WO2023080305A1

WO2023080305A1 - Integrated molecular diagnostic device and molecular diagnostic method using same

Info

Publication number: WO2023080305A1
Application number: PCT/KR2021/016371
Authority: WO
Inventors: 이국녕; 이현영; 조상규; 홍동기; 천정훈; 강주성; 강혜림; 윤서진
Original assignee: 주식회사 위즈바이오솔루션
Priority date: 2021-11-03
Filing date: 2021-11-10
Publication date: 2023-05-11

Abstract

The present invention relates to an integrated molecular diagnostic device and a molecular diagnostic method using same, the device comprising: a buffer tube into which a sample collection tool for collecting a sample is inserted and in which a sample solution that contains a nucleic acid extracted from the collected sample is generated; a cartridge which is coupled to the buffer tube and receives the sample solution, conveys the sample solution to a reaction chamber via a fluid channel, and is supplied with heat of a certain temperature to perform a nucleic acid amplification reaction; and a diagnostic module body which is detachably coupled to the cartridge and supplies the reaction chamber with the heat of a certain temperature, and detects the nucleic acid amplification reaction to diagnose whether a diagnosis target is present.

Description

All-in-one molecular diagnostic device and molecular diagnostic method using the same

The present invention relates to an all-in-one molecular diagnosis device and a molecular diagnosis method using the same, and more particularly, can be performed with a single device with minimal user intervention from pretreatment of a collected sample to molecular diagnosis, and can be manufactured in a small size. It relates to an integrated molecular diagnostic device capable of on-site diagnosis and a molecular diagnostic method using the same.

The national research and development projects supporting the present invention are as follows.

Assignment identification number 1465032760

Assignment No. 4W400820

Ministry Name Ministry of Health and Welfare

Task management (specialized) agency name Korea Health Industry Development Institute

Research project name Infectious disease prevention technology development project

Development of a molecular diagnostic device with integrated LAMP method for sample preprocessing capable of rapid on-site diagnosis

Contribution rate

1/1

Name of the project performing organization Wiz Bio Solution Co., Ltd.

Research Period 2020.09.01 ~ 2023.02.28

In general, molecular diagnosis methods are more accurate than immunodiagnosis methods because they directly test the genes of harmful bacteria or viruses, and can more accurately diagnose the causative bacteria of infection. However, the molecular diagnosis method is relatively complicated in its procedure because sample collection, cell disruption, nucleic acid extraction, and nucleic acid amplification processes are sequentially performed, and it takes about 30 minutes to 2 hours to obtain results.

Therefore, research is being conducted to shorten the test time of molecular diagnostic methods and quickly perform pre-processing of samples in order to apply them to point of care testing (POCT). In general, sample pretreatment is to extract nucleic acids (DNA, RNA, etc.) from cells to be amplified through a polymerase chain reaction (PCR). Components that hinder or inhibit the amplification reaction are removed and only target nucleic acids are purified. to be highly refined.

A traditional sample preparation method is a method of extracting nucleic acids using a centrifugal separator, but recently, a technique for automating the sample preparation process without using a centrifugal separator has been developed. Accordingly, a cartridge-integrated molecular diagnosis device that performs sample preprocessing and molecular diagnosis with one diagnosis device is being commercialized.

In general, cartridge-integrated molecular diagnostic devices mechanically control fluid by using valves or motors to transport samples from sample pretreatment to amplification. Therefore, the cartridge structure and control method are complicated. In addition to these methods, there is a method of controlling a small volume of fluid using an electrowetting method, but since the manufacture of an electrode array is essential, the cartridge cost is expensive and the reliability of the diagnosis result is relatively low.

One embodiment of the present invention is an all-in-one molecular diagnosis device that can perform on-site diagnosis by minimizing user intervention from pretreatment of a collected sample to molecular diagnosis, and can be manufactured in a small size, and a molecular diagnosis method using the same want to provide

In embodiments, an integrated molecular diagnostic device may include: a buffer tube into which a sampling tool for collecting a sample is inserted and generating a sample solution containing nucleic acid extracted from the collected sample; a cartridge coupled to the buffer tube to receive the sample solution, and carrying out a nucleic acid amplification reaction by transferring the sample solution to a reaction chamber through a fluidic channel; and a diagnostic module main body detachably coupled to the cartridge to supply heat of a certain temperature to the reaction chamber and detecting the nucleic acid amplification reaction to diagnose the presence or absence of a diagnostic target.

Among the embodiments, a molecular diagnosis method using an integrated molecular diagnosis device includes collecting a sample using a sampling tool; preparing a sample solution by extracting nucleic acids from the sample while the buffer tube is mounted on the cartridge and the cartridge is inserted into the main body of the diagnostic module; discharging the sample solution from the buffer tube and transferring the sample solution to a reaction chamber through a fluid channel formed in the cartridge when the cartridge is inserted into the diagnostic module body with the buffer tube inserted; performing a nucleic acid amplification reaction by applying heat of a certain temperature to the reaction chamber; and detecting the nucleic acid amplification reaction to diagnose the presence or absence of a diagnosis target.

The disclosed technology may have the following effects. However, it does not mean that a specific embodiment must include all of the following effects or only the following effects, so it should not be understood that the scope of rights of the disclosed technology is limited thereby.

The all-in-one molecular diagnosis device and molecular diagnosis method using the same according to an embodiment of the present invention can be performed with a single device by minimizing user intervention from pre-processing of collected samples to molecular diagnosis, and can be manufactured in a small size for on-site diagnosis. this is possible

1 is a block diagram showing a molecular diagnostic system according to an embodiment of the present invention.

FIG. 2 is a diagram illustrating an embodiment of the integrated molecular diagnostic device shown in FIG. 1 .

FIG. 3 is a view for explaining the buffer tube shown in FIG. 1;

4a to 4c are views for explaining the opening and closing body shown in FIG. 3 .

5 is a diagram for explaining the cartridge shown in FIG. 1;

6a to 6c are views for explaining the cartridge body shown in FIG.

7A and 7B are diagrams for explaining the cartridge holder shown in FIG. 5 .

FIG. 8 is a block diagram for explaining the diagnosis module main body shown in FIG. 1 .

FIG. 9 is a view for explaining the body shown in FIG. 8;

FIG. 10 is a diagram for explaining the heat supply module shown in FIG. 8 .

FIG. 11 is a diagram for explaining the detection module shown in FIG. 8 .

12 is a flowchart illustrating a molecular diagnosis method according to an embodiment of the present invention.

13 is a view for explaining a movement operation of the inlet stopper of the buffer tube.

14 is a diagram for explaining a movement path of a sample solution.

15A and 15B are diagrams for explaining diagnosis results.

FIG. 16 is a view showing another embodiment of the integrated molecular diagnostic device shown in FIG. 1 .

17A to 17C are diagrams for explaining the cartridge shown in FIG. 16;

FIG. 18 is a diagram for explaining the diagnosis module main body shown in FIG. 16 .

FIG. 19 is a diagram for explaining the heat supply module shown in FIG. 18 .

20A and 20B are diagrams for explaining the detection module shown in FIG. 18 .

FIG. 21 is a diagram for explaining a detection signal output from the detection module shown in FIG. 18 .

