Diagnostic kit for nucleic acid extraction, amplification and analysis

Abstract

The present invention relates to a diagnostic kit for nucleic acid extraction, amplification, and analysis, the diagnostic kit comprising: a kit main body in which a row of upper-open cells are formed so as to accommodate supplied reagents and samples, a sample supply channel for inputting a sample is formed on the front side of a first compartment among the plurality of compartments, and the first compartment is connected to the first compartment, the first compartment is configured to accommodate a plurality of magnetic beads therein and to convey the plurality of magnetic beads from the first compartment to the rearmost compartment through a magnetic member from the outside, and the plurality of magnetic beads are conveyed from the first compartment to the rearmost compartment through the magnetic member in order to convey the magnetic beads. Each partition plate among the plurality of compartments is provided with a structure in which the compartments are communicated with one another; a reagent supply part which is inserted through the upper part of the opening of the cartridge main body; and a reaction chamber coupled to a side surface of the cartridge main body to perform nucleic acid amplification.

Description

TECHNICAL FIELD

[0001] The present application relates to a device for extracting high-purity nucleic acids from high-viscosity biological samples such as sputum, whole blood, etc., and more particularly, to a diagnostic cartridge for nucleic acid extraction, amplification, and analysis. BACKGROUND

[0002] In modern times, with the development of biotechnology, it is possible to interpret the cause of a disease at the genetic level. As a result, there is an increasing demand for manipulation and biochemical analysis of biological samples to cure or prevent human diseases.

[0003] In general, diagnostic analysis and testing of biological samples refers to detecting or measuring specific indicator substances from biological samples such as blood, urine, saliva, etc., thereby determining whether or not a disease is infected. Such diagnostic analysis and testing of biological samples is not only used for diagnostic purposes such as etiological confirmation, treatment, prevention, prediction, etc., but also widely used in many fields such as targeted new drug development, forensic medicine, environmental hormone detection, etc.

[0004] Polymerase Chain Reaction (PCR) testing, which is widely known as a method of biological sample diagnostic analysis and testing, is a test that amplifies the DNA of bacteria and thereby confirms whether or not bacteria are infected.

[0005] In order to perform the PCR test, a pre-treatment step of extracting and amplifying genetic material such as DNA, RNA, etc., i.e., nucleic acids, from the biological sample is necessary.

[0006] Meanwhile, in addition to the diagnosis of diseases, techniques for extracting and analyzing nucleic acids from biological samples or cell-containing samples are also required in many fields such as new drug development, pre-testing of viral or bacterial infection, forensic medicine, etc.

[0007] Conventional nucleic acid extraction devices require separate devices for each process (concentration, purification), and need to be transferred to another device after completing one process, so it takes a long time.

[0008] In addition, since the test sample can flow out to the outside of the cartridge, there is a need to develop a diagnostic cartridge that can perform diagnostic analysis and testing more safely.

[0009] On the other hand, U.S. Patent No. 6,374,684 proposes a method to solve the problem of low detection efficiency due to the long process of conventional nucleic acid extraction devices. In this patent, a plurality of branch flow paths are provided at the piston head, the piston head is rotated and can directly suck the reagent in the cavity, and mixing is performed in the internal space of the piston.

[0010] However, U.S. Patent No. 6,374,684 requires the generation of multiple branched flow paths inside the piston head, and there is a problem of increased manufacturing costs of the piston, and thus there is a need to develop a more inexpensive and simple structure of a cartridge for nucleic acid extraction. SUMMARY

[0011] PROBLEMS TO BE SOLVED BY THE INVENTION To this end, the present invention was developed to solve the above problems, and aims to provide a diagnostic cartridge for nucleic acid extraction, amplification, and analysis, which can safely perform diagnosis without the detection sample flowing outside the cartridge, and can be conveniently provided at a lower cost.

[0012] TECHNICAL SOLUTION TO THE PROBLEMS According to one embodiment of the present invention for achieving the above object, a diagnostic cartridge for nucleic acid extraction, amplification, and analysis includes a cartridge main body having a plurality of compartments formed in a row with an upper portion open to accommodate supplied reagents and samples, a sample supply passage for supplying a sample being formed on a front side of a first compartment among the plurality of compartments and connected to the first compartment, the first compartment being configured to accommodate a plurality of magnetic beads therein, the plurality of magnetic beads being transported from the first compartment to a last side compartment by a magnetic member from the outside, each partition plate between the plurality of compartments having a structure in which the compartments are connected to each other; a reagent supply part containing at least a reagent (or a washing solution) and being insertedly provided through the open upper portion of the cartridge main body; and a reaction chamber provided at one side of the cartridge main body to perform nucleic acid amplification.

[0013] According to the present invention, a portion of each partition plate between the plurality of compartments is lower than the periphery, so that nucleic acids adsorbed on the magnetic beads in the compartments can be transported to a side compartment by a magnetic member operating outside the cartridge main body, and in order to supply at least a reagent (or a washing solution) to each compartment in the cartridge main body, the reagent supply part has a plurality of reagent chambers formed in a row with lower ends closed by a sealing film, each of the reagent chambers having a reagent (and a washing solution) accommodated therein, and a puncture needle or a puncture plate for puncturing the sealing film when the reagent supply part descends is built in the inside of each compartment, wherein the plurality of reagent chambers can have an inclination of at least 45 degrees, so that the reagent can smoothly flow into the inside of the main body after puncturing.

