) and the tube cap (20) accommodated in the isothermal chamber (32) of the isothermal amplifier (30). ), the lower tip 43 of the lateral flow analyzer 40 penetrates the cutout 23 of the upper cap 21 and separates the upper cap 21 and the lower cap. It sequentially passes through (26) and enters the reaction solution tube 10 to be immersed in the isothermally amplified gene sample (liquid sample) in the reaction solution tube 10 (see FIG. 2).
  • the isothermally amplified gene sample (liquid sample) in the reaction tube 10 comes into contact with the surface of the membrane 47 through the opening 44 of the lower tip 43 of the lateral flow analyzer 40. , Afterwards, the gene sample (liquid sample) amplified isothermally by capillary action flows smoothly along the membrane 47. If a gene to be detected is present in the flowing liquid sample, a band appears in the detection area of the membrane 47. This visually shows whether a gene to be detected, for example, a specific viral gene, is present in a sample collected from the nose, and users or medical staff can easily check whether they are infected with a specific virus. Therefore, according to the present invention, gene amplification and detection using isothermal amplification and lateral flow analysis can be performed quickly and easily without using a pipette and without external influence.
  • the lateral flow analyzer 40 coupled to the assembly of the reaction tube 10 and the tube cap 20 is separated from the isothermal amplifier 30. do. Since the lateral flow analyzer 40 is separated from the isothermal amplifier 30 while being coupled to the assembly of the reaction tube 10 and the tube cap 20, it is convenient to separate and dispose of the analysis device after the test is completed. Therefore, the present invention has the advantage of being easy to remove after use and preventing contamination of the isothermal amplifier.

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  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Clinical Laboratory Science (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Analytical Chemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Hematology (AREA)
  • Apparatus Associated With Microorganisms And Enzymes (AREA)
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Abstract

The present invention provides a gene amplification and detection device using isothermal amplification and a lateral flow assay, wherein a target gene can be promptly and conveniently detected by using a combination of an isothermal application method and a lateral flow assay and an accurate detection result with high reliability can be obtained by combining an isothermal amplifier and a lateral flow analyzer, without influences from the outside.

Description

등온증폭 및 측방유동분석을 이용한 유전자 증폭 및 검출장치Gene amplification and detection device using isothermal amplification and lateral flow analysis
본 발명은 유전자 증폭 및 검출장치에 관한 것으로서, 특히 피펫을 사용하지 않고, 등온증폭방법과 측방유동분석법을 조합하여 유전자를 증폭 및 분석하는 장치에 관한 것이다. The present invention relates to a gene amplification and detection device, and in particular, to a device for amplifying and analyzing genes by combining isothermal amplification method and lateral flow analysis method without using a pipette.
일반적으로 특정 유전자 검출을 위한 유전자 증폭방식으로서 PCR 증폭방식을 주로 사용해 오고 있다. PCR 증폭방식은 유전자 증폭 시 변성(denaturation), 어닐링(annealing) 및 연장(extension)의 세 가지 단계를 거치면서 온도의 변화를 주는 사이클을 수십회 반복하는 방식이기 때문에 온도제어가 필요하고 이를 위한 고가의 장비가 필요하며 유전자 증폭에 시간이 많이 소요되는 단점을 가지고 있다.In general, PCR amplification method has been mainly used as a gene amplification method for detecting specific genes. The PCR amplification method is a method of repeating the cycle of changing temperature dozens of times through three stages of gene amplification: denaturation, annealing, and extension. Therefore, temperature control is necessary and is expensive. It requires a lot of equipment and has the disadvantage of taking a lot of time to amplify the gene.
유전자 증폭기술로서 상기한 단점을 가진 PCR 증폭방식 대신에 최근 들어 등온증폭기를 이용한 등온증폭방법이 신속 유전자 검사를 위해 사용되고 있다. 등온증폭기는 대략 60℃ 내지 65℃의 일정한 온도를 유지시켜 주는 장치로서, 유전자 시료와 유전자 증폭용 반응물질을 튜브에 넣고, 상기 튜브를 등온증폭기에 장착한 후 대략 60℃ 내지 65℃의 일정한 온도에서 일정 시간 동안 두게 되면 상기 튜브 내에서 유전자 증폭이 이루어진다. As a gene amplification technology, instead of the PCR amplification method, which has the above-mentioned disadvantages, the isothermal amplification method using an isothermal amplifier has recently been used for rapid genetic testing. An isothermal amplifier is a device that maintains a constant temperature of approximately 60°C to 65°C. Genetic samples and reaction materials for gene amplification are placed in a tube, and the tube is mounted on an isothermal amplifier to maintain a constant temperature of approximately 60°C to 65°C. When left for a certain period of time, gene amplification occurs within the tube.
측방 유동분석기는 일반적으로 널리 알려진 측방유동분석법(Lateral Flow Assay: LFA)에 사용되는 것으로서, 예를 들어 임신의 여부를 확인할 수 있는 임신진단장치 등에 널리 이용되고 있고, 특정 유전자 검출을 위해서도 활용되고 있다. The lateral flow analyzer is generally used in the widely known lateral flow assay (LFA). For example, it is widely used in pregnancy diagnosis devices to check whether pregnancy is present, and is also used to detect specific genes. .
측방유동분석법은 특정 유전자 검출을 위해 유전자 증폭 과정에 사용되는 프라이머 또는 프로브에 항원을 표지하여 유전자 증폭을 수행한 후 증폭산물(항원이 표지됨)을 측방 유동분석용 멤브레인에 전개시키고, 전개된 증폭산물의 항원이 멤브레인 상에 존재하는 항체와의 반응여부로 증폭 산물 내에 특정 유전자의 존재 여부를 검출하는 분자유전학적 분석법에 이용된다.In the lateral flow analysis method, to detect a specific gene, gene amplification is performed by labeling an antigen to a primer or probe used in the gene amplification process, and then the amplification product (labeled with the antigen) is spread on a membrane for lateral flow analysis, and the developed amplification It is used in a molecular genetic analysis method to detect the presence of a specific gene in an amplification product by determining whether the antigen of the product reacts with the antibody present on the membrane.
그런데 상기한 바와 같이 등온증폭방법과 측방유동분석법을 함께 이용할 경우, 유전자 증폭과정과 유전자 검출과정을 각각 다른 장치에서 수행해야 하는 문제가 있다. 즉, 등온증폭기에서 유전자 증폭과정을 수행한 후, 피펫과 같은 별도의 도구를 사용하여, 유전자 증폭산물(피측정 시료)를 등온증폭기에 장착된 튜브로부터 측방유동분석기로 옮겨서 특정 유전자의 존재 유무를 검출해야 한다. 이러한 방식으로 유전자 증폭 및 분석을 할 경우, 피측정 시료를 하나의 장치에서 다른 장치로 옮기는 과정이 불편할 뿐만아니라 이러한 과정에서 피측정 시료가 오염될 우려가 있고, 피펫 팁 및 튜브의 폐기가 불편하며 등온증폭기에 대한 오염의 우려가 있다.However, as described above, when using the isothermal amplification method and the lateral flow analysis method together, there is a problem that the gene amplification process and the gene detection process must be performed in separate devices. In other words, after performing the gene amplification process in an isothermal amplifier, using a separate tool such as a pipette, the gene amplification product (sample to be measured) is transferred from a tube mounted on the isothermal amplifier to a lateral flow analyzer to determine the presence or absence of a specific gene. must be detected. When gene amplification and analysis are performed in this way, not only is it inconvenient to transfer the sample to be measured from one device to another, but there is also a risk of contamination of the sample to be measured in this process, and disposal of pipette tips and tubes is inconvenient. There is a risk of contamination of the isothermal amplifier.