Since the description of the present invention is only an embodiment for structural or functional description, the scope of the present invention should not be construed as being limited by the embodiments described in the text. That is, since the embodiment can be changed in various ways and can have various forms, it should be understood that the scope of the present invention includes equivalents capable of realizing the technical idea. In addition, since the object or effect presented in the present invention does not mean that a specific embodiment should include all of them or only such effects, the scope of the present invention should not be construed as being limited thereto.

Meanwhile, the meaning of terms described in this application should be understood as follows.

Terms such as "first" and "second" are used to distinguish one component from another, and the scope of rights should not be limited by these terms. For example, a first element may be termed a second element, and similarly, a second element may be termed a first element.

It should be understood that when an element is referred to as being “connected” to another element, it may be directly connected to the other element, but other elements may exist in the middle. On the other hand, when an element is referred to as being "directly connected" to another element, it should be understood that no intervening elements exist. Meanwhile, other expressions describing the relationship between components, such as “between” and “immediately between” or “adjacent to” and “directly adjacent to” should be interpreted similarly.

Expressions in the singular number should be understood to include plural expressions unless the context clearly dictates otherwise, and terms such as “comprise” or “having” refer to an embodied feature, number, step, operation, component, part, or these. It should be understood that it is intended to indicate that a combination exists, and does not preclude the possibility of the presence or addition of one or more other features, numbers, steps, operations, components, parts, or combinations thereof.

In each step, the identification code (eg, a, b, c, etc.) is used for convenience of explanation, and the identification code does not describe the order of each step, and each step clearly follows a specific order in context. Unless otherwise specified, it may occur in a different order than specified. That is, each step may occur in the same order as specified, may be performed substantially simultaneously, or may be performed in the reverse order.

The present invention can be implemented as computer readable code on a computer readable recording medium, and the computer readable recording medium includes all types of recording devices in which data readable by a computer system is stored. . Examples of computer-readable recording media include ROM, RAM, CD-ROM, magnetic tape, floppy disk, and optical data storage devices. In addition, the computer-readable recording medium may be distributed to computer systems connected through a network, so that computer-readable codes may be stored and executed in a distributed manner.

All terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which the present invention belongs, unless defined otherwise. Terms defined in commonly used dictionaries should be interpreted as consistent with meanings in the context of the related art, and cannot be interpreted as having ideal or excessively formal meanings unless explicitly defined in the present application.

FIG. 1 is a block diagram showing a molecular diagnosis system according to an embodiment of the present invention, and FIG. 2 is a diagram showing an embodiment of the integrated molecular diagnosis device shown in FIG. 1 .

Referring to FIGS. 1 and 2 , an integrated molecular diagnosis system according to an embodiment of the present invention may include an integrated molecular diagnosis device 1 and a user terminal 2 . The integrated molecular diagnostic device 1 may communicate with the user terminal 2 through a network. Here, the network may include various types of mobile communication networks such as wired communication networks, wireless LANs, Wi-Fi, Bluetooth, wireless communication networks such as ZigBee, 2G, 3G, 4G, 5G, and LTE.

The integrated molecular diagnosis device 1 can automatically pre-process a sample collected and injected from a user to be tested, and diagnose the presence or absence of a diagnosis target in real time from the pre-processed sample. A diagnosis subject according to an embodiment of the present invention may be a causative agent of a respiratory disease. For example, the diagnosis target may be a respiratory disease causative bacterium molecule such as RSV (respiratory syncytial virus), COVID-19, delta COVID-19, and the like. The integrated molecular diagnosis device 1 may transmit diagnosis results to the user terminal 2 .

The integrated molecular diagnostic device 1 may include a sampling tool 100 , a buffer tube 200 , a cartridge 300 and a diagnostic module body 400 . Here, the sampling tool 100, the buffer tube 200, and the cartridge 300 are disposable and may be discarded after use. The sampling tool 100 collects a sample from a user to be tested. The sampling tool 100 may collect a sample from the mucous membrane of the inner wall of the nasal cavity or oral cavity of a user to be examined in order to diagnose a respiratory disease causative bacteria. The sampling tool 100 may be formed of a shape and material that can easily collect a sample from a user to be tested, and may be formed, for example, in the form of a cotton swab that can be inserted into the nasal cavity or oral cavity of the user to be tested. can

The buffer tube 200 contains a pre-injected buffer solution, and accommodates the sampling tool 100 to be immersed in the buffer solution. Here, the buffer solution is a lysis buffer, which is a buffer solution used when destroying cell membranes, and microparticles for improving lysis efficiency may be mixed and injected into the buffer tube 200 in advance.

The buffer tube 200 extracts nucleic acids from the sample collected through the sampling tool 100 to create a sample solution. In general, the method of destroying the cell membrane of the sample is a chemical method of adjusting the pH of the buffer solution, a method of removing large protein molecules through protein denaturation by heating the buffer solution to a certain temperature (60 ~ 95 ℃), or a method of ultrasonic wave There are methods such as applying physical shock using it.

Since the respiratory causative bacteria sample has relatively few impurities compared to blood or other samples, one embodiment of the present invention applies a method of destroying cell membranes by applying physical shock to the sample by shaking the buffer tube 200. That is, the sample collection tool 100 is inserted into the buffer tube 200 and shaken together with the buffer solution in a sealed state, and nucleic acids can be extracted by destroying cell membranes of the sample by the shaking operation. An embodiment of the present invention is not limited thereto, and nucleic acids may be extracted by using at least one of the methods of heating a buffer solution or applying physical impact using ultrasonic waves.

The buffer tube 200 may be inserted into the cartridge 300 and supply a sample solution from which nucleic acids are extracted to the cartridge 300 . When the buffer tube 200 is inserted into the cartridge 300, the sample solution may be discharged to the outside by a hole in the bottom surface. To this end, the buffer tube 200 may be formed of a plastic material that has excellent chemical resistance and is not hard. For example, the buffer tube 200 may be formed of polypropylene (PP) or polycarbonate (PC).

The cartridge 300 is coupled to the buffer tube 200 to receive a sample solution from the buffer tube 200 . The cartridge 300 extracts a fixed amount of sample solution through at least one fluid channel, mixes it with a pre-injected reagent, and receives heat of a constant temperature from the diagnostic module body 400 to perform a nucleic acid amplification reaction.

Here, the reagent is for amplifying nucleic acid included in the sample solution to detect a diagnosis target, and may be pre-injected into the cartridge 300 in a freeze-dried state. The cartridge 300 may be formed of a plastic material that has excellent chemical resistance, is not hard, and is transparent. For example, the cartridge 300 may be formed of polypropylene (PP), polycarbonate (PC), acrylic, or the like.

The diagnostic module body 400 is detachably coupled to the cartridge 300, supplies heat of a certain temperature required for the nucleic acid amplification reaction to the cartridge 300 according to preset operating conditions, and changes the sample solution by the nucleic acid amplification reaction. It is possible to diagnose the presence or absence of a diagnosis target by detecting the color or fluorescence intensity of . Here, the operating condition may be set to a state in which the cartridge 300 is inserted into the diagnostic module body 400 and the sample solution is mixed with reagents in the cartridge 300 to complete preparation for performing the nucleic acid amplification reaction. .

The diagnostic module main body 400 may be controlled by the user terminal 2 and may communicate with the user terminal 2 to transmit diagnosis results for a diagnosis target. That is, in the integrated molecular diagnosis device 1 according to an embodiment of the present invention, procedures such as the user transferring the sample solution to the cartridge 300 can be omitted, and the user's intervention is minimized, from sample collection to nucleic acid detection. A preprocessing process of extraction and amplification and a diagnosis process can be implemented in one device.