[0014] According to the present invention, the diagnostic kit further includes: a reagent discharge tube connected to the inlet of the flow path in the reaction chamber and the kit body, for discharging nucleic acid extracted from the kit body into the reaction chamber; and a syringe connected to the outlet of the flow path in the reaction chamber, thereby drawing the extracted nucleic acid into the reaction chamber from the reagent discharge tube; the reagent supply unit has a sample injection port formed on the front side of the first reagent chamber corresponding to the first compartment of the plurality of compartments of the kit body, corresponding to the sample supply channel, for injecting a sample into the first compartment.

[0015] According to the present invention, the reagent supply unit has a nucleic acid extraction port on its rear end side corresponding to the last compartment of the plurality of compartments of the main body for extracting purified nucleic acid from the last compartment of the main body. When extracting nucleic acid through the nucleic acid extraction port, a pipette or syringe can be used. In order to deliver the reagent containing the nucleic acid extracted from the sample to the reaction chamber, the reagent discharge tube has an inverted U-shaped cross-section structure connecting the last compartment of the plurality of compartments of the main body and the flow path inlet of the reaction chamber, and grooves can be formed on both sides for embedding and fixing to the side cutout of the main body.

[0016] According to the present invention, the reaction chamber may have engaging protrusions corresponding to the slots formed on the left and right sides of its front end, so that one end of its horizontal surface can be embedded and fixed into a mounting groove formed on one side of the main body, or embedded and combined into a slot provided on the upper left and right sides of a bracket formed on the side of the main body. Alternatively, the reaction chamber may be integrally formed with the box body.

[0017] According to the present invention, in order to discharge the nucleic acid extracted from the box body into the reaction chamber, the bottom surface of the last side compartment and the flow path inlet of the reaction chamber can be formed at the same height, so that the last side compartment of the box body and the flow path of the reaction chamber are interconnected. In addition, the diagnostic box also includes a predetermined aspiration mechanism, which generates vacuum pressure through a predetermined aspiration stroke to move the nucleic acid from the box body along the flow path within the reaction chamber. This aspiration mechanism can consist of a cylinder integrally formed with the side of the box body and a piston that performs the aspiration stroke along the interior of the cylinder.

[0018] According to the present invention, in order to fully mix the nucleic acids and reagents of the sample in the first compartment among the plurality of compartments of the box body, at least one ultrasonic generator may be provided in close contact with an aluminum film attached to the lower end of the body to provide ultrasonic waves. In addition, in order to unwind double-stranded DNA into single strands in a polymerase chain reaction process, the reaction chamber may also include at least one ultrasonic generator in close contact with an aluminum film that seals at least a portion of the reaction chamber to provide ultrasonic waves.

[0019] According to another aspect of the present invention, a diagnostic kit for nucleic acid extraction, amplification, and analysis includes: a kit body with an open upper portion and multiple compartments; and a reagent supply section having a row of multiple cavities for receiving reagents or samples, each cavity having a side wall inclined at 45 degrees; in the kit body, each of the multiple compartments has a puncture needle for piercing a sealing film adhered to one side of the reagent supply section when the reagent supply section is installed on the open upper portion of the kit body; at least a portion of each of the multiple compartments has a 45-degree inclination to allow the reagents or samples inside the reagent supply section to flow more smoothly into the interior of the kit body; the diagnostic kit further includes a reaction chamber disposed on one side of the kit body for performing nucleic acid amplification; in the multiple compartments, a portion of the partitions between the compartments is configured to have a height lower than a reference height.

[0020] Invention Effects As can be seen from the above technical features, the diagnostic kit according to the present invention can be applied not only to molecular diagnostics but also to immunodiagnostics. In particular, inside the kit body, magnetic components are used to transfer microbeads adsorbed with nucleic acids, so the test sample will not flow out of the kit, thereby enabling more safe diagnostic operations. Attached Figure Description

[0021] Figures la to Id This is a perspective view showing the appearance of the diagnostic box of the present invention and the composition of its various components.

[0022] Figure 2 This is an enlarged cross-sectional view of the reagent dispensing tube and syringe combined with the main body of the box in Figure 1.

[0023] Figure 3a and Figure 3b This is a partial perspective view showing the operational state of the diagnostic box of the present invention.

[0024] Figure 4 is a structural diagram showing the composition and connection of the main body of the box and the reaction chamber in Figure 1.

[0025] Figure 5 This is a schematic diagram illustrating the operation of the diagnostic box of the present invention.

[0026] Figure 6 This is a perspective view showing another embodiment of the diagnostic box of the present invention.

[0027] Figure 7a and Figure 7b It is shown Figure 6 The diagnostic kit features a three-dimensional view of the shape and structure of the main body of the kit.

[0028] Figure 8a and Figure 8c It is shown Figure 6A three-dimensional diagram showing the shape and structure of the reagent supply section of the diagnostic kit.

[0029] Figure 9a and Figure 9b This shows a reagent supply unit installed. Figure 6 A partial perspective view of the initial state of the diagnostic box.

[0030] Figures 10a to 10c This indicates when the reagent supply section descends. Figure 6 A 3D diagram showing the state changes of the diagnostic kit.

[0031] Figure 11a and Figure 11b It is used for explanation Figure 6 A cross-sectional illustration of the movement principle and path of reagents and nucleic acids within the diagnostic kit.

[0032] Figure 12a and Figure 12b This shows the method used for extraction. Figure 6 A diagram showing the usage status of purified DNA in a diagnostic kit.

[0033] Figure 13a and Figure 13b This is a perspective view showing the shape and structure of the main body of the diagnostic box according to another embodiment of the present invention.

[0034] Figure 14a and Figure 14b This is a perspective view showing the deformed internal structure of the diagnostic box body shown in Figure 13.