따라서, 본 발명이 속한 유전자 증폭 및 검출장치와 관련한 기술분야 역시 다른 기술분야와 마찬가지로 끊임없는 기술의 개선이 필요하다. Therefore, like other technical fields, the technical field related to gene amplification and detection devices to which the present invention belongs also requires constant technological improvement.
한편, 상기한 배경기술로서 설명된 사항들은 본 발명의 배경에 대한 이해 증진을 위한 것일 뿐, 본 발명의 "선행 기술"로서 이용될 수 있다는 승인으로서 인용한 것은 아님을 이해하여야 한다.Meanwhile, it should be understood that the matters described as the above-mentioned background art are only for the purpose of improving understanding of the background of the present invention, and are not cited as approval that they can be used as “prior art” of the present invention.
본 발명은 상기한 바와 같은 문제점을 해결하기 위하여 고안된 것으로서, 등온증폭방법 및 측방유동분석법의 조합을 사용하여 신속하고 간편하게 표적 유전자를 검출할 수 있고, 등온증폭기와 측방유동분석기를 하나로 결합하여 외부의 영향을 받지 않으면서도 정확하고 신뢰성이 높은 검출결과를 얻을 수 있는 유전자 증폭 및 검출장치를 제공함에 그 목적이 있다. The present invention was designed to solve the problems described above, and can detect target genes quickly and easily using a combination of isothermal amplification method and lateral flow analysis method. By combining the isothermal amplifier and lateral flow analyzer into one, external The purpose is to provide a gene amplification and detection device that can obtain accurate and reliable detection results without being affected.
그러나, 전술한 바와 같은 본 발명의 과제는 예시적인 것으로, 이에 의해 본 발명의 범위가 제한되는 것은 아니다. 또한, 본 발명의 다른 목적 및 이점은 하기의 발명의 상세한 설명, 청구범위 및 도면에 의해 보다 명확하게 된다.However, the object of the present invention as described above is illustrative, and the scope of the present invention is not limited thereby. Additionally, other objects and advantages of the present invention will become clearer from the following detailed description, claims, and drawings.
상기 목적을 달성하기 위한 본 발명은 등온증폭 및 측방유동분석을 이용한 유전자 증폭 및 검출장치로서, The present invention to achieve the above object is a gene amplification and detection device using isothermal amplification and lateral flow analysis,
상단에 개방부를 구비하고, 내부공간에는 반응액이 수용되어 있으며, 유전자 시료가 상기 반응액에 혼합되어 용해되는 반응액 튜브와,A reaction solution tube having an opening at the top, containing a reaction solution in the inner space, and in which a genetic sample is mixed and dissolved in the reaction solution;
상기 반응액 튜브의 개방부를 밀폐하도록 상기 개방부에 결합되고, 내부 공간에는 유전자 증폭용 반응물질이 수용되어 있으며, 상기 내부 공간의 상측을 구획하는 상단 벽에는 절개부를 구비하고, 상기 내부 공간의 하측을 구획하는 바닥벽에는 외력에 의해 파열되는 파열선을 구비하며, 상기 파열선이 외력에 의해 파열될 때 상기 유전자 증폭용 반응물질이 상기 반응액 튜브의 내부공간에 수용된 반응액으로 배출되어 혼합되는 튜브 캡과,It is coupled to the opening to seal the opening of the reaction solution tube, a reaction material for gene amplification is accommodated in the inner space, an upper wall dividing the upper side of the inner space is provided with a cutout, and a lower side of the inner space is provided. The bottom wall dividing the rupture line is provided with a rupture line that is ruptured by an external force, and when the rupture line is ruptured by an external force, the gene amplification reactant is discharged and mixed into the reaction solution contained in the inner space of the reaction solution tube. tube cap,
상기 튜브 캡이 결합된 반응액 튜브를 수용하고, 상기 수용된 반응액 튜브를 일정한 온도에서 유지하여 상기 반응액 튜브의 내부공간 내의 상기 유전자 시료, 상기 반응액 및 상기 유전자 증폭용 반응물질의 혼합액을 반응시켜 상기 유전자 시료를 등온증폭시키는 등온증폭기와,The reaction solution tube to which the tube cap is coupled is accommodated, and the received reaction solution tube is maintained at a constant temperature to react the mixture of the genetic sample, the reaction solution, and the gene amplification reactant in the inner space of the reaction solution tube. an isothermal amplifier for isothermal amplification of the genetic sample,
케이싱; 상기 케이싱의 하부에 형성되고, 하단에는 개방된 개구부를 가지며, 상기 튜브 캡의 절개부와 바닥벽을 관통하여 상기 반응액 튜브의 내부공간으로 삽입되는 하부 첨단부; 및 상기 케이싱 내부에 길이방향을 따라 제공되고, 상기 하부 첨단부의 개구부를 통하여 상기 반응액 튜브의 내부공간에 노출되어 상기 반응액 튜브 내의 등온증폭된 유전자 시료와 접촉하게 되는 멤브레인을 구비하는 측방유동분석기를 포함한다. casing; a lower tip formed at the lower part of the casing, having an open opening at the lower end, and inserted into the inner space of the reaction liquid tube through the cutout and bottom wall of the tube cap; and a lateral flow analyzer including a membrane provided along the longitudinal direction inside the casing and exposed to the inner space of the reaction solution tube through an opening at the lower tip and coming into contact with the isothermally amplified genetic sample within the reaction solution tube. Includes.
본 발명의 일 구체예의 등온증폭 및 측방유동분석을 이용한 유전자 증폭 및 검출장치에 있어서, 상기 튜브 캡은 상기 상단 벽, 상기 내부공간을 구획하는 측벽 및 하단 개방부를 구비한 상부 캡과, 상기 바닥벽 및 측벽을 구비하고 측벽에 의해 구획된 내측 공간으로 상기 상부 캡이 결합되는 하부 캡을 포함할 수 있고, 상기 상부 캡을 상기 하부 캡의 내측 공간 내로 압입할 때 상기 하부 캡의 바닥벽은 상기 상부 캡의 하단에 의해 가압되어 파열되는 파열선을 구비할 수 있다.In the gene amplification and detection device using isothermal amplification and lateral flow analysis of one embodiment of the present invention, the tube cap includes an upper cap having the upper wall, a side wall dividing the internal space, and a lower opening, and the bottom wall. and a lower cap that has a side wall and the upper cap is coupled to an inner space defined by the side wall, and when the upper cap is pressed into the inner space of the lower cap, the bottom wall of the lower cap is connected to the upper cap. It may have a rupture line that ruptures when pressed by the bottom of the cap.
본 발명의 일 구체예의 등온증폭 및 측방유동분석을 이용한 유전자 증폭 및 검출장치에 있어서, 상기 바닥벽의 파열선은 십자형으로 형성될 수 있다.In the gene amplification and detection device using isothermal amplification and lateral flow analysis of one embodiment of the present invention, the rupture line of the bottom wall may be formed in a cross shape.