The user terminal 2 may communicate with the integrated molecular diagnostic device 1 to control the operation of the integrated molecular diagnostic device 1 . The user terminal 2 may receive diagnosis results from the integrated molecular diagnosis device 1 and display them on the screen. Here, the diagnosis result may be displayed as negative or positive. In addition, the user terminal 2 may provide a screen displaying the required diagnosis time and diagnosis result. The user terminal 2 may transmit and store the diagnosis place, date, diagnosis time, etc. together with the diagnosis results in a database. Here, the database may be located inside or outside the user terminal 2 and may be managed by a separate server.

The user terminal 2 may be a computing device used by a user using the integrated molecular diagnosis device 1 . For example, the user terminal 2 may be a computing device such as a smart phone, tablet PC, desktop PC, laptop PC, etc., but is not limited thereto. An application that works with the integrated segmentation diagnosis device 1 may be installed in the user terminal 2 .

3 is a view for explaining the buffer tube shown in FIG. 1, and FIGS. 4a to 4c are views for explaining the opening and closing body shown in FIG.

Referring to FIG. 3 , the buffer tube 200 may include a tube body 210, an opening/closing body 220, and an inlet stopper member 230. The tube body 210 is formed in a cylindrical shape and includes an inner space accommodating a buffer solution. The upper end of the tube body 210 is opened by the opening/closing body 220 to accommodate the sampling tool 100 in the inner space.

Here, the upper end of the tube body 210 may be sealed with a sealing film (not shown), and the sealing film may be removed during a molecular diagnostic test. The tube body 210 may include a stepped 211 and a concave groove 213 formed on an outer circumferential surface to engage the cartridge 300 . Step 211 is formed to have a relatively larger diameter than the diameter of the tube body (210). The step 211 is engaged with the insertion hole 331 of the cartridge holder 330 when the tube body 210 is seated in the cartridge 300 can prevent the separation of the tube body 210.

The concave groove 213 is formed along the outer circumferential surface to have a relatively smaller diameter than the diameter of the tube body 210 . The concave groove 213 is formed at a position corresponding to the coupling protrusion 335 of the cartridge holder 330 and engaged with the coupling protrusion 335 . Accordingly, the tube body 210 is pressed while being inserted into the cartridge 300 by the elastic force of the elastic member 333 provided on the coupling protrusion 335 . Due to this, the buffer tube 200 may be fixed together with the cartridge 300 .

The opening and closing body 220 is coupled to the top of the tube body 210 to open and close the inner space of the tube body 210. As shown in FIGS. 4A to 4C , the opening and closing body 220 includes a concavo-convex pattern 221, a protruding jaw 223, a through hole 225, a plurality of ventilation holes 227, and a locking jaw 229. can do. The concavo-convex pattern 221 is formed on the upper surface of the opening and closing body 220, and minimizes the contact area of the user's hand during the opening and closing operation and the insertion of the buffer tube 200 into the cartridge 300 to prevent sample contamination. It can be prevented.

The protruding jaw 223 protrudes from the bottom surface of the opening and closing body 220 and is formed in an annular shape so that its outer circumferential surface is inserted into the inner circumferential surface of the tube body 210. The through hole 225 is formed through upper and lower surfaces corresponding to the central region of the opening and closing body 220 .

Each of the plurality of ventilation holes 227 is formed on the lower surface of the opening and closing body 220 between the side surface of the through hole 225 and the protruding jaw 223 . Each of the plurality of ventilation holes 227 may be formed spaced apart from each other by a predetermined distance.

The locking jaw 229 is bent inward from the side of the through hole 225 to support the inlet stopper member 230 . Here, the bent surface of the locking jaw 229 is deformed by the pressing force transmitted through the inlet stopper member 230 when the inlet stopper member 230 is moved, and the movement of the inlet stopper member 230 is stopped. In this state, the inlet stopper member 230 may be supported.

The inlet stopper member 230 is inserted into the through hole 225 of the opening and closing body 220 to seal the inner space of the tube body 210 . Here, the inlet stopper member 230 may be supported by the locking jaw 229 . The inlet stopper member 230 is pressed by the opening and closing operation of the diagnostic module body 400 and moves in the direction of the inner space of the tube body 210 to open the plurality of ventilation holes 227 . That is, the inlet stopper 230 forms an air inlet path through which air passes into the internal space by the opening and closing operation of the diagnostic module body 400 .

The inlet stopper member 230 may be formed of a material capable of passing air and blocking the sample solution (or buffer solution). That is, the inlet stopper member 230 is formed of a hydrophobic material so that the sample solution does not flow out even when the buffer tube 200 or the cartridge 300 is turned upside down, and has a certain length to perform the stopper function. can be formed

5 is a view for explaining an embodiment of the cartridge shown in FIG. 1, FIGS. 6A to 6C are views for explaining the cartridge body shown in FIG. 5, and FIGS. 7A and 7B are It is a drawing for explaining the cartridge holder shown in FIG. 5 .

Referring to FIG. 5 , the cartridge 300 may include a cartridge body 310 , a plurality of outlet stopper members 320 and a cartridge holder 330 . Here, the cartridge body 310 is formed in a plate shape including front and rear surfaces, as shown in FIGS. 6A to 6C, and includes a tube receiving body 311, an inlet 313, and a plurality of fluid channels 315. ), a plurality of reaction chambers 317 and a plurality of outlets 319.

The tube accommodating body 311 is formed in a protruding shape on the front surface of the cartridge body 310, and has an open upper end to include an inner space into which the buffer tube 200 is inserted. Here, the inner space may be formed in the same shape and size as the buffer tube 200 .

The tube receiving body 311 may include a coupling hole 311a and a punching member 311b. The coupling hole 311a may be formed through the front surface of the tube receiving body 311 at a position corresponding to the coupling protrusion 335 of the cartridge holder 330 .

The punching member 311b is formed on the support surface of the tube accommodating body 311, and can punch the bottom surface of the buffer tube 200 using pressure applied by an operation in which the buffer tube 200 is inserted. . The punching member 311b protrudes upward from the support surface of the tube receiving body 311 and may be formed to have a pointed end.

The inlet 313 is formed through the rear surface of the cartridge body 310 at the support surface of the tube receiving body 311 . The inlet 313 introduces the sample solution discharged from the bottom surface of the buffer tube 200 into each of the plurality of fluid channels 315 .

Each of the plurality of fluid channels 315 is formed on the rear surface of the cartridge body 310, and a sample solution may be transferred from the inlet 313 to the corresponding outlet 319 via the corresponding reaction chamber 317. Here, each of the plurality of fluid channels 315 may include a first flow path 315a and a second flow path 315b.

The first flow path 315a may be formed by branching from the inlet 313 into each of the plurality of reaction chambers 317 . The second flow path 315b may be formed between the corresponding reaction chamber 317 and the outlet 319 . The second flow path 315b may be formed in a zigzag curved shape in order to increase fluid resistance. Accordingly, the fluid resistance of the sample solution flowing through the second flow path 315b is increased so that the moving speed can be maintained constant.