[0035] Figure 15a and Figure 15b This is a partial perspective view showing the initial state of the diagnostic box in Figure 13.

[0036] Figures 16a to 16b This is a perspective view showing the state change of the diagnostic box in Figure 13 as the reagent supply section descends.

[0037] Figure 17 This is a cross-sectional illustration used to explain the movement principle and path of reagents and nucleic acids within the diagnostic kit shown in Figure 13.

[0038] Figure 18 This is an illustrative diagram showing the arrangement of a POCT device for emitting ultrasound waves into a reaction chamber installed in a diagnostic box according to an embodiment of the present invention. Detailed Implementation

[0039] The present invention will now be described in more detail with reference to the accompanying drawings and embodiments.

[0040] In the following embodiments, except for the parts necessary to explain the invention, the illustrations and descriptions are omitted. Throughout the specification, the same and similar components are referred to by the same reference numerals, and detailed descriptions thereof are omitted and will not be repeated.

[0041] Figure 1 to Figure 2 The present invention illustrates the configuration of a diagnostic kit for nucleic acid extraction, amplification, and analysis. The diagnostic kit includes: a kit body 100 (hereinafter also referred to as the "body"), which is a quadrilateral shell with an open top for extracting nucleic acids from a sample; and a reaction chamber 200, which is coupled to the side of the kit body to perform nucleic acid amplification.

[0042] A reagent supply unit 300 is mounted on the upper part of the box body 100 through an opening. Below the reagent supply unit 300, a row of multiple compartments 111, 112, 113, 114, 115, ... (see reference) are formed inside the body 100 to accommodate the supplied reagents and samples. Figure 5 ).

[0043] Multiple compartments 111, 112, 113, 114, and 115 are enclosed by the upper part of the reagent supply unit 300. The first compartment 111, arranged sequentially, has a sample supply channel 101 for sample insertion formed on its front side and connected to the first compartment 111. Multiple magnetic beads 102 are built into the first compartment 111. Preferably, each compartment 111, 112, 113, 114, and 115 has a structure with partially interconnected upper sides, allowing the magnetic beads 102 to be transported from the first compartment 111 to the last compartment via a predetermined magnetic component 103 (in this embodiment, the fifth compartment 115 is illustratively shown and described as the last compartment).

[0044] To supply reagents (and washing liquid) to the compartments 111, 112, 113, 114, and 115 within the main body 100, a row of reagent cavities 311, 312, 313, 314, and 315 with lower puncture points are formed on the reagent supply unit 300 corresponding to each compartment. Each reagent cavity 311, 312, 313, 314, and 315 contains a puncture needle (not shown) for piercing the capsule containing the reagent (and washing liquid). In this invention, the reagent cavities 311, 312, 313, 314, and 315 of the reagent supply unit 300 may have an inclined surface of approximately 45 degrees to allow the reagent flowing from the reagent capsule (not shown) to flow more smoothly into the interior of the main body 100. The reagent supply unit 300 has a sample injection port 301 for injecting a sample into the first reagent chamber 311 on the front side of the first reagent chamber 311 corresponding to the first compartment 111 of the main body 100, corresponding to the sample supply channel 101.

[0045] The reaction chamber 200 is used to receive reagents containing nucleic acids from the main body 100 using the vacuum pressure of the syringe 400 and to perform nucleic acid amplification, and is laterally bound and fixed to the side of the main body 100. In order to fix the reaction chamber 200, a support 141 for supporting the lower end of the reaction chamber 200 is formed on the side of the main body 100, and a slot 142 corresponding to the engaging protrusions 220 formed on both sides of the front end of the reaction chamber 200 is formed on the upper left and right sides of the support 141.

[0046] The reaction chamber 200 contains multiple cavities and a flow path 210 through the probe (see reference). Figure 4b The end of syringe 400 is connected to the outlet 211 of the flow path 210 (hereinafter referred to as the "flow path outlet"), and the reagent discharge tube 500 extending from the last side compartment 115 within the main body is connected to the inlet 212 of the flow path (hereinafter referred to as the "flow path inlet") (see reference). Figure 3b ).

[0047] The syringe 400 can be detachably mounted to the mounting portion 120 formed on the side of the body 100 to create pressure for drawing fluid so that the reagent can be filled into the reaction chamber 200.

[0048] The reagent discharge tube 500 is used to deliver reagents containing nucleic acids extracted from the sample to the reaction chamber 200. It has an inverted U-shaped cross-sectional structure that connects the last side compartment 115 of the main body 100 and the flow path inlet 212 of the reaction chamber 200. It may have grooves 510 on both sides so that it can be embedded and fixed to the side cutouts 130 of the main body 100.

[0049] Reference Figure 4a To ensure thorough mixing of the nucleic acid and reagents in the sample, ultrasound can be supplied to the first compartment 111. For this purpose, a first ultrasound generator 600 (see reference) can be installed in the lower part of the first compartment 111. Figure 3a It is tightly attached to the aluminum film 150 sealed at the lower end of the main body 100.

[0050] In addition, in order to unwind double-stranded DNA into single strands in the polymerase chain reaction process, ultrasonic waves can be supplied to the reaction chamber 200. For this purpose, a second ultrasonic generator 700 can be provided at the lower end of the reaction chamber 200 (see reference). Figure 3b It is set in close contact with the sealed aluminum film 230.

[0051] The diagnostic box of the present invention having the configuration described above can be used as follows: Figure 5 As shown in the diagram.