본 발명의 일 구체예의 등온증폭 및 측방유동분석을 이용한 유전자 증폭 및 검출장치에 있어서, 상기 상부 캡의 측벽 높이는 상기 하부 캡의 측벽 높이 보다 높고, 상기 상부 캡의 측벽 하단이 상기 하부 캡의 바닥벽에 맞닿은 상태로 상기 상부 캡을 상기 하부 캡에 결합하였을 때 상기 상부 캡의 상단과 상기 하부 캡의 상단 사이에 간극을 형성한다. In the gene amplification and detection device using isothermal amplification and lateral flow analysis of one embodiment of the present invention, the side wall height of the upper cap is higher than the side wall height of the lower cap, and the lower end of the side wall of the upper cap is the bottom wall of the lower cap. When the upper cap is coupled to the lower cap in a state of contact, a gap is formed between the upper end of the upper cap and the upper end of the lower cap.
본 발명의 일 구체예의 등온증폭 및 측방유동분석을 이용한 유전자 증폭 및 검출장치에 있어서, 상기 등온증폭기는 외통과, 상기 외통의 내측에 구비되어 상기 반응액 튜브를 수용하는 등온 챔버와, 상기 등온 챔버에 수용된 반응액 튜브에 일정한 온도유지를 위해 열을 전달하는 전열판과, 상기 전열판의 온도를 제어할 수 있는 온도제어 PCB 기판을 구비할 수 있다.In the gene amplification and detection device using isothermal amplification and lateral flow analysis of one embodiment of the present invention, the isothermal amplifier includes an external cylinder, an isothermal chamber provided inside the external cylinder to accommodate the reaction liquid tube, and the isothermal chamber. It may be provided with a heat transfer plate that transfers heat to maintain a constant temperature in the reaction liquid tube contained in and a temperature control PCB board that can control the temperature of the heat transfer plate.
본 발명의 일 구체예의 등온증폭 및 측방유동분석을 이용한 유전자 증폭 및 검출장치에 있어서, 상기 상부 캡은 폴리에틸렌 재질로 만들어질 수 있고, 상기 하부 캡은 실리콘 고무로 만들어질 수 있다. In the gene amplification and detection device using isothermal amplification and lateral flow analysis of one embodiment of the present invention, the upper cap may be made of polyethylene material, and the lower cap may be made of silicone rubber.
본 발명의 일 구체예의 등온증폭 및 측방유동분석을 이용한 유전자 증폭 및 검출장치에 있어서, 상기 반응액은 수산화칼륨 용액 및 폴리에틸렌글리콜을 포함할 수 있고, 상기 유전자 증폭용 반응물질은 Bst DNA 중합효소, 완충용액, 프라이머들, dNTP 및 Mg2+를 포함할 수 있다. In the gene amplification and detection device using isothermal amplification and lateral flow analysis of one embodiment of the present invention, the reaction solution may include potassium hydroxide solution and polyethylene glycol, and the gene amplification reactants include Bst DNA polymerase, It may contain buffer solution, primers, dNTP and Mg 2+ .
상기한 본 발명에 의하면, 등온증폭방법 및 측방유동분석법의 조합을 사용하므로 신속하고 간편하게 유전자 검출결과를 얻을 수 있다. According to the present invention described above, gene detection results can be obtained quickly and easily by using a combination of the isothermal amplification method and the lateral flow analysis method.
본 발명에 따르면, 반응액 튜브와 튜브 캡을 결합한 상태에서 상기 튜브 캡을 눌러주면 상기 반응액 튜브 내의 유전자 시료가 용해된 반응액과 상기 튜브 캡의 유전자 증폭용 반응물질이 상기 반응액 튜브 내에서 간단히 혼합될 수 있다.According to the present invention, when the tube cap is pressed while the reaction solution tube and the tube cap are combined, the reaction solution in which the gene sample in the reaction solution tube is dissolved and the reaction material for gene amplification in the tube cap are stored in the reaction solution tube. Can be easily mixed.
또한, 본 발명에 따르면, 튜브 캡과 반응액 튜브의 조립체를 등온증폭기의 등온 챔버에 삽입하여 유전자 시료를 등온증폭한 후, 상기 튜브 캡의 절개부를 통해, 멤브레인이 노출된 측방유동분석기의 개구부를 상기 반응액 튜브의 내부 공간 내에 넣게 되면 상기 반응액 튜브 내의 등온증폭 산물이 멤브레인과 접촉하고 모세관 현상에 의해 멤브레인을 따라 유동하면서 검출 대상 유전자의 존재 여부를 시각적으로 나타낼 수 있다. 따라서, 본 발명은 피펫을 사용하지 않으면서 외부의 영향을 받지 않고 등온증폭 및 측방유동분석을 이용한 유전자 증폭 및 검출을 신속하고 간편하게 수행할 수 있다. In addition, according to the present invention, the assembly of the tube cap and the reaction tube is inserted into the isothermal chamber of the isothermal amplifier to isothermally amplify the genetic sample, and then the opening of the lateral flow analyzer where the membrane is exposed is inserted through the incision in the tube cap. When placed in the inner space of the reaction tube, the isothermal amplification product in the reaction tube comes into contact with the membrane and flows along the membrane by capillary action, thereby visually indicating the presence or absence of the gene to be detected. Therefore, the present invention can quickly and easily perform gene amplification and detection using isothermal amplification and lateral flow analysis without using a pipette and without external influence.
뿐만 아니라, 본 발명은 등온증폭 산물이 얻어진 등온증폭기에 측방유동분석기를 간단히 결합하여 검출 대상 유전자를 검출할 수 있으므로, 분석물질의 오염을 방지할 수 있어 신뢰성 높은 결과를 얻을 수 있다. In addition, the present invention can detect the target gene by simply combining a lateral flow analyzer with the isothermal amplifier from which the isothermal amplification product is obtained, thereby preventing contamination of the analyte and obtaining highly reliable results.
추가로, 본 발명은 유전자 증폭 및 검출이 완료된 후 반응액 튜브 및 튜브 캡의 조립체와 측방유동분석기를 결합된 상태로 등온증폭기로부터 손쉽게 분리가능하므로 사용 후 제거가 간편하고 등온증폭기의 오염을 차단할 수 있는 장점이 있다.Additionally, in the present invention, after gene amplification and detection are completed, the assembly of the reaction tube and tube cap and the lateral flow analyzer can be easily separated from the isothermal amplifier, making it easy to remove after use and preventing contamination of the isothermal amplifier. There is an advantage.
한편, 전술한 바와 같은 효과들에 의해 본 발명의 범위가 제한되는 것은 아니다. Meanwhile, the scope of the present invention is not limited by the effects described above.
도 1은 본 발명의 일 구체예에 따른 유전자 증폭 및 검출장치를 구성하는 부품들을 분리하여 도시한 분해사시도다.Figure 1 is an exploded perspective view showing the parts constituting the gene amplification and detection device according to an embodiment of the present invention separated.
도 2는 본 발명의 일 구체예에 따른 유전자 증폭 및 검출장치의 결합단면도이다. Figure 2 is a combined cross-sectional view of a gene amplification and detection device according to an embodiment of the present invention.
도 3a는 본 발명의 일 구체예에 따른 필름으로 밀봉된 반응액 튜브의 사시도이다.Figure 3a is a perspective view of a reaction liquid tube sealed with a film according to an embodiment of the present invention.
도 3b는 도 3a의 단면도이다.Figure 3b is a cross-sectional view of Figure 3a.
도 4a는 본 발명의 일 구체예에 따른 튜브 캡의 사시도이다.Figure 4A is a perspective view of a tube cap according to one embodiment of the present invention.