Each of the plurality of reaction chambers 317 is formed on the rear surface of the cartridge body 310 and receives the sample solution transferred through each of the plurality of fluid channels 315 . Here, each of the plurality of reaction chambers 317 may contain reagents injected in advance, and may perform a nucleic acid amplification reaction with respect to the sample solution by receiving heat of a certain temperature from the diagnostic module body 400 . Each of the plurality of reaction chambers 317 may be formed to a size capable of accommodating a fixed amount of sample solution.

One embodiment of the present invention describes a case where the plurality of reaction chambers 317 are three as an example, but one embodiment of the present invention is not limited thereto, and the reaction chambers 317 are configured according to the number of target diagnosis subjects. may increase or decrease.

Each of the plurality of outlets 319 is formed on the front surface of the cartridge body 310 and is formed between the corresponding reaction chamber 317 and the tube receiving body 311 . That is, since each of the plurality of outlets 319 is positioned above the corresponding reaction chamber 317 , the sample solution may be maintained in a filled state in the plurality of reaction chambers 317 .

On the other hand, the cartridge body 310 according to an embodiment of the present invention further includes a sealing member (not shown) for sealing the inlet 313, the plurality of fluid channels 315 and the plurality of reaction chambers 317 on the rear side. can do. The sealing member is transparent and can be formed of a thin film.

Each of the plurality of outlet stopper members 320 is inserted into each of the plurality of outlets 319 . Each of the plurality of outlet stopper members 320 may pass air and block discharge of the sample solution flowing through each of the plurality of fluid channels 315 . Each of the plurality of outlet plug members 320 may be formed of a porous material, for example, polyethylene or porous hydrogel material. Accordingly, the sample solution may be maintained in a state where the flow in the plurality of fluid channels 315 is stopped without being discharged to the outside.

The cartridge holder 330 is coupled to the front of the cartridge body 310 and fixes the buffer tube 200 within the cartridge body 310 . As shown in FIGS. 7A and 7B , the cartridge holder 330 includes an internal space into which the cartridge body 310 is inserted, and includes an insertion hole 331, an elastic member 333 and a coupling protrusion 335. can do.

The insertion hole 331 is formed at a position corresponding to the upper end of the tube accommodating body 311 to expose the inner space of the tube accommodating body 311 . The elastic member 333 is a leaf spring and is formed on an inner surface facing the tube receiving body 311 . The elastic member 333 is elastically deformed by a sliding operation according to the insertion of the buffer tube 200 and provides elastic force to the coupling protrusion 335 .

The coupling protrusion 335 protrudes from the elastic member 333 at a position corresponding to the coupling hole 311a of the tube receiving body 311 and is inserted into the coupling hole 311a. The coupling protrusion 335 has an inclined inclined surface, and induces maximum elastic deformation of the elastic member 333 at the end of the inclined surface, thereby increasing the elastic restoring force according to the restoration of the elastic member 333 . When the coupling protrusion 335 reaches the concave groove 213 by a sliding operation according to the insertion of the buffer tube 200, it receives an elastic restoring force from the elastic member 333 and enters the concave groove 213 with a 'click' sound. jamming is fastened

That is, the coupling protrusion 335 makes it possible to know that the tube body 210 is completely coupled to the cartridge body 310 in place with a 'click' sound when engaged with the concave groove 213. Also, the coupling protrusion 335 couples the buffer tube 200 and the cartridge body 310 into one to prevent the buffer tube 200 from being separated again from the cartridge body 310 after one-time fastening.

8 is a block diagram for explaining the diagnostic module main body shown in FIG. 1, FIG. 9 is a block diagram for explaining the main body shown in FIG. 8, and FIG. 10 is a heat supply module shown in FIG. , and FIG. 11 is a diagram for explaining the detection module shown in FIG. 8 .

Referring to FIG. 8 , the diagnostic module body 400 includes a body 410, a heat supply module 420, a detection module 430, a power module 440, a detection module 450, and an integrated control module 460. can include The body 410 accommodates the cartridge 300, the heat supply module 420, the detection module 430, the power module 440 and the integrated control module 460. As shown in FIG. 9 , the body 410 may include a lower body 411 and an opening/closing body 413 . The lower body 411 may be formed in a rectangular shape and includes an internal space of a certain size. The lower body 411 may include an insertion hole 411a. The insertion hole 411a is formed on the upper surface of the lower body 411 and may be formed in a shape and size corresponding to the cartridge 300 in order to insert the cartridge 300 therein.

The opening and closing body 413 is coupled to the lower body 411 to open and close the inner space of the lower body 411 . The opening and closing body 413 may include a pressing member 413a. The pressing member 413a protrudes from the inner surface corresponding to the upper surface of the lower body 411 and may be formed at a position corresponding to the inlet stopper member 230 of the buffer tube 200 . The pressing member 413a may press and move the inlet stopper 230 .

The heat supply module 420 is detachably coupled to the cartridge 300 and is controlled by the integrated control module 460 to supply heat of a certain temperature required for the nucleic acid amplification reaction to each of the plurality of reaction chambers 317 .

As shown in FIG. 10 , the heat supply module 420 may include a heat conduction body 421 and a heating unit 423 . The heat conduction body 421 is coupled to the cartridge 300, and receives heat of a certain temperature from the heating means 423 and transfers it to the cartridge 300. The heat conduction body 421 may accommodate the cartridge 300 and the heating unit 423, and may include a cartridge insertion groove 421a, a plurality of first holes 421b, and a plurality of second holes 421c. .

The cartridge insertion groove 421a is formed at a position corresponding to the insertion hole 411a of the lower body 411, and the front, rear, and bottom of the cartridge 300 has an inner surface including a plurality of reaction chambers 317. It can be formed to contact the surface.

Each of the plurality of first holes 421b is formed through an inner surface of one side of the cartridge insertion groove 421a. Each of the plurality of first holes 421b may be formed at a position corresponding to each of the plurality of reaction chambers 317 .

Each of the plurality of second holes 421c is formed through the bottom surface of the cartridge insertion groove 421a. Each of the plurality of second holes 421c may be formed at a position corresponding to each of the plurality of reaction chambers 317 .

The heating means 423 is disposed in the heat conducting body 421 and generates heat of a certain temperature. The heating unit 423 may include a resistive heater, a thermoelectric element, and the like.

An embodiment of the present invention is not limited thereto, and the heat supply module 420 may further include a heat sink that dissipates heat from the heat conduction body 421 to the outside.

The detection module 430 is disposed adjacent to the heat supply module 420 and is controlled by the integrated control module 460 to irradiate light to each of the plurality of reaction chambers 317 and to each of the plurality of reaction chambers 317. Light passing through is detected to generate a detection signal.

As shown in FIG. 11 , the detection module 430 may include a plurality of light sources 431 and a plurality of light detectors 433 . Each of the plurality of light sources 431 may be controlled by the integrated control module 460 to radiate light to each of the plurality of reaction chambers 317 . Each of the plurality of light sources 431 may be formed of a light emitting diode (LED) or a laser diode (LD).

Here, each of the plurality of light sources 431 may be disposed adjacent to the plurality of second holes 421c of the heat conducting body 421 . Each of the plurality of light sources 431 may be disposed in a horizontal direction or a direction perpendicular to each of the plurality of light detectors 433 based on the cartridge 300 . An embodiment of the present invention illustrates a case where each of the plurality of light sources 431 is disposed in a direction perpendicular to each of the plurality of photodetectors 433, but the embodiment of the present invention is not limited thereto, Each of the plurality of light sources 431 may be disposed in a direction parallel to each of the plurality of light detectors 433 based on the cartridge 300 .