[0052] First, when the collected sample is injected through the sample injection port 301 of the reagent supply unit 300, the sample flows into the first compartment 111 through the sample supply channel 101 of the main body 100. The reagent capsule (not shown) is punctured through the first reagent chamber 311 of the reagent supply unit 300, filling the first compartment 111 with reagent. Additionally, multiple magnetic beads 102 are inserted into the first compartment 111, and the magnetic beads 102 are moved from the outside using a magnetic component 103 to pulverize the sample and disrupt the cells, thereby thoroughly mixing the reagent and sample. Through this process, components flowing out of the cells of the sample can be adsorbed onto the surface of the microbeads 102.

[0053] In the configuration shown in this invention, the second reagent chamber 312, the third reagent chamber 313, and the fourth reagent chamber 314 of the reagent supply unit 300 can be supplied with washing liquid for cleaning by puncturing the capsule. The washing liquid flowing down from the second, third, and fourth reagent chambers 312, 313, and 314 respectively fills the interior of the second compartment 112, the third compartment 113, and the fourth compartment 114.

[0054] In this state, the magnetic beads 102 inside the first compartment 111 are moved sequentially to the second compartment 112, the third compartment 113 and the fourth compartment 114 by the magnetic component 103, and foreign matter on the surface of the magnetic beads 102 is removed by washing.

[0055] Additionally, the fifth reagent chamber 315 of the reagent supply unit 300 can supply an elution solution for nucleic acid extraction by puncturing the capsule. The elution solution flowing from the fifth reagent chamber 315 fills the interior of the last side compartment, namely the fifth compartment 115. At this time, when the washed magnetic beads 102 inside the fourth compartment 114 are moved into the fifth compartment 115 by the magnetic component 103, nucleic acid is extracted from the surface of the magnetic beads 102 by the elution solution.

[0056] The extracted nucleic acid is drawn into the reaction chamber 200 through the inverted U-shaped reagent discharge tube 500 using the vacuum pressure of the syringe 400 outside the main body 100 during the suction stroke. After flowing along the flow path 210, the nucleic acid is amplified.

[0057] The diagnostic kit of the present invention, which operates in this way, uses a magnetic component inside the kit body to transfer microbeads adsorbed with nucleic acids, so that the test sample will not flow out of the kit, thus providing the advantage of being able to perform diagnostic operations more safely.

[0058] The following is for reference Figure 6 Figure 13 illustrates another embodiment of the diagnostic kit for nucleic acid extraction, amplification, and analysis of the present invention.

[0059] The diagnostic kit of the present invention for nucleic acid extraction, amplification and analysis, such as Figure 6As shown, it includes: a box body 100' (hereinafter referred to as the "body") with an open upper part for extracting nucleic acid from a sample; a reagent supply unit 300' inserted through the open upper part of the body 100' and placed on the upper inner side of the body 100'; and a reaction chamber 200' combined with the side of the body 100' and performing nucleic acid amplification.

[0060] Figure 7a and Figure 7b Show Figure 6 The shape and structure of the main body of the box shown are as follows: the upper part of the main body 100' is open for mounting the reagent supply unit 300'; multiple slots 104a and 104b for accommodating the reagent supply unit 300' are formed at certain intervals along the upper part of the front and rear walls. In addition, below the reagent supply unit 300' which is placed in the slots 104a and 104b, multiple compartments 111, 112, 113, 114, 115, ... are formed in a row inside the main body 100' for accommodating the supplied reagents and samples.

[0061] In this embodiment, through Figures 7a to 7b Five compartments 111, 112, 113, 114, and 115 are schematically shown. The fifth compartment, i.e., the fifth compartment 115, will be referred to as the "last side compartment" below.

[0062] Multiple compartments 111, 112, 113, 114, and 115 are enclosed by the upper part of the reagent supply unit 300'. Among the sequentially arranged compartments 111, 112, 113, 114, and 115, the front side of the first compartment 111 has a sample supply channel 101 for sample insertion, which is connected to the first compartment 111. Multiple magnetic beads 102 (see reference) are built into the first compartment 111. Figure 11a Preferably, the upper part of the partition 105 of each compartment 111, 112, 113, 114, 115 is open, having a structure in which the compartments 111, 112, 113, 114, 115 are interconnected, so that a magnetic bead 102 can be conveyed from the first compartment 111 to the last side compartment 115 by a predetermined magnetic component 103 (magnet). Inventively, this interconnection structure can be achieved by providing a recess (connecting recess) 108, which is formed by at least reducing the height of a portion of the partition 105 located between each compartment 111, 112, 113, 114, 115.

[0063] Figures 8a to 8c Show Figure 6 The diagnostic kit has a reagent supply section with a specific shape and structure. Figure 9a and Figure 9b A partial perspective view shows the reagent supply unit installed. Figure 6 The initial state of the diagnostic kit, according toFigure 8b and Figure 8c It can be seen that the reagent supply section 300' has a row of reagent chambers 311, 312, 313, 314, 315 corresponding to the number of compartments 111, 112, 113, 114, 115, which are used to supply reagents (or washing liquid) to each compartment 111, 112, 113, 114, 115 in the main body 100'.

[0064] These reagent chambers 311, 312, 313, 314, and 315 are formed at an angle downwards and rearwards from the interior of the reagent supply section 300', and have a structure with an open lower portion and a closed upper portion through the upper surface of the reagent supply section 300'. Each reagent chamber 311, 312, 313, 314, and 315 can pass through a predetermined thin film material 310a (a sealing film; see reference) while filled with reagent (or washing liquid). Figure 9b Seal the lower part of the opening.

[0065] On the one hand, in order to pierce the sealing film 310a and supply the internal reagents (or washing solutions) into the compartments 111, 112, 113, 114, 115 of the main body 100', in the main body 100', each compartment 111, 112, 113, 114, 115 has a puncture needle 107 that protrudes sharply upwards and corresponds to the sealing film 310a of each reagent chamber 311, 312, 313, 314, 315 (see reference). Figure 7b ).