도 4b는 본 발명의 일 구체예에 따른 튜브 캡의 저면 사시도이다.4B is a bottom perspective view of a tube cap according to one embodiment of the present invention.
도 4c는 도 4a의 'I-I'선을 따라 절단하여 도시한 튜브 캡의 단면도이다.Figure 4c is a cross-sectional view of the tube cap taken along line 'I-I' of Figure 4a.
도 5a는 본 발명의 일 구체예에 따라 튜브 캡을 반응액 튜브에 결합한 상태의 단면도이다. Figure 5a is a cross-sectional view of a tube cap coupled to a reaction liquid tube according to an embodiment of the present invention.
도 5b는 도 5a의 상태에서 튜브 캡의 상부 캡을 가압하여 하부 캡 내로 압입한 상태의 결합단면도이다.Figure 5b is a cross-sectional view of the tube cap in the state of Figure 5a when the upper cap of the tube cap is pressed and pressed into the lower cap.
도 6a는 본 발명의 일 구체예에 따라 튜브 캡과 반응액 튜브의 조립체를 등온증폭기의 등온 챔버 내에 삽입하기 전(前) 상태의 결합단면도이다. Figure 6a is a cross-sectional view of the assembly of the tube cap and the reaction liquid tube before inserting it into the isothermal chamber of the isothermal amplifier according to one embodiment of the present invention.
도 6b는 본 발명의 일 구체예에 따라 튜브 캡과 반응액 튜브의 조립체를 등온증폭기의 등온 챔버 내에 삽입하여 등온증폭하는 상태의 결합단면도이다. Figure 6b is a cross-sectional view of the assembly of the tube cap and the reaction tube being inserted into the isothermal chamber of the isothermal amplifier and performing isothermal amplification according to an embodiment of the present invention.
도 7a는 본 발명의 일 구체예에 따른 측방유동분석기의 사시도이다. Figure 7a is a perspective view of a lateral flow analyzer according to an embodiment of the present invention.
도 7b는 본 발명의 일 구체예에 따른 측방유동분석기의 단면도이다. Figure 7b is a cross-sectional view of a lateral flow analyzer according to an embodiment of the present invention.
이하, 첨부된 도면을 참조하여 본 발명의 바람직한 실시예를 설명하면 다음과 같다. 본 발명의 하기 실시예는 본 발명을 구체화하기 위한 것일 뿐 본 발명의 권리범위를 제한하거나 한정하는 것이 아님은 물론이다. 본 발명의 상세한 설명 및 실시예로부터 본 발명이 속하는 기술분야의 전문가가 용이하게 유추할 수 있는 것은 본 발명의 권리범위에 속하는 것으로 해석된다. 본 발명에 인용된 참고문헌들은 본 발명에 참고로서 통합된다.Hereinafter, preferred embodiments of the present invention will be described with reference to the attached drawings. Of course, the following examples of the present invention are only intended to embody the present invention and do not limit or limit the scope of the present invention. Anything that can be easily inferred by an expert in the technical field to which the present invention belongs from the detailed description and examples of the present invention will be interpreted as falling within the scope of the rights of the present invention. References cited herein are hereby incorporated by reference.
명세서 전체에서, 어떤 부분이 어떤 구성 요소를 "포함"한다고 할 때, 이는 특별히 반대되는 기재가 없는 한 다른 구성 요소를 제외하는 것이 아니라 다른 구성 요소를 더 포함할 수 있는 것을 의미한다.Throughout the specification, when a part is said to “include” a certain component, this means that it may further include other components rather than excluding other components, unless specifically stated to the contrary.
도 1 및 도 2에 도시된 바와 같이, 본 발명의 일 구체예에 따른 등온증폭기와 측방유동분석기를 이용한 유전자 증폭 및 검출장치는, 상단에 개방부를 구비하고 내부공간에는 반응액(L)(유전자 시료가 혼합되어 용해됨)을 수용하는 반응액 튜브(10)와, 반응액 튜브(10)의 개방부에 결합되어 반응액 튜브(10)의 내부를 밀봉하는 튜브 캡(20)과, 튜브 캡(20)이 결합된 반응액 튜브(10)를 수용하고 등온 상태를 유지하여 반응액 튜브(10) 내의 유전자 시료를 등온증폭하는 등온증폭기(30)와, 튜브 캡(20)과 반응액 튜브(10)의 조립체에 삽입 결합되고 등온증폭기(30)에서 증폭된 반응액 튜브(10) 내의 등온증폭 산물을 멤브레인(47)과 접촉시켜 특정의 표적 유전자를 검출하는 측방유동분석기(40)를 포함한다. 이하 본 발명의 구성요소들 및 사용방법에 대하여 설명한다. As shown in Figures 1 and 2, the gene amplification and detection device using an isothermal amplifier and a lateral flow analyzer according to an embodiment of the present invention has an opening at the top and contains a reaction solution (L) (gene a reaction liquid tube 10 that accommodates the sample (the sample is mixed and dissolved), a tube cap 20 that is coupled to the opening of the reaction liquid tube 10 and seals the inside of the reaction liquid tube 10, and a tube cap. (20) accommodates the combined reaction solution tube 10 and maintains an isothermal state to isothermally amplify the gene sample in the reaction solution tube 10, an isothermal amplifier 30, a tube cap 20, and a reaction solution tube ( It includes a lateral flow analyzer (40) that is inserted into the assembly of 10) and detects a specific target gene by contacting the isothermal amplification product in the reaction tube (10) amplified in the isothermal amplifier (30) with the membrane (47). . Hereinafter, the components and method of use of the present invention will be described.
<반응액 튜브(10)><Reaction liquid tube (10)>
도 3a 및 도 3b에 도시된 바와 같이, 반응액 튜브(10)는 내부공간에 반응액(L)을 수용하고, 사용 전에는 상단의 개방부(11)에 부착된 필름(15)으로 밀봉되어 그 내부가 외부로부터 차단되어 있다. As shown in FIGS. 3A and 3B, the reaction liquid tube 10 accommodates the reaction liquid (L) in its internal space, and is sealed with a film 15 attached to the opening 11 at the top before use. The inside is blocked from the outside.
사용자는 면봉(미도시)을 콧속에 넣고 콧속을 문지르면 면봉에 검체가 채취되고, 반응액 튜브(10)의 필름(15)을 제거한 후 검체가 묻은 면봉을 반응액 튜브(10) 내의 반응액(L)에 넣으면 면봉에 묻은 검체 중 유전자 시료가 반응액(L) 내에 용해된다. 반응액 튜브(10) 내에 수용된 반응액(L)은 수산화칼륨 용액 및 폴리에틸렌글리콜을 포함하고, 면봉에 묻은 검체로부터 유전자 시료를 용해시킨다. 이와 같은 작업이 완료된 후, 반응액 튜브(10)의 개방부(11)에 튜브 캡(20)을 결합한다. 반응액 튜브(10)는 예를 들어 폴리프로필렌 재질로 만들어질 수 있으나, 이에 제한되는 것이 아니며, 안정적인 반응액 보관 및 유전자 증폭에 적합한 다양한 재질이 사용될 수 있음은 물론이다.The user inserts a cotton swab (not shown) into the nose and rubs the inside of the nose to collect a sample on the swab. After removing the film 15 of the reaction liquid tube 10, the cotton swab with the sample is placed in the reaction liquid tube 10 ( When placed in L), the genetic sample from the sample on the cotton swab is dissolved in the reaction solution (L). The reaction solution (L) contained in the reaction solution tube 10 contains potassium hydroxide solution and polyethylene glycol, and dissolves the genetic sample from the specimen affixed to the cotton swab. After this work is completed, the tube cap 20 is coupled to the opening 11 of the reaction liquid tube 10. The reaction solution tube 10 may be made of, for example, polypropylene, but is not limited thereto, and of course, various materials suitable for stable reaction solution storage and gene amplification may be used.