Preferably, when the photodetector 433 detects the color of the sample solution, the light source 431 may be disposed in a horizontal direction with the photodetector 433, and the photodetector 433 detects fluorescence of the sample solution. In this case, the light source 431 may be disposed in a direction perpendicular to the photodetector 433 .

Each of the plurality of photodetectors 433 may detect light passing through each of the plurality of reaction chambers 317 , generate a detection signal, and transmit the detected signal to the integrated control module 460 . Each of the plurality of photodetectors 433 may be disposed facing each of the plurality of reaction chambers 317 . Each of the plurality of photodetectors 433 may be disposed adjacent to the plurality of first holes 421b of the heat conducting body 421 . Here, each of the plurality of photodetectors 433 is a photodiode (PD), a photo multiplier tube (PMT), a phototransistor, or a charge-coupled device (CCD) or complementary metal-oxide semiconductor (CMOS) It may include an image sensor such as

The power module 440 may supply power to each of the heat supply module 420 , the detection module 430 and the integrated control module 460 . The power module 440 may include a battery, a power button, and a power terminal.

The sensing module 450 may sense the closed state of the body 410 to generate an open/close detection signal, and may sense the temperature of the heat supply module 420 to generate a temperature sensing signal. The sensing module 450 may include a plate spring member (not shown) supporting the opening/closing body 413 , a pressure sensor (not shown), and a temperature sensor (not shown).

The sensing module 450 may generate an opening/closing detection signal by detecting an elastic force generated from a leaf spring member during a closing operation of the opening/closing body 413 through a pressure sensor. Also, the sensing module 450 may sense the temperature of the heat supply module 420 through a temperature sensor and generate a temperature sensing signal.

The integrated control module 460 may determine whether a preset operating condition is satisfied according to the detection signal and the open/close detection signal transmitted from the detection module 430 . Specifically, the integrated control module 460 may determine whether to insert the cartridge 300 and transfer the sample solution into the plurality of reaction chambers 317 according to the detection signal. That is, the integrated control module 460 according to an embodiment of the present invention includes the detection module 430 as a sensor for determining whether the cartridge 300 is inserted into the body 410 and a sample in each reaction chamber 317. Operating conditions can be determined by using it as a sensor for determining whether a solution has been injected.

In addition, the integrated control module 460 may determine whether the body 410 is opened or closed according to the open/close detection signal. That is, the integrated control module 460 may determine that all operating conditions are satisfied when the cartridge 300 is inserted, the sample solution is transferred into the plurality of reaction chambers 317, and the body 410 is in a closed state. .

When the operating conditions are satisfied, the integrated control module 460 supplies heat at a constant temperature necessary for the nucleic acid amplification reaction to each of the plurality of reaction chambers 317 through the heat supply module 420 . The integrated control module 460 may constantly control the temperature of the heat supply module 420 according to the temperature detection signal.

When heat of a certain temperature is supplied to each of the plurality of reaction chambers 317, the integrated control module 460 detects a change in color or fluorescence intensity due to a nucleic acid amplification reaction from the sample solution of each of the plurality of reaction chambers 317 to diagnose. The presence or absence of an object can be diagnosed. The integrated control module 460 may communicate with the user terminal 2 to transmit a diagnosis result for a diagnosis target. The integrated control module 460 may be controlled by the user terminal 2 and may be implemented as a Printed Circuit Board (PCB) substrate.

12 is a flow chart illustrating a molecular diagnosis method according to an embodiment of the present invention, FIG. 13 is a diagram for explaining the movement of the inlet stopper of the buffer tube, and FIG. 14 is a flow path of a sample solution. 15A and 15B are diagrams for explaining diagnosis results.

Referring to FIG. 12 , first, a sample is taken from the user using the sampling tool 100 (step S110). Then, the sampling tool 100 is put into the tube body 210 of the buffer tube 200 (step S120). At this time, the sampling tool 100 is immersed in the buffer solution pre-injected into the buffer tube 200 . Then, the opening and closing body 220 is closed. At this time, since the inlet stopper member 230 is inserted into the through hole 225 of the opening and closing body 220, when the opening and closing body 220 is closed, the tube body 210 is sealed.

Then, shake the buffer tube (200). Then, the cell membrane of the sample collected through the sampling tool 100 is destroyed and nucleic acid is extracted (step S130). Accordingly, a sample solution in which the nucleic acid is mixed with the buffer solution is prepared.

In this state, the cartridge 300 is mounted on the diagnostic module main body 400 through the insertion hole 411a of the lower body 411 (step S140). Then, the buffer tube 200 is inserted into the tube receiving body 311 of the cartridge 300. At this time, the bottom surface of the tube body 210 is perforated by the perforation member (311b) (step S150). Then, the sample solution is discharged from the tube body 210. At this time, since the plurality of ventilation holes 227 are closed by the inlet stopper 230 of the buffer tube 200, the sample solution does not flow into the inlet 313.

When the opening and closing body 413 is closed in this state, as shown in FIG. 13 , the pressing member 413a of the opening and closing body 413 presses the inlet stopper 230 of the buffer tube 200, and the inlet stopper member 230 moves downward to open a plurality of ventilation holes 227 . Accordingly, an air inlet path A through which air is introduced into the inner space of the tube body 210 is formed.

Then, as shown in FIG. 14, the sample solution passes from the inlet 313 through the corresponding reaction chamber 317 by capillary force in a state in which air flows from both ends of the plurality of fluid channels 315. It moves along path B leading to the corresponding outlet 319 . At this time, since each of the plurality of outlets 319 is blocked by the corresponding outlet stopper 320, the sample solution is accommodated in the reaction chamber 317 with the flow stopped. In this way, the sample solution is transferred to the corresponding reaction chamber 317 through each of the plurality of fluid channels 315 (step 160).

At this time, the integrated control module 460 determines whether the operating condition is satisfied according to the detection signal and the opening/closing detection signal. For example, the integrated control module 460 may determine whether the cartridge 300 is inserted into the body 410 according to the detection signal, and determine whether a sample solution is injected into each reaction chamber 317. . In addition, the integrated control module 460 may determine whether the body 410 is opened or closed according to the open/close detection signal. Here, the integrated control module 460 is considered to satisfy all operating conditions when the cartridge 300 is inserted into the diagnostic module body 400 and the body 410 is closed and the sample solution is injected into each reaction chamber 317. can judge

In this way, when all operating conditions are satisfied, the integrated control module 460 supplies heat of a certain temperature to the plurality of reaction chambers 317 through the heat supply module 420 . Accordingly, a nucleic acid amplification reaction of the sample solution accommodated in each of the plurality of reaction chambers 317 is performed. At this time, the detection module 430 detects the color or fluorescence intensity of the sample solution to generate a detection signal and transmits it to the integrated control module 460 .

Then, the integrated control module 460 detects a change in color or fluorescence intensity due to a nucleic acid amplification reaction from the sample solution of each of the plurality of reaction chambers 317 according to the detection signal to diagnose the presence or absence of the diagnosis target.

For example, as shown in FIG. 15A , the integrated control module 460 may use a phenol red indicator (R) or a purple indicator (P) whose color is changed by a nucleic acid amplification reaction. When a color change (R1→R2 or P1→P2) due to a change in pH is detected before and after the nucleic acid amplification reaction in the included sample solution, it can be determined that the diagnosis target is present.