[0066] Reference Figure 8b and Figure 9b The reagent cavities 311, 312, 313, 314, and 315, filled with reagent (or washing solution), have a structure that extends downwardly and obliquely from the closed upper surface, as shown above. A sealing film 310a is adhered to the lower opening to block the outflow of the reagent (or washing solution). Preferably, as shown in the figure, each reagent cavity 311, 312, 313, 314, and 315 has an inclination of approximately 45 degrees towards the front of the main body 100' inside the reagent supply section 300'. As the reagent cavities 311, 312, 313, 314, and 315 have an inclination of approximately 45 degrees, the lower ends of each reagent cavity 311, 312, 313, 314, and 315, sealed by the sealing film 310a, also have an inclination of approximately 45 degrees from the horizontal direction (see reference). Figure 8c Thus, with reagent chambers 311, 312, 313, 314, and 315 each having a 45-degree inclination angle, when the sealing film 310a is pierced, the reagent gathers and flows along the inclination surface in a point-to-point direction, thereby preventing residual reagent from remaining in the chamber (see reference). Figure 8c ).

[0067] Alternatively, preferably, at least a portion of the rear wall surface 106 of each compartment 111, 112, 113, 114, 115 of the main body 100' has an inclination angle of about 45 degrees, which allows the reagent supplied at an inclination angle of about 45 degrees to flow smoothly down from the reagent chambers 311, 312, 313, 314, 315.

[0068] In this embodiment, Figure 8a This shows a front perspective view of the reagent supply unit. Figure 8b The rear perspective view of the reagent supply section is shown. Preferably, at the end of the reagent supply section 300' in one direction, a sample injection port 301 for injecting a sample into the first compartment 111 inside the main body 100' is formed in the front direction of the first reagent chamber 311 among the reagent chambers 311, 312, 313, 314, and 315.

[0069] The sample injection port 301 extends downward to be close to the sample supply channel 101 of the main body 100'. In order to inject the sample, an opening and closing door 302 that can be opened and closed can be provided at the inlet of the sample injection port 301. The opening and closing door 302 can be configured to slide along the front end upper surface of the reagent supply section 300' with opposing guide members 303 at a certain interval. For this purpose, guide grooves 302a for embedding in the guide members 303 are formed on both sides of the opening and closing door 302.

[0070] Additionally, at the other end of the reagent supply section 300', preferably, on the upper surface of the reagent supply section 300' on the same line as the fifth reagent chamber 315 (in this embodiment, the fifth reagent chamber 315 is illustratively shown and described as the "last side reagent chamber"), a nucleic acid extraction port 305 for extracting nucleic acid from the interior of the last side compartment 115 within the main body 100' penetrates the interior of the reagent supply section 300' and extends downward in correspondence with the last side compartment 115.

[0071] To extract nucleic acid, an opening and closing door 306 can be provided at the entrance of the nucleic acid extraction port 305. This opening and closing door 306 can be configured to slide along the upper surface of the last reagent chamber 315 with opposing guide members 307 at certain intervals. For this purpose, guide grooves 306a for embedding in the guide members 307 are formed on both sides of the opening and closing door 306.

[0072] The aforementioned opening and closing doors 302 and 306 may have a handle protruding from the upper surface to facilitate easy opening and closing by the user. Preferably, stop protrusions 303a and 307a for each opening and closing door 302 and 306 are formed at the front side of the guide members 303 and 307 that slide on the opening and closing doors 302 and 306, i.e., at the end of the sample injection port 301 side and the nucleic acid extraction port 305 side. Fitting grooves 302b and 306b are formed at the front end of the opening and closing doors 302 and 306 for respectively embedding and fixing to the stop protrusions 303a and 307a (see reference). Figure 8b and Figure 8c ).

[0073] Engaging protrusions 308 are formed on the front and rear surfaces of the reagent supply section 300', which are used to mount the reagent supply section 300' in an engaged state onto the main body 100'. The reference numeral "304" not explained in the figure indicates the reduced space of the reagent supply section 300'.

[0074] The reaction chamber 200' is used to perform nucleic acid amplification by receiving reagents containing nucleic acids from the body 100 using vacuum pressure based on the syringe 400. In this embodiment, the reaction chamber 200' is embedded horizontally into the side of the body 100'. Figure 6 The explanation will focus on the combination structure shown.

[0075] In order to install the reaction chamber 200', a mounting groove 142 for embedding and fixing the reaction chamber 200' is formed on the side of the main body 100', preferably on the side of the rearmost compartment 115 of the main body 100'.

[0076] The reaction chamber 200' will be described below with a structure in which one end of the horizontal plane is embedded and fixed to the mounting groove 142 formed on the side of the main body 100', but it is not limited to this and may also be an embodiment as described above. Figure lb As shown, the engaging protrusions 220 are formed on both sides of the front end of the reaction chamber 200, and the slots 142 are formed on the upper left and right sides of the bracket 141 formed on the side of the main body 100, so that when the reaction chamber 200 is engaged, the engaging protrusions 220 are embedded in the slots 142.

[0077] The internal structure of the reaction chamber 200' can be in the same manner as described in the above embodiment. That is, as... Figure 4b As shown, the reaction chamber 200 has multiple chambers and a flow path 210 through the probe. The end of the syringe 400 is connected to the outlet 211 of the flow path 210 (hereinafter referred to as the "flow path outlet"). The reagent discharge tube 500, extending from the last side compartment 115 inside the main body 100', is connected to the inlet 212 of the flow path (hereinafter referred to as the "flow path inlet"). (Ref) Figure 5 ).