< 튜브 캡(20)><Tube cap (20)>
도 4a 내지 도 4c에 도시된 튜브 캡(20)은 상부 캡(21)과 하부 캡(26)으로 이루어진다. 상부 캡(21)은 상단 벽에 절개부(23)가 형성되고, 내부공간(S20)을 구획하는 측벽을 구비하며, 하단은 개방되어 개방부를 형성한다. 하부 캡(26)은 측벽에 의해 내측 공간이 형성되어 있고 상단은 개방되어 개방부를 형성한다. The tube cap 20 shown in FIGS. 4A to 4C consists of an upper cap 21 and a lower cap 26. The upper cap 21 has a cutout 23 formed on the upper wall, has a side wall that partitions the internal space S 20 , and the lower end is open to form an opening. The lower cap 26 has an inner space formed by the side wall and the upper end is open to form an opening.
상부 캡(21)의 측벽이 하부 캡(26)의 상단 개방부를 통해 하부 캡(26)의 내측 공간 내로 이동가능하게 끼워져 결합된다. 하부 캡(20)의 하단의 바닥벽(28)에는 '십자(+)'형 파열선(27)이 형성되어 있다. 상부 캡(21)의 내부공간(S20)에는 유전자 증폭용 반응물질(p)이 수용된다. 유전자 증폭용 반응물질(p)은 동결건조될 수 있으며, 동결건조된 유전자 증폭용 반응물질(p)은 유전자 시료의 등온증폭에 필요한 Bst DNA 중합효소, 완충용액, 프라이머들, dNTP 및 Mg2+를 포함한다. The side wall of the upper cap 21 is movably fitted and coupled into the inner space of the lower cap 26 through the upper opening of the lower cap 26. A 'cross (+)' shaped rupture line 27 is formed on the bottom wall 28 of the lower end of the lower cap 20. The reaction material (p) for gene amplification is accommodated in the internal space (S 20 ) of the upper cap (21). The reactant for gene amplification (p) can be freeze-dried, and the freeze-dried reactant for gene amplification (p) contains Bst DNA polymerase, buffer solution, primers, dNTP, and Mg 2+ required for isothermal amplification of gene samples. Includes.
상부 캡(21)의 측벽 하단이 하부 캡(26)의 측벽에 의해 형성된 내측 공간 내로 안내되어 끼워져 결합될 때, 하부 캡(26)의 바닥벽(28)이 상부 캡(21)의 내부공간(S20)의 하단 개방부를 폐쇄함으로써 내부공간(S20)에 유전자 증폭용 반응물질(p)을 안전하게 수용할 수 있게 된다. When the lower end of the side wall of the upper cap 21 is guided and fitted into the inner space formed by the side wall of the lower cap 26, the bottom wall 28 of the lower cap 26 is connected to the inner space of the upper cap 21 ( By closing the lower opening of S 20 ), the reaction material (p) for gene amplification can be safely accommodated in the internal space (S 20 ).
도 4c에 도시된 바와 같이, 상부 캡(21)의 측벽 높이(h1)는 하부 캡(26)의 측벽 높이(h2) 보다 높다. 따라서 상부 캡(21)의 측벽 하단이 하부 캡(26)의 바닥벽(28)에 맞닿은 상태로 상부 캡(21)을 하부 캡(26)에 결합하였을 때, 상부 캡(21)의 상단과 하부 캡(26)의 상단 사이에 간극(D)을 형성하게 된다. As shown in FIG. 4C, the side wall height h1 of the upper cap 21 is higher than the side wall height h2 of the lower cap 26. Therefore, when the upper cap 21 is coupled to the lower cap 26 with the lower end of the side wall of the upper cap 21 in contact with the bottom wall 28 of the lower cap 26, the upper and lower sides of the upper cap 21 A gap D is formed between the tops of the caps 26.
한편, 상부 캡(21)은 폴리에틸렌 재질로 만들어질 수 있고, 하부 캡(26)은 실리콘 고무로 만들어질 수 있다. 그러나, 상부 캡(21)과 하부 캡(26)은 상기한 재질에 제한되는 것이 아니며, 안정적인 유전자 증폭용 반응물질 보관, 반응용 튜브(10)와 결합 시 유전자 증폭용 반응물질의 효율적인 탈거 및 낙하, 그리고 유전자 증폭에 적합한 다양한 재질이 사용될 수 있음은 물론이다. Meanwhile, the upper cap 21 may be made of polyethylene material, and the lower cap 26 may be made of silicone rubber. However, the upper cap 21 and the lower cap 26 are not limited to the above-described materials, and provide stable storage of reactants for gene amplification and efficient removal and dropping of reactants for gene amplification when combined with the reaction tube 10. , and of course, various materials suitable for gene amplification can be used.
<반응액 튜브(10)에 튜브 캡(20)의 결합><Coupling of the tube cap (20) to the reaction liquid tube (10)>
도 3b, 도 4c 및 도 5a에 도시된 바와 같이, 상부 캡(21) 및 하부 캡(26)이 조립된 튜브 캡(20)은 비닐(15)이 제거된 반응액 튜브(10)의 개방부(11)에 결합된다 이에 따라, 유전자 시료가 용해된 반응액을 수용하는 반응액 튜브(10)의 내부공간을 밀폐하게 되고 내부공간 안으로 이물질이 침입하지 않게 된다. As shown in FIGS. 3B, 4C, and 5A, the tube cap 20 with the upper cap 21 and lower cap 26 assembled is the opening of the reaction liquid tube 10 from which the vinyl 15 has been removed. It is coupled to (11). Accordingly, the inner space of the reaction solution tube (10) containing the reaction solution in which the genetic sample is dissolved is sealed, and foreign substances are prevented from entering the internal space.
그런 다음, 도 5a의 화살표로 표시된 바와 같이, 튜브 캡(20)을 반응액 튜브(10)의 개방부(11)에 결합한 상태에서, 상부 캡(21)의 상단 벽을 눌러서 가압하면, 상부 캡(21)의 측벽 하단이 하부 캡(26)의 내측 공간 내로 간극(D) 만큼 진입한다. 이에 따라, 상부 캡(21)의 측벽 하단이 하부 캡(26)의 바닥벽(28)을 가압하게 되고, 파열선(27)은 가압된 외력에 의해 파열된다. 파열선(27)의 파열에 따라 바닥벽(28)은 4등분되면서 도 5a의 화살표 방향으로 회전함으로써 상부 캡(21)의 내부공간(S20)이 개봉된다. 파열선(27)의 파열에 따라 나누어진 바닥벽(28)이 도 5b에 도시되어 있다.Then, as indicated by the arrow in FIG. 5A, when the tube cap 20 is coupled to the opening 11 of the reaction liquid tube 10 and the upper wall of the upper cap 21 is pressed, the upper cap The lower end of the side wall of (21) enters the inner space of the lower cap (26) by the gap D. Accordingly, the lower end of the side wall of the upper cap 21 presses the bottom wall 28 of the lower cap 26, and the rupture line 27 is ruptured by the pressed external force. As the rupture line 27 ruptures, the bottom wall 28 is divided into four parts and rotates in the direction of the arrow in FIG. 5A, thereby opening the inner space S 20 of the upper cap 21. The bottom wall 28 divided according to the rupture of the rupture line 27 is shown in Figure 5b.