In addition, as shown in FIG. 15B , the integrated control module 460 may diagnose the presence or absence of a diagnosis target by detecting the fluorescence intensity of the sample solution through a nucleic acid amplification reaction. That is, the integrated control module 460 may determine that a diagnosis target exists when the fluorescence intensity of the sample solution increases as in (B), unlike in (A).

Then, the integrated control module 460 provides the diagnosis result for the diagnosis target to the user terminal 2 (step S170). Thereafter, the buffer tube 200 and the cartridge 300 may be sealed and disposed of.

Meanwhile, one embodiment of the present invention is not limited thereto, and in step S140, when the cartridge 300 is inserted into the diagnostic module body 400, diagnosis is made with the buffer tube 200 inserted into the cartridge 300. It can be inserted into the module body 400. That is, the buffer tube 200 may be inserted into the cartridge 300 first, and the cartridge 300 together with the buffer tube 200 may be inserted into the diagnostic module body 400.

16 is a diagram showing an integrated molecular diagnostic system according to another embodiment of the present invention, FIGS. 17A to 17C are diagrams for explaining the cartridge shown in FIG. 16, and FIG. 18 is a diagram showing the cartridge shown in FIG. It is a drawing shown to explain the diagnostic module body. 19 is a diagram for explaining the heat supply module shown in FIG. 18, FIGS. 20A and 20B are diagrams for explaining the detection module shown in FIG. 18, and FIG. 21 is a diagram shown in FIG. It is a diagram shown to explain the detection signal output from the detection module.

Referring to FIG. 16 , an integrated molecular diagnostic system according to another embodiment of the present invention may include an integrated molecular diagnostic device 3 and a user terminal 4 . The integrated molecular diagnostic device 3 may include a sampling tool 100', a buffer tube 200', a cartridge 300', and a diagnostic module body 400'. Here, since the sampling tool 100' and the buffer tube 200' have the same configurations as the sampling tool 100 and the buffer tube 200 according to an embodiment of the present invention, detailed descriptions thereof are omitted.

The cartridge 300' is the same as the cartridge 300 according to one embodiment of the present invention, but further includes a sensing means 340. Accordingly, the same components are described with the same reference numerals, and detailed descriptions are omitted for convenience of description. Here, the sensing unit 340 reacts with the hydrogen ions included in the sample solution of each of the plurality of reaction chambers 317 to detect the hydrogen ion concentration (pH). The sensing unit 340 may include a reference electrode 341 and a plurality of sensing electrodes 343 as shown in FIGS. 17A to 17C .

The reference electrode 341 is coupled to the rear surface of the cartridge body 310, and the reference electrode 341 covers the entire opening of the inlet 313, the plurality of fluid channels 315, and the plurality of reaction chambers 317. It can be formed in the form of a plate. That is, the reference electrode 341 according to an embodiment of the present invention may serve as a sealing member sealing the rear surface of the cartridge body 310 .

The reference electrode 341 includes an electrode terminal surface 341a extending downward from the bottom surface of the cartridge body 310, and is electrically connected to the diagnostic module body 400 through the electrode terminal surface 341a. can

The reference electrode 341 may contact the sample solution through a surface corresponding to the opening surface of each of the plurality of reaction chambers 317 . The reference electrode 341 has a constant reference potential in response to changes in the pH of the sample solution. The reference electrode 341 may be formed of a half-cell reactive material having pH stability and high reproducibility. For example, the reference electrode 341 may be formed of Ag/AgCl.

Here, the half-cell refers to a battery in which a potential difference occurs according to a half-reaction of oxidation or reduction. That is, the reference electrode 341 is a reducing electrode or an oxidizing electrode different from the plurality of sensing electrodes 343 when an oxidation or reduction reaction occurs in each of the plurality of sensing electrodes 343 according to the pH value of the sample solution. can operate as

Each of the plurality of sensing electrodes 343 is spaced apart from the reference electrode 341 and contacts the sample solution in the inner space of each of the plurality of reaction chambers 317 . Each of the plurality of sensing electrodes 343 may be formed through the cartridge body 310 in the inner space of the plurality of reaction chambers 317 . That is, each of the plurality of sensing electrodes 343 may be formed to be inserted into the inner space of each of the plurality of reaction chambers 317 from the bottom surface of the cartridge body 310 . Therefore, one end of each of the plurality of sensing electrodes 343 is disposed in the inner space of each of the plurality of reaction chambers 317 to contact the sample solution, and the other end is exposed on the bottom surface of the cartridge body 310 to diagnose module body. (400) and can be electrically connected.

Each of the plurality of sensing electrodes 343 has a sensing potential that changes according to a change in the pH of the sample solution. That is, both ends of each of the reference electrode 341 and the plurality of sensing electrodes 343 operate like a potential capacitor, and the sensing potential of each of the plurality of sensing electrodes 343 is changed based on the reference potential of the reference electrode 341. can Here, each of the plurality of sensing electrodes 343 may be formed of a metal oxide material that is sensitive to the hydrogen ion concentration (pH), such as ITO or SiO ₂ .

The diagnostic module body 400' is detachably coupled to the cartridge 300', supplies heat of a certain temperature required for the nucleic acid amplification reaction to the cartridge 300', and changes the hydrogen ion concentration by the nucleic acid amplification reaction with electricity. The presence or absence of the diagnosis target can be diagnosed by measuring the signal.

As shown in FIG. 18, the diagnostic module body 400' includes a body 410', a heat supply module 420', a detection module 430', a power module 440', a detection module 450 and An integrated control module 460' may be included. Here, since the body 410' and the power module 440' have the same configuration as the body 410 and the power module 440 according to an embodiment of the present invention, a detailed description thereof will be omitted.

The heat supply module 420' is detachably coupled to the cartridge 300' and controlled by the integrated control module 460' to supply heat of a certain temperature required for the nucleic acid amplification reaction to each of the plurality of reaction chambers 317. supply

As shown in FIG. 19 , the heat supply module 420' may include a heat conduction body 421' and a heating means 423'. The heat conduction body 421', into which the cartridge 300' is inserted, receives heat of a certain temperature from the heating means 423' and transfers it to the cartridge 300'.

The heat conduction body 421' accommodates the cartridge body 310 and the heating means 423, and may include a cartridge insertion groove 421a' and first and second connector insertion holes 421b' and 421c'. there is. The cartridge insertion groove 421a' is formed at a position corresponding to the insertion hole 411a of the lower body 411, and the front and rear surfaces of the cartridge body 310 have an inner surface including a plurality of reaction chambers 317. And it may be formed to contact the bottom surface. The cartridge insertion groove 421a' may have a bottom surface having a step corresponding to the electrode terminal surface 341a of the reference electrode 341 .

The first connector insertion hole 421b' is formed through the bottom surface of the cartridge insertion groove 421a', and the second connector insertion hole 421c' is spaced apart from the first connector insertion hole 421c' by a predetermined distance. It may be formed through the bottom surface of the cartridge insertion groove (421a').

The heating unit 423' is disposed in the heat conducting body 421' and is controlled by the integrated control module 460' to generate heat at a constant temperature. The heating means 423' may include a resistive heater, a thermoelectric element, and the like. An embodiment of the present invention is not limited thereto, and the heat supply module 420' may further include a cooling means for discharging heat generated from the heat conducting body 421' to the outside.