[0078] In this embodiment, the syringe 400 can be detachably mounted on the bracket 120 formed on the side of the main body 100' to create a suction pressure, i.e., a negative suction pressure, so that the reagent can be filled into the reaction chamber 200'.

[0079] In addition, in this embodiment, the diagnostic kit also includes a reagent discharge tube 500 connecting the last side compartment 115 of the main body 100 and the reaction chamber 200'. The reagent discharge tube 500 is used to deliver reagents containing nucleic acids extracted from the sample to the reaction chamber 200', and has the following structure: a discharge channel 501 with an inverted U-shaped cross-section structure having a flow path inlet 212 connecting the last side compartment 115 of the main body 100 and the reaction chamber 200' formed inside, and grooves 510 formed on both sides so as to be able to be embedded and fixed to the side cutout 130 of the main body 100.

[0080] Figures 10a to 10c The diagram shows the state change of the diagnostic cartridge when the reagent supply section 300', mounted on the upper part of the main body in Figure 9, is pressed down to lower it. (Refer to...) Figure 9a and Figure 9b In the initial state, the reagent supply section 300' is located on the upper inner side of the main body 100'. In this state, the engaging protrusions 308 protruding from the front and rear surfaces of the reagent supply section 300' are engaged in the upper engaging grooves 104a formed on the front and rear surfaces of the main body 100' to support the reagent supply section 300'. In addition, at this time, it is known that the puncture needles 107 formed in each compartment 111, 112, 113, 114, 115 of the main body 100' are kept at a certain distance from the sealing film 310a pasted on the lower end face of the reagent chambers 311, 312, 313, 314, 315 downwards.

[0081] After that, as Figure 10a and Figure 10b As shown, when the reagent supply section 300' is pressed down so that the reagent chambers 311, 312, 313, 314, and 315 descend, the engaging protrusions 308 protruding from the front and rear surfaces of the reagent supply section 300' also descend downward and are engaged in a position in the lower side grooves 104b formed on the front and rear surfaces of the main body 100' to support the reagent supply section 300'.

[0082] And, at this time, such as Figure 10cAs shown, as reagent chambers 311, 312, 313, 314, and 315 descend, the sealing film 310a sealing the lower end face of reagent chambers 311, 312, 313, 314, and 315 is punctured by the puncture needles 107 formed in each compartment 111, 112, 113, 114, and 115 of the main body 100'. Consequently, the reagent (or washing solution) filled in reagent chambers 311, 312, 313, 314, and 315 can flow down along the rear wall surface 106 inside the main body 100' and be supplied to the interior of each compartment 111, 112, 113, 114, and 115.

[0083] The diagnostic box of the present invention having the configuration described above can be used as follows: Figure 11a and Figure 11b As shown in the diagram.

[0084] Figure 11a and Figure 11b It is used for explanation Figure 6 A cross-sectional illustration of the movement principle and path of reagents and nucleic acids in the diagnostic kit. First, a sample is collected from the human body and injected into the sample injection port 301 of the reagent supply unit 300'. The sample then flows into the first compartment 111 through the sample supply channel 101 of the main body 100'.

[0085] At this time, when the reagent supply section 300' located on the upper inner side of the main body 100' is pressed down to lower it, the sealing film 310a sealing the lower end face of the reagent chambers 311, 312, 313, 314, and 315 is punctured by the puncture needles 107 of each compartment 111, 112, 113, 114, and 115 within the main body 100' (see reference). Figure 10c As a result, reagents (or washing solutions) flow down from each reagent chamber 311, 312, 313, 314, 315 and fill the interior of each compartment 111, 112, 113, 114, 115.

[0086] Multiple magnetic beads 102 are introduced into the first compartment 111, and the magnetic beads 102 are moved from the outside using a magnetic component 103 (magnet) to pulverize the sample and destroy the cells, thereby thoroughly mixing the reagent and the sample. Through this process, components flowing out of the cells of the sample can be adsorbed onto the surface of the microbeads 102.

[0087] In this embodiment, the second, third, and fourth reagent chambers 312, 313, and 314 of the reagent supply unit 300' contain washing liquid for cleaning. Due to the descent of the aforementioned reagent supply unit 300', the sealing film 310a is punctured by the puncture needle 107, thereby filling the second, third, and fourth reagent chambers 312, 313, and 314 with washing liquid. In this state, the magnetic beads 102 inside the first chamber 111 are moved sequentially to the second, third, and fourth chambers 112, 113, and 114 by the magnetic component 103, and foreign matter on the surface of the magnetic beads 102 is removed through a washing process based on the washing liquid.

[0088] In this embodiment, the reagent chamber 315 at the rear end of the reagent supply unit 300' contains an elution solution for nucleic acid extraction. Similarly, due to the descent of the reagent supply unit 300', the sealing film 310a is punctured by the puncture needle 107, allowing the elution solution in the rear reagent chamber 315 to fill the interior of the rear compartment 115, i.e., the fifth compartment 115, within the main body 100'. At this time, when the washed magnetic beads 102 inside the fourth compartment 114 are moved to the rear compartment 115 using the magnetic component 103, nucleic acid (DNA) is extracted from the surface of the magnetic beads 102 using the elution solution.

[0089] At this point, the extracted nucleic acid (DNA) is transferred from the top of the precipitated magnetic beads 102 to the upper part of the reagent level. Using the vacuum pressure based on the suction stroke of the syringe 400 located on the side of the main body 100, it can be discharged into the reaction chamber 200' through the inverted U-shaped discharge channel 501 inside the reagent discharge tube 500. After the nucleic acid (DNA) flows into the reaction chamber 200', it flows along the flow path 210 while nucleic acid amplification occurs.