하부 캡(26) 및 바닥벽(28)은 탄성이 있는 실리콘 고무로 만들어지기 때문에, 바닥벽(28)은 상부 캡(21)의 측벽 하단에 의해 탄력적으로 눌려지면서 분단되고, 탄성을 갖는 분단된 바닥벽(28)은 반응액 튜브(10)의 측벽 내부와 상부 캡(21)의 측벽 사이에 밀착되어 끼워지게 된다.Since the lower cap 26 and the bottom wall 28 are made of elastic silicone rubber, the bottom wall 28 is divided while being elastically pressed by the lower side of the side wall of the upper cap 21, and the elastic divided The bottom wall 28 is tightly fitted between the inside of the side wall of the reaction liquid tube 10 and the side wall of the upper cap 21.
그리고 나서, 도 5b에 도시된 바와 같이, 상부 캡(21)의 내부공간(S20)에 수용된 유전자 증폭용 반응물질(p)은 튜브 캡(20)으로부터 반응액 튜브(10)로 낙하하여 반응액 튜브(10) 내의 반응액(L)에 용해되어 혼합된다. 따라서, 반응액 튜브(10)와 튜브 캡(20)을 결합한 상태에서 튜브 캡(20)을 눌러주면 반응액 튜브(10) 내의 유전자 시료가 용해된 반응액(L)과 튜브 캡(20)의 유전자 증폭용 반응물질(p)이 반응액 튜브(10) 내에서 간단히 혼합될 수 있다.Then, as shown in Figure 5b, the reaction material (p) for gene amplification accommodated in the inner space (S 20 ) of the upper cap (21) falls from the tube cap (20) to the reaction solution tube (10) and reacts. It is dissolved and mixed in the reaction liquid (L) in the liquid tube (10). Therefore, when the tube cap 20 is pressed while the reaction solution tube 10 and the tube cap 20 are combined, the genetic sample in the reaction solution tube 10 is dissolved in the reaction solution L and the tube cap 20. Reactants (p) for gene amplification can be simply mixed in the reaction tube (10).
한편, 유전자 시료가 용해되어 있는 반응액(L)과 유전자 증폭용 반응물질(p)(Bst DNA 중합효소, 완충용액, 프라이머들, dNTP 및 Mg2+ 포함)의 혼합액을 일정한 온도, 예를 들어 60℃ 내지 65℃에서 대략 30분 정도 반응시키면 검출대상이 되는 유전자가 등온증폭된다.Meanwhile, the mixture of the reaction solution (L) in which the genetic sample is dissolved and the reaction material for gene amplification (p) (including Bst DNA polymerase, buffer solution, primers, dNTP, and Mg 2+ ) is kept at a constant temperature, e.g. When reacted at 60°C to 65°C for approximately 30 minutes, the gene to be detected is isothermally amplified.
< 등온증폭기(30)><Isothermal amplifier (30)>
도 6a에 도시된 바와 같이, 등온증폭기(30)는 외통(31)과, 외통(31)의 내측에 구비되어 반응용 튜브(10)를 수용하는 등온 챔버(32)와, 등온 챔버(32)의 측벽 및 바닥 중 적어도 한 곳에 구비되어 등온 챔버(32)에 수용된 반응액 튜브(10)에 일정한 온도유지를 위해 열을 전달하는 전열판(34)과, 등온 챔버(32)의 바닥에 설치되어 전열판(34)의 온도를 제어할 수 있는 온도제어 PCB 기판(35)을 포함한다. As shown in FIG. 6A, the isothermal amplifier 30 includes an external cylinder 31, an isothermal chamber 32 provided inside the external cylinder 31 to accommodate the reaction tube 10, and an isothermal chamber 32. A heat transfer plate 34 is provided on at least one of the side walls and the bottom of the isothermal chamber 32 to transfer heat to the reaction liquid tube 10 accommodated in the isothermal chamber 32 to maintain a constant temperature, and a heat transfer plate is installed on the bottom of the isothermal chamber 32. It includes a temperature control PCB board (35) capable of controlling the temperature (34).
<등온증폭기를 이용한 등온증폭><Isothermal amplification using an isothermal amplifier>
도 6a 및 도 6b에 도시된 바와 같이, 튜브 캡(20)이 결합된 반응액 튜브(10)를 등온증폭기(30)의 등온 챔버(32) 내에 삽입하고, 온도제어 PCB 기판(35)을 가동시켜 전열판(34)을 가열하여 등온증폭기(30)의 등온 챔버(32)를 일정한 온도, 예를 들어 60℃ 내지 65℃에서 대략 30분 정도 유지하면, 반응액 튜브(10) 내의 반응액(L)에 용해된 유전자 시료가 유전자 증폭용 반응물질(p)(Bst DNA 중합효소, 완충용액, 프라이머들, dNTP 및 Mg2+ 포함)과 반응하여 검출대상이 되는 유전자가 등온증폭된다. As shown in FIGS. 6A and 6B, the reaction liquid tube 10 coupled with the tube cap 20 is inserted into the isothermal chamber 32 of the isothermal amplifier 30, and the temperature control PCB board 35 is operated. When the heating plate 34 is heated and the isothermal chamber 32 of the isothermal amplifier 30 is maintained at a constant temperature, for example, 60° C. to 65° C. for approximately 30 minutes, the reaction liquid (L) in the reaction liquid tube 10 ) reacts with the gene amplification reactant (p) (including Bst DNA polymerase, buffer solution, primers, dNTP, and Mg 2+ ), and the gene to be detected is isothermally amplified.
< 측방유동분석기(40)><Lateral flow analyzer (40)>
도 7a 및 도 7b에 도시된 바와 같이, 측방유동분석기(40)는 케이싱(41)과, 케이싱(41)의 하부에 형성되고, 반응액 튜브(10)와 튜브 캡(20)의 조립체와 결합할 때, 튜브 캡(20)의 절개부(23)와 4등분된 바닥벽(28) 사이의 공간을 관통하여 반응액 튜브(10)의 내부 공간으로 삽입되며, 하단면에는 개구부(44)를 가진 하부 첨단부(43)와, 케이싱(41) 내부에 길이방향을 따라 제공되고, 하단부는 하부 첨단부(43)의 개구부(44)를 통하여 반응액 튜브(10)의 내부 공간에 노출되어 반응액 튜브(10) 내의 등온증폭된 유전자 시료(액체 시료)와 접촉하게 되는 멤브레인(47)을 구비한다.As shown in FIGS. 7A and 7B, the lateral flow analyzer 40 is formed in the casing 41 and the lower part of the casing 41, and is coupled to the assembly of the reaction liquid tube 10 and the tube cap 20. When doing so, it penetrates the space between the cutout part 23 of the tube cap 20 and the quartered bottom wall 28 and is inserted into the inner space of the reaction liquid tube 10, and has an opening 44 on the bottom surface. A vibrating lower tip 43 and the casing 41 are provided along the longitudinal direction, and the lower end is exposed to the inner space of the reaction liquid tube 10 through the opening 44 of the lower tip 43 to react. It is provided with a membrane 47 that comes into contact with the isothermally amplified gene sample (liquid sample) in the liquid tube 10.