The detection module 430' is electrically connected to the detection unit 340' of the cartridge 300' to supply a reference voltage of a predetermined level to the reference electrode 341, and to detect the detection voltage of each of the plurality of detection electrodes 343. is detected to generate a plurality of detection signals. The detection module 430' may transmit the detection signal to the integrated control module 147'.

As shown in FIG. 20A, the detection module 430' may include a reference electrode connector 431', a plurality of sensing electrode connectors 433', a reference voltage supply unit 435, and a hydrogen ion concentration detection unit 437. can The reference electrode connector 431' is inserted into the first connector insertion hole 421b' of the heat conducting body 421' and contacts the reference electrode 341.

The reference electrode connector 431' includes an insertion groove 431a into which the electrode terminal surface 341a of the reference electrode 341 is inserted, and may make surface contact with the reference terminal surface 341a. An embodiment of the present invention is not limited thereto, and the reference electrode connector 431' may be bent in an 'L' shape to contact the electrode terminal surface 341a of the reference electrode 341.

Each of the plurality of sensing electrode connectors 433' is inserted into each of the plurality of second connector insertion holes 421c' of the heat conducting body 421' and makes contact with each of the plurality of sensing electrodes 343. Each of the plurality of sensing electrode connectors 433' may contact each of the plurality of sensing electrodes 343 on the bottom surface of the cartridge body 310.

The reference voltage supplier 435 supplies a voltage of a certain potential to the reference electrode 341 through the reference electrode connector 431'.

The hydrogen ion concentration detector 437 is electrically connected to each of the plurality of sensing electrodes 343 through each of the plurality of sensing electrode connectors 433', and detects a sensing potential of each of the plurality of sensing electrodes 343 to obtain a plurality of generate a detection signal.

The hydrogen ion concentration detector 437 may include a plurality of non-inverting operational amplifiers (APs). As shown in FIG. 20B, each of the plurality of non-inverting operational amplifiers (AP) has a non-inverting input terminal (+) connected to each of the plurality of sensing electrode connectors 433' and an inverting input terminal (-) to which a ground voltage is applied. and an output terminal for outputting the detection signal Vout.

The sensing potential (Vs) of the sensing electrode 343 changes by ΔVs based on the reference potential (Vr) according to the change in the pH of the sample solution (SS), and the non-inverting operational amplifier (AP) changes the sensing potential The amount of change in (Vs) can be output as a detection signal (Vout). That is, the hydrogen ion concentration detector 437 may generate the detection signal Vout by detecting a change in the pH of the sample solution in the plurality of reaction chambers 317 .

For example, as shown in FIG. 21 , it can be seen that the voltage level of the detection signal Vout changes as shown in (C) according to the change of the hydrogen ion concentration (pH). An embodiment of the present invention is not limited thereto, and materials of each of the reference electrode 341 and the plurality of sensing electrodes 343 may be exchanged, and in this case, the detection signal Vout has a hydrogen ion concentration as shown in (D). It can show a trend change in the opposite direction to a change in (pH).

The sensing module 450' may detect the temperature of the heat supply module 420', generate a temperature sensing signal, and transmit the temperature sensing signal to the integrated control module 460'. The sensing module 450' may include a temperature sensor.

The integrated control module 460' supplies heat at a constant temperature necessary for the nucleic acid amplification reaction to each of the plurality of reaction chambers 317 through the heat supply module 420'. Here, the integrated control module 460' may constantly control the temperature of the heat supply module 420' according to the temperature detection signal.

When heat of a certain temperature is supplied to each of the plurality of reaction chambers 317, the integrated control module 460' diagnoses the existence of a diagnosis target according to a plurality of detection signals. The integrated control module 460' determines whether the hydrogen ion concentration of each sample solution in the plurality of reaction chambers 317 changes according to a plurality of detection signals, and determines that the diagnosis target exists when the hydrogen ion concentration changes. can

The integrated control module 460' may communicate with the user terminal 4 to transmit diagnosis results for a diagnosis target. The integrated control module 460 ′ may be controlled by the user terminal 4 and may be implemented as a PCB (Printed Circuit Board) substrate.

As described above, the integrated molecular diagnosis device 1 according to an embodiment of the present invention can be implemented as a single device including pre-processing of the collected sample and diagnosis results. Therefore, diagnosis results can be obtained conveniently and quickly. In addition, by obtaining a diagnosis result in an optical method or an electrochemical sensor method, the device can be implemented in a small and simple manner, so that it can be used for on-site diagnosis. In addition, since the diagnosis result can be checked in the user terminal 2, convenience can be improved.

Claims

a buffer tube into which a sampling tool for collecting a sample is inserted and generating a sample solution containing nucleic acid extracted from the collected sample;

a cartridge coupled to the buffer tube to receive the sample solution, and carrying out a nucleic acid amplification reaction by transferring the sample solution to a reaction chamber through a fluidic channel; and

An integrated molecular diagnostic device comprising a diagnostic module main body detachably coupled to the cartridge to supply heat of a certain temperature to the reaction chamber and detecting the nucleic acid amplification reaction to diagnose the presence or absence of a diagnostic target.
The method of claim 1, wherein the buffer tube

An integrated molecular diagnostic device for extracting the nucleic acid by destroying the cell membrane of the sample by a shaking operation in which a buffer solution is pre-injected and the sample collection tool is inserted.
The method of claim 2, wherein the buffer tube

a tube body accommodating the sampling tool and including an inner space in which the buffer solution is pre-injected;

an opening and closing body coupled to the tube body to open and close the inner space; and

and an inlet stopper inserted through the opening/closing body and selectively introducing air into the inner space of the tube body by moving by a pressing force applied from the diagnostic module body.
The method of claim 3, wherein the opening and closing body

a protruding jaw that protrudes from the lower surface and is formed in an annular shape so that the outer circumferential surface is inserted into the inner circumferential surface of the tube body;

a through hole formed between the upper surface and the lower surface and into which the inlet stopper is inserted; and

An integrated molecular diagnostic device comprising a plurality of ventilation holes formed on the lower surface between the side surface of the through hole and the protruding jaw and spaced apart from each other.
According to claim 3,

The diagnostic module body includes a pressing member formed at a position corresponding to the inlet stopper member,

The pressing member applies the pressing force to the inlet stopper member by opening and closing the diagnostic module body.
The method of claim 1, wherein the buffer tube

An integrated molecular diagnostic device formed of a plastic material containing at least one of polypropylene and polycarbonate.
The method of claim 1, wherein the cartridge

A cartridge body formed in the form of a plate including front and rear surfaces; and

An integrated molecular diagnostic device including a cartridge holder coupled to the front surface of the cartridge body.
The method of claim 7, wherein the cartridge body

a tube accommodating body formed on the front surface of the cartridge body and including an inner space into which the buffer tube is inserted;

an inlet formed through the rear surface of the cartridge body at the support surface of the tube receiving body in contact with the bottom surface of the buffer tube;

an outlet formed on the front surface of the cartridge body and disposed between the inlet and the reaction chamber;

the fluid channel formed on the rear surface of the cartridge body and transporting the sample solution from the inlet to the outlet;

the reaction chamber formed in the fluid channel on the rear surface of the cartridge body to accommodate the sample solution, to contain a pre-injected reagent, and to perform the nucleic acid amplification reaction by receiving heat of the constant temperature; and