[0090] Typically, PCR amplification efficiency is affected by a variety of factors. Common factors include time, temperature, the amount and type of polymerase, DNA quantity, dNPT, and Mg2+. When the purified DNA is sent to the reaction chamber 200', it is necessary to separate the DNA from the magnetic beads 102 that capture it from the extraction step. For this purpose, in this embodiment, a magnet 103 is used to capture the microbeads 102.

[0091] In addition, when the purified DNA is sent to the reaction chamber 200', the remaining reagent that is not adsorbed on the magnet will flow into the reaction chamber 200'. In this embodiment, in order to prevent this phenomenon, preferably, the inlet of the inverted U-shaped discharge channel 501 in the reagent discharge tube 500 is located at a water level higher than that of the microbeads 102, so as to draw in DNA at a water level higher than that of the precipitated microbeads 102.

[0092] The diagnostic kit of the present invention, having the structure and operation described above, can be applied not only to molecular diagnostics but also to immunodiagnostics. In particular, since magnetic beads adsorbed with nucleic acids are transferred using magnets inside the kit body, the test sample will not flow out of the kit, thus providing the advantage of enabling safer diagnostic operations.

[0093] Figure 12a and Figure 12b An example of use for extracting purified DNA from the diagnostic kit of the present invention is shown, wherein a pipette P (or a syringe) can be used to extract the purified DNA from the last side compartment, i.e., the fifth compartment 115.

[0094] Typically, kits for traditional point-of-care testing (POCT) devices are designed for single-use testing only. Therefore, the purified DNA obtained through the internal processes cannot be retrieved from the kit. Consequently, if a test fails in a POCT device, the purified DNA cannot be reused, requiring a new sample to be collected, causing inconvenience.

[0095] However, according to the present invention, the purified DNA can be extracted by opening the opening and closing door 306 and inserting the pipette P into the last side compartment 115 through the nucleic acid extraction port 305.

[0096] Therefore, even if a POCT test fails, since the extracted DNA is still from the same patient, a second test can be performed without the need for repeated sample collection as in traditional methods. Of course, the extracted DNA can also be used for other purposes besides this.

[0097] Figure 13a and Figure 13b This illustrates yet another embodiment of the diagnostic box of the present invention. According to this embodiment, the reaction chamber 200” can be integrally formed with the box body 100”, without the need for FIG1 and Figure 6 The bracket 141 and slot 142 shown in the configuration have their internal flow path 210 and the last side compartment 115' of the box body connected to each other. The syringe 400' can be in a state where its end is integrally combined with the flow path outlet 211 of the reaction chamber 200". The surface of the cylinder 410 is integrally formed with the box body 100".

[0098] In particular, in this embodiment, the diagnostic kit can be seen from Figure 1 and Figure 6 In this configuration, the reagent discharge tube 500 is removed, and the bottom height of the last side compartment 115' of the main body 100" is raised to the same height as the reaction chamber 200". Therefore, the flow path 210 between the last side compartment 115' and the reaction chamber 200" no longer requires the reagent discharge tube 500 (Figure 1 and...). Figure 6 They can also be interconnected.

[0099] Figure 14a and Figure 14b The enlarged view shows the deformed internal structure of the diagnostic box body in Figure 13. Figure 7b In the configuration, the puncture needles 107, which have an upwardly protruding shape corresponding to the sealing films 310a of each reagent chamber 311, 312, 313, 314, and 315, can be configured as plate-shaped puncture plates 107' in this embodiment. Preferably, such puncture plates 107' may include at least a pair of plates spaced apart from each other and arranged side by side.

[0100] Figures 15 and 16 are partial perspective views showing the state changes of the diagnostic box before and after the reagent supply section is lowered. In the initial state shown in Figure 15, the reagent supply section 300' is located on the upper inner side of the main body 100'. In this state, the engaging protrusions 308' protruding from the left and right surfaces of the reagent supply section 300' are engaged and placed on the engaging steps 104c formed on the left and right surfaces of the main body 100' to support the reagent supply section 300'. In addition, at this time, it can be seen that the puncture plates 107' formed in each compartment 111, 112, 113, 114, 115' of the main body 100' are kept at a certain distance from the sealing film 310a pasted on the lower end face of the reagent chambers 311, 312, 313, 314, 315 downwards.

[0101] After that, as Figure 16a As shown, when the reagent supply section 300' is pressed downwards to lower the reagent chambers 311, 312, 313, 314, and 315, as... Figure 16b As shown, the sealing film 310a of the lower end face of the sealed reagent chambers 311, 312, 313, 314, 315 is punctured by the puncture plate 107' formed in each compartment 111, 112, 113, 114, 115' of the main body 100". As a result, the reagent (or washing liquid) filled in the reagent chambers 311, 312, 313, 314, 315 can flow down along the rear wall surface 106 inside the main body 100" and be supplied to the interior of each compartment 111, 112, 113, 114, 115'.

[0102] Figure 17This is a cross-sectional illustration illustrating the movement principle and path of reagents and nucleic acids within the diagnostic kit shown in Figure 13. After a sample collected from a human body is inserted into the first compartment 111 through the sample injection port 301 of the reagent supply unit 300', as shown in Figure 16, pressing down on the reagent supply unit 300' fills each compartment 111, 112, 113, 114, and 115' with reagents (or washing solution). Then, multiple magnetic beads 102 are inserted into the first compartment 111, and the magnetic beads 102 are moved from the outside using a magnetic component 103 (magnet) to pulverize the sample and disrupt the cells, thereby thoroughly mixing the reagents and sample. Through this process, components flowing out from the cells of the sample can be adsorbed onto the surface of the microbeads 102.