케이싱(41)은 좌측 케이싱부(41a)와 우측 케이싱부(41b)로 이루어지고, 그 내부에 길이방향으로 멤브레인(47)이 제공된다. The casing 41 consists of a left casing part 41a and a right casing part 41b, and a membrane 47 is provided therein in the longitudinal direction.
멤브레인(47)은 등온증폭된 유전자 시료(액체 시료)가 모세관 현상으로 흐를 수 있는 다공성 막으로 구성된다. 멤브레인(47)의 하류측에는 모세관 현상을 촉진하는 흡수패드(112)를 구비하고, 상류측에는 등온증폭된 유전자 시료(액체 시료)가 로딩되는 샘플패드(113)와 스트렙타비딘 또는 이와 동등한 물질이 접합된 금 나노입자들이 침지된 접합 패드(114)를 가진다. The membrane 47 is composed of a porous membrane through which the isothermally amplified gene sample (liquid sample) can flow through capillary action. On the downstream side of the membrane 47, there is an absorption pad 112 that promotes capillary action, and on the upstream side, a sample pad 113 on which isothermally amplified gene sample (liquid sample) is loaded and streptavidin or an equivalent material are conjugated. It has a bonding pad 114 in which gold nanoparticles are immersed.
< 측방유동분석기(40)의 결합>< Combination of lateral flow analyzer (40)>
등온증폭기(30)의 등온 챔버(32) 내에 수용된 반응액 튜브(10)와 튜브 캡(20)의 조립체의 상부 캡(21)의 상단 벽에 구비된 절개부(23)에 측방유동분석기(40)의 하부 첨단부(43)를 갖다 대고 눌러주면, 측방유동분석기(40)의 하부 첨단부(43)는 상부 캡(21)의 절개부(23)를 뚫고 들어가고 상부 캡(21)과 하부 캡(26)을 차례로 통과하여 반응액 튜브(10) 내의 등온증폭된 유전자 시료(액체 시료)에 잠기도록 반응액 튜브(10) 내로 진입한다(도 2 참조). A lateral flow analyzer (40) is inserted into the incision (23) provided on the upper wall of the upper cap (21) of the assembly of the reaction liquid tube (10) and the tube cap (20) accommodated in the isothermal chamber (32) of the isothermal amplifier (30). ), the lower tip 43 of the lateral flow analyzer 40 penetrates the cutout 23 of the upper cap 21 and separates the upper cap 21 and the lower cap. It sequentially passes through (26) and enters the reaction solution tube 10 to be immersed in the isothermally amplified gene sample (liquid sample) in the reaction solution tube 10 (see FIG. 2).
이 상태에서, 반응액 튜브(10)의 등온증폭된 유전자 시료(액체 시료)는 측방유동분석기(40)의 하부 첨단부(43)의 개구부(44)를 통하여 멤브레인(47) 표면과 접촉하게 되고, 이후 모세관 현상에 의해 등온증폭된 유전자 시료(액체 시료)는 멤브레인(47)을 따라 원활하게 유동된다. 유동되는 액체 시료 내에 검출대상이 되는 유전자가 존재하면 멤브레인(47)의 검출영역에서 밴드가 나타나게 된다. 이로써 검출대상이 되는 유전자, 예를 들어 특정 바이러스 유전자가 콧속에서 채취한 검체 내에 존재하는지 여부를 시각적으로 보여주게 되고, 사용자나 의료진은 특정 바이러스에 감염되었는지 여부를 용이하게 확인할 수 있다. 따라서, 본 발명에 따르면 피펫을 사용하지 않으면서 외부의 영향을 받지 않고 등온증폭 및 측방유동분석을 이용한 유전자 증폭 및 검출을 신속하고 간편하게 수행할 수 있다. In this state, the isothermally amplified gene sample (liquid sample) in the reaction tube 10 comes into contact with the surface of the membrane 47 through the opening 44 of the lower tip 43 of the lateral flow analyzer 40. , Afterwards, the gene sample (liquid sample) amplified isothermally by capillary action flows smoothly along the membrane 47. If a gene to be detected is present in the flowing liquid sample, a band appears in the detection area of the membrane 47. This visually shows whether a gene to be detected, for example, a specific viral gene, is present in a sample collected from the nose, and users or medical staff can easily check whether they are infected with a specific virus. Therefore, according to the present invention, gene amplification and detection using isothermal amplification and lateral flow analysis can be performed quickly and easily without using a pipette and without external influence.
한편, 측방유동분석기(40)에서 특정 표적 유전자의 존재 여부를 확인한 후, 반응액 튜브(10)와 튜브 캡(20)의 조립체에 결합된 측방유동분석기(40)를 등온증폭기(30)에서 분리한다. 측방유동분석기(40)는 반응액 튜브(10)와 튜브 캡(20)의 조립체와 결합한 상태로 등온증폭기(30)에서 분리되므로, 검사가 완료된 후 분석기구의 분리 및 폐기가 편리하다. 따라서, 본 발명은 사용 후 제거가 간편하고 등온증폭기의 오염을 차단할 수 있는 장점이 있다.Meanwhile, after confirming the presence of a specific target gene in the lateral flow analyzer 40, the lateral flow analyzer 40 coupled to the assembly of the reaction tube 10 and the tube cap 20 is separated from the isothermal amplifier 30. do. Since the lateral flow analyzer 40 is separated from the isothermal amplifier 30 while being coupled to the assembly of the reaction tube 10 and the tube cap 20, it is convenient to separate and dispose of the analysis device after the test is completed. Therefore, the present invention has the advantage of being easy to remove after use and preventing contamination of the isothermal amplifier.
이상, 본 발명을 상기 실시예를 들어 설명하였으나, 본 발명은 이에 제한되는 것이 아니다. 당업자라면 본 발명의 취지 및 범위를 벗어나지 않고 수정, 변경을 할 수 있으며 이러한 수정과 변경 또한 본 발명에 속하는 것임을 알 수 있을 것이다. Although the present invention has been described with reference to the above examples, the present invention is not limited thereto. Those skilled in the art will recognize that modifications and changes can be made without departing from the spirit and scope of the present invention, and that such modifications and changes also fall within the scope of the present invention.