An integrated molecular diagnostic device comprising an outlet plug member inserted into the outlet to pass air and block the sample solution.
The method of claim 8, wherein the fluid channel

a first flow path formed from the inlet to the reaction chamber; and

An integrated molecular diagnostic device comprising a second flow path formed from the reaction chamber to the outlet.
10. The method of claim 9, wherein the second flow path

An all-in-one molecular diagnostic device formed in a zigzag curved shape.
The method of claim 8, wherein the outlet stopper member

An integrated molecular diagnostic device formed of at least one of porous polyethylene and porous hydrogel.
The method of claim 8, wherein the tube receiving body

a coupling hole formed on the front; and

An integrated molecular diagnostic device comprising a piercing member formed on the support surface to pierce a bottom surface of the buffer tube.
13. The method of claim 12, wherein the cartridge holder

an elastic member formed on an inner surface facing the tube receiving body and elastically deformed by a sliding operation according to insertion of the buffer tube; and

An integrated molecular diagnostic device comprising a coupling protrusion formed to protrude from the elastic member at a position corresponding to the coupling hole and having an inclined inclined surface.
According to claim 13,

The buffer tube includes a concave groove formed on an outer circumferential surface at a position corresponding to the coupling hole,

The integrated molecular diagnostic device of
8. The method of claim 7, wherein the cartridge

An integrated molecular diagnostic device comprising a sensing means for sensing the hydrogen ion concentration of the sample solution in the reaction chamber.
16. The method of claim 15, wherein the sensing means

a reference electrode contacting the sample solution and having a constant reference potential in response to a change in hydrogen ion concentration of the sample solution; and

An integrated molecular diagnostic device including a sensing electrode spaced apart from the reference electrode, contacting the sample solution, and having a sensing potential that changes according to a change in the hydrogen ion concentration of the sample solution.
17. The method of claim 16, wherein the reference electrode

An all-in-one molecular diagnostic device formed of Ag/AgCl.
17. The method of claim 16, wherein the reference electrode

Integrated molecular diagnosis comprising an electrode terminal surface formed in a plate shape covering the opening surface of the reaction chamber, coupled to the cartridge body, and extending from the bottom surface of the cartridge body to electrically contact the diagnostic module body. Device.
17. The method of claim 16, wherein the sensing electrode

An integrated molecular diagnostic device formed from the inner space of the reaction chamber through the cartridge body.
The method of claim 15, wherein the diagnostic module body

a detection module electrically connected to each of the reference electrode and the detection electrode and generating a detection signal by detecting a detection potential of the detection electrode; and

An integrated molecular diagnostic device comprising an integrated control module for diagnosing the existence of the diagnosis target by determining a change in the concentration of hydrogen ions in the sample solution according to the detection signal.
The method of claim 1, wherein the cartridge

An integrated molecular diagnostic device comprising at least one of polypropylene, polycarbonate, and acryl and formed of a transparent plastic material.
The method of claim 1, wherein the diagnostic module body

A body including a lower body including an inner space of a predetermined size and an insertion hole into which the cartridge is inserted on an upper surface thereof, and an opening and closing body coupled to the lower body to open and close the inner space;

a heat supply module disposed in the inner space of the body, detachably coupled to the cartridge through a cartridge insertion groove formed at a position corresponding to the insertion hole, and supplying heat of the constant temperature to the reaction chamber;

a detection module disposed in the inner space of the body to irradiate light into the reaction chamber and detect a color or fluorescence intensity of the sample solution to generate a detection signal; and

Integrated molecular diagnosis including an integrated control module disposed in the inner space of the body and diagnosing the existence of the diagnosis target by determining a change in color or fluorescence intensity of the sample solution by the nucleic acid amplification reaction from the detection signal Device.
23. The method of claim 22, wherein the detection module

a light source for radiating light to the reaction chamber; and

An integrated molecular diagnostic device comprising a photodetector generating the detection signal by detecting the color or fluorescence intensity of the sample solution irradiated with the light.
The method of claim 22, wherein the integrated control module

An integrated molecular diagnostic device that communicates with a user terminal and transmits a diagnosis result for the diagnosis target to the user terminal.
Collecting a sample using a sampling tool;

preparing a sample solution by extracting nucleic acids from the sample when the sample collection tool is inserted into the buffer tube;

discharging the sample solution from the buffer tube in a state where the buffer tube is mounted on the cartridge and the cartridge is inserted into the main body of the diagnostic module, and transferring the sample solution to a reaction chamber through a fluid channel formed in the cartridge;

performing a nucleic acid amplification reaction by applying heat of a certain temperature to the reaction chamber; and

A molecular diagnosis method using an integrated molecular diagnosis device comprising the step of diagnosing the presence or absence of a diagnosis target by detecting the nucleic acid amplification reaction through the diagnosis module body.
The method of claim 25, wherein preparing the sample solution

Molecules using an integrated molecular diagnostic device comprising the step of extracting the nucleic acid by destroying the cell membrane of the sample by shaking the buffer tube in a state where the sample collection tool is immersed in the buffer solution injected in advance into the buffer tube diagnostic method.
The method of claim 25, wherein the step of transferring the sample solution

Discharging the sample solution from the buffer tube by puncturing the bottom surface of the buffer tube; and

A molecular diagnosis method using an integrated molecular diagnosis device comprising the step of introducing air into an inlet and an outlet connected to both ends of the fluid channel.
28. The method of claim 27, wherein introducing air into the inlet

and opening a plurality of ventilation holes formed in an opening and closing body for opening and closing the buffer tube by a closing operation of the diagnostic module body after the cartridge is inserted.
According to claim 28,

The plurality of ventilation holes are closed by an inlet stopper member penetrating the opening and closing body, and are opened when a pressing member formed on the diagnostic module body presses the inlet stopper member by the closing operation of the diagnostic module body. Molecular diagnosis method using a diagnostic device.
The method of claim 25, wherein diagnosing whether or not the diagnosis target exists

irradiating light into the reaction chamber;

detecting the color or fluorescence intensity of the sample solution; and

A molecular diagnosis method using an integrated molecular diagnosis device comprising the step of determining whether the color or fluorescence intensity is changed before and after the nucleic acid amplification reaction.
The method of claim 25, wherein the step of determining whether the diagnosis target exists

generating a detection signal by detecting a sensing potential that changes based on a reference potential according to the hydrogen ion concentration of the sample solution in the reaction chamber; and

A molecular diagnosis method using an integrated molecular diagnosis device comprising the step of determining a change in the concentration of hydrogen ions in the sample solution according to the detection signal.
32. The method of claim 31, wherein the reference potential is

A molecular diagnosis method using an integrated molecular diagnosis device, which is a potential level of a reference electrode formed in a plate shape covering an opening surface of the reaction chamber and contacting the sample solution.
33. The method of claim 32, wherein the reference electrode

A molecular diagnostic method using an integrated molecular diagnostic device formed of Ag/AgCl.
33. The method of claim 32, wherein the sense potential is

A molecular diagnosis method using an integrated molecular diagnosis device, which is a potential level of a sensing electrode formed from an inner space of the reaction chamber and passing through the cartridge while being spaced apart from the reference electrode and in contact with the sample solution in the reaction chamber.
35. The method of claim 34, wherein the sensing electrode

A molecular diagnosis method using an integrated molecular diagnosis device formed of a metal oxide material containing at least one of ITO and SiO 2 .