Claims (7)

  1. 상단에 개방부를 구비하고, 내부공간에는 반응액이 수용되어 있으며, 유전자 시료가 상기 반응액에 혼합되어 용해되는 반응액 튜브와,A reaction solution tube having an opening at the top, containing a reaction solution in the inner space, and in which a genetic sample is mixed and dissolved in the reaction solution;
    상기 반응액 튜브의 개방부를 밀폐하도록 상기 개방부에 결합되고, 내부 공간에는 유전자 증폭용 반응물질이 수용되어 있으며, 상기 내부 공간의 상측을 구획하는 상단 벽에는 절개부를 구비하고, 상기 내부 공간의 하측을 구획하는 바닥벽에는 외력에 의해 파열되는 파열선을 구비하며, 상기 파열선이 외력에 의해 파열될 때 상기 유전자 증폭용 반응물질이 상기 반응액 튜브의 내부공간에 수용된 반응액으로 배출되어 혼합되는 튜브 캡과,It is coupled to the opening to seal the opening of the reaction solution tube, a reaction material for gene amplification is accommodated in the inner space, an upper wall dividing the upper side of the inner space is provided with a cutout, and a lower side of the inner space is provided. The bottom wall dividing the rupture line is provided with a rupture line that is ruptured by an external force, and when the rupture line is ruptured by an external force, the gene amplification reactant is discharged and mixed into the reaction solution contained in the inner space of the reaction solution tube. tube cap,
    상기 튜브 캡이 결합된 반응액 튜브를 수용하고, 상기 수용된 반응액 튜브를 일정한 온도에서 유지하여 상기 반응액 튜브의 내부공간 내의 상기 유전자 시료, 상기 반응액 및 상기 유전자 증폭용 반응물질의 혼합액을 반응시켜 상기 유전자 시료를 등온증폭시키는 등온증폭기와,The reaction solution tube to which the tube cap is coupled is accommodated, and the received reaction solution tube is maintained at a constant temperature to react the mixture of the genetic sample, the reaction solution, and the gene amplification reactant in the inner space of the reaction solution tube. an isothermal amplifier for isothermal amplification of the genetic sample,
    케이싱; 상기 케이싱의 하부에 형성되고, 하단에는 개방된 개구부를 가지며, 상기 튜브 캡의 절개부와 바닥벽을 관통하여 상기 반응액 튜브의 내부공간으로 삽입되는 하부 첨단부; 및 상기 케이싱 내부에 길이방향을 따라 제공되고, 상기 하부 첨단부의 개구부를 통하여 상기 반응액 튜브의 내부공간에 노출되어 상기 반응액 튜브 내의 등온증폭된 유전자 시료와 접촉하게 되는 멤브레인을 구비하는 측방유동분석기를 포함하는, 등온증폭 및 측방유동분석을 이용한 유전자 증폭 및 검출장치.casing; a lower tip formed at the lower part of the casing, having an open opening at the lower end, and inserted into the inner space of the reaction liquid tube through the cutout and bottom wall of the tube cap; and a lateral flow analyzer including a membrane provided along the longitudinal direction inside the casing and exposed to the inner space of the reaction solution tube through an opening at the lower tip and coming into contact with the isothermally amplified genetic sample within the reaction solution tube. Gene amplification and detection device using isothermal amplification and lateral flow analysis, including.
  2. 제1항에 있어서,According to paragraph 1,
    상기 튜브 캡은 상기 상단 벽, 상기 내부공간을 구획하는 측벽 및 하단 개방부를 구비한 상부 캡과, 상기 바닥벽 및 측벽을 구비하고 측벽에 의해 구획된 내측 공간으로 상기 상부 캡이 결합되는 하부 캡을 포함하고, The tube cap includes an upper cap having the top wall, a side wall defining the internal space, and a bottom opening, and a lower cap having the bottom wall and the side wall and the upper cap being coupled to the inner space defined by the side wall. Contains,
    상기 상부 캡을 상기 하부 캡의 내측 공간 내로 압입할 때 상기 하부 캡의 바닥벽은 상기 상부 캡의 하단에 의해 가압되어 파열되는 파열선을 구비하는, 등온증폭 및 측방유동분석을 이용한 유전자 증폭 및 검출장치. Gene amplification and detection using isothermal amplification and lateral flow analysis, wherein the bottom wall of the lower cap has a rupture line that is ruptured by being pressed by the bottom of the upper cap when the upper cap is pressed into the inner space of the lower cap. Device.
  3. 제2항에 있어서,According to paragraph 2,
    상기 바닥벽의 파열선은 십자형으로 형성되는, 등온증폭 및 측방유동분석을 이용한 유전자 증폭 및 검출장치.A gene amplification and detection device using isothermal amplification and lateral flow analysis, wherein the rupture line of the bottom wall is formed in a cross shape.
  4. 제2항 또는 제3항에 있어서,According to paragraph 2 or 3,
    상기 상부 캡의 측벽 높이는 상기 하부 캡의 측벽 높이 보다 높고, 상기 상부 캡의 측벽 하단이 상기 하부 캡의 바닥벽에 맞닿은 상태로 상기 상부 캡을 상기 하부 캡에 결합하였을 때 상기 상부 캡의 상단과 상기 하부 캡의 상단 사이에 간극을 형성하는, 등온증폭 및 측방유동분석을 이용한 유전자 증폭 및 검출장치.The side wall height of the upper cap is higher than the side wall height of the lower cap, and when the upper cap is coupled to the lower cap with the lower end of the side wall of the upper cap in contact with the bottom wall of the lower cap, the upper end of the upper cap and the A gene amplification and detection device using isothermal amplification and lateral flow analysis that forms a gap between the top of the lower cap.
  5. 제1항 내지 제3항 중 어느 한 항에 있어서,According to any one of claims 1 to 3,
    상기 등온증폭기는 외통과, 상기 외통의 내측에 구비되어 상기 반응액 튜브를 수용하는 등온 챔버와, 상기 등온 챔버에 수용된 반응액 튜브에 일정한 온도유지를 위해 열을 전달하는 전열판과, 상기 전열판의 온도를 제어할 수 있는 온도제어 PCB 기판을 구비하는, 등온증폭 및 측방유동분석을 이용한 유전자 증폭 및 검출장치.The isothermal amplifier includes an external cylinder, an isothermal chamber provided inside the external cylinder to accommodate the reaction liquid tube, a heat transfer plate that transfers heat to the reaction liquid tube accommodated in the isothermal chamber to maintain a constant temperature, and a temperature of the heat transfer plate. Gene amplification and detection device using isothermal amplification and lateral flow analysis, including a temperature control PCB board capable of controlling.
  6. 제2항 또는 제3항에 있어서,According to paragraph 2 or 3,
    상기 상부 캡은 폴리에틸렌 재질로 만들어지고, 상기 하부 캡은 실리콘 고무로 만들어지는, 등온증폭 및 측방유동분석을 이용한 유전자 증폭 및 검출장치.Gene amplification and detection device using isothermal amplification and lateral flow analysis, wherein the upper cap is made of polyethylene material and the lower cap is made of silicone rubber.
  7. 제1항 내지 제3항 중 어느 한 항에 있어서,According to any one of claims 1 to 3,
    상기 반응액은 수산화칼륨 용액 및 폴리에틸렌글리콜을 포함하고, 상기 유전자 증폭용 반응물질은 Bst DNA 중합효소, 완충용액, 프라이머들, dNTP 및 Mg2+를 포함하는, 등온증폭 및 측방유동분석을 이용한 유전자 증폭 및 검출장치.The reaction solution contains potassium hydroxide solution and polyethylene glycol, and the reaction materials for gene amplification include Bst DNA polymerase, buffer solution, primers, dNTP, and Mg 2+ , and gene amplification using isothermal amplification and lateral flow analysis. Amplification and detection device.
PCT/KR2022/020560 2022-03-07 2022-12-16 Gene amplification and detection device using isothermal amplification and lateral flow assay WO2023171888A1 (en)

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KR20140142161A (en) * 2013-05-30 2014-12-11 케이맥(주) Real time quantitative and qualitative analizing method
KR101998842B1 (en) * 2018-08-17 2019-07-11 웰스바이오 주식회사 Tube assembly for dripping to diagnostic strip and extracting gene fragment
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WO2021263003A1 (en) * 2020-06-24 2021-12-30 President And Fellows Of Harvard College Automatic multi-step reaction device

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KR101157042B1 (en) 2005-04-18 2012-06-21 주식회사 엘지생명과학 Improved Lateral Flow Immunoassay And Device Therefor
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KR101998842B1 (en) * 2018-08-17 2019-07-11 웰스바이오 주식회사 Tube assembly for dripping to diagnostic strip and extracting gene fragment
KR102069602B1 (en) * 2018-10-15 2020-01-23 주식회사 알엠생명과학 Biopsy Container
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