[0103] In this embodiment, the second, third, and fourth reagent chambers 312, 313, and 314 of the reagent supply unit 300' contain washing liquid for cleaning. Due to the descent of the aforementioned reagent supply unit 300', the sealing film 310a is punctured by the puncture plate 107', thereby filling the second, third, and fourth reagent chambers 312, 313, and 314 with washing liquid. In this state, the magnetic beads 102 inside the first chamber 111 are moved sequentially to the second, third, and fourth chambers 112, 113, and 114 by the magnetic component 103, and foreign matter on the surface of the magnetic beads 102 is removed through a washing process based on the washing liquid.

[0104] In this embodiment, the reagent chamber 315 at the rear end of the reagent supply unit 300' contains an elution solution for nucleic acid extraction. Similarly, due to the descent of the reagent supply unit 300', the sealing film 310a is punctured by the puncture plate 107', allowing the elution solution in the rear reagent chamber 315 to fill the interior of the rear compartment 115' within the main body 100'. At this time, when the washed magnetic beads 102 inside the fourth compartment 114 are moved to the rear compartment 115' using the magnetic component 103, nucleic acid (DNA) is extracted from the surface of the magnetic beads 102 using the elution solution.

[0105] At this time, the extracted nucleic acid (DNA) can be discharged into the reaction chamber 200' by the vacuum pressure based on the suction stroke of the syringe 400 integrated with the flow path outlet 211 of the reaction chamber. After flowing into the reaction chamber 200', the nucleic acid (DNA) moves along the flow path 210 and undergoes nucleic acid amplification.

[0106] Figure 18The diagram shows the internal configuration and usage of a POCT testing device for emitting ultrasound waves into a reaction chamber installed in a diagnostic kit according to an embodiment of the present invention. In order to unwind double-stranded DNA into single strands in a polymerase chain reaction process, ultrasound waves can be provided to the reaction chamber 200'. For this purpose, a metal film such as an aluminum film 230 is used to seal the reaction section inside the reaction chamber 200' containing DNA-containing reagents, and an ultrasound generator 700' is attached tightly to the underside of the aluminum film 230.

[0107] To denature and inactivate most of the DNA, this ultrasonic generator 700 raises the temperature to 95°C. If the temperature of the denaturation and inactivation regions can be reduced or eliminated during DNA amplification, the time in traditional PCR amplification processes involving repeated temperature cycles can be significantly shortened.

[0108] The foregoing has described various embodiments of the present invention, but the content described above is merely an example of some preferred embodiments of the present invention and is not limited thereto, unless otherwise specified in the appended claims. Therefore, those skilled in the art should understand that many changes, modifications, and equivalent substitutions can be made to the present invention within the scope of the claims without departing from the technical spirit and essence of the present invention.

Claims

1. A diagnostic kit for nucleic acid extraction, amplification, and analysis, comprising: The box body has a row of multiple compartments inside to hold supplied reagents and samples. The upper sides of the multiple compartments are open to allow for the supply of reagents. A sample supply channel for inserting a sample into the first compartment is formed on the front side of the first compartment. A reagent supply unit, containing at least a reagent (or washing solution), is inserted through the upper part of the opening of the main body of the box; as well as A reaction chamber, located on one side of the main body of the box, is used to perform nucleic acid amplification.

2. The diagnostic kit according to claim 1, characterized in that, The partitions between the multiple compartments of the box body are partially lower than the periphery, so that nucleic acids adsorbed on magnetic beads in the compartments can be transported to the side compartments by magnetic components that operate outside the box body.

3. The diagnostic kit according to claim 1, characterized in that, The reaction chamber is integrally formed with the main body of the box.

4. The diagnostic kit according to claim 1, characterized in that, Each compartment of the main body of the box contains one or more puncture needles or puncture plates, which are used to puncture the sealing film at the lower end of the reagent supply section when the reagent supply section descends.

5. The diagnostic kit according to claim 1, characterized in that, In the diagnostic kit, in order to discharge the nucleic acid extracted from the kit body into the reaction chamber, the bottom surface of the last side compartment and the flow path inlet of the reaction chamber are made at the same height, so that the last side compartment of the kit body and the flow path of the reaction chamber are interconnected. The diagnostic kit also includes a predetermined inhalation mechanism that generates a vacuum pressure through a predetermined inhalation stroke to move nucleic acids from the kit body along a flow path within the reaction chamber.

6. The diagnostic kit according to claim 1, characterized in that, In order to supply reagents (or washing liquid) to at least each compartment within the main body of the box, the reagent supply section is configured to contain reagents (and washing liquid) and is formed with a row of multiple reagent chambers corresponding to each compartment of the main body of the box, the lower ends of which are sealed by a sealing film, and the lower ends of the multiple reagent chambers have an inclination of at least 45 degrees.

7. The diagnostic kit according to claim 6, characterized in that, The reagent supply unit has a sample injection port on the front side of the first reagent chamber, which corresponds to the first compartment among the multiple compartments of the box body, and is formed in accordance with the sample supply channel for injecting a sample into the first compartment.

8. The diagnostic kit according to claim 6, characterized in that, The reagent supply unit has a nucleic acid extraction unit on the rear side corresponding to the last compartment among the multiple compartments of the box body. The nucleic acid extraction unit is used to extract purified nucleic acid from the last compartment in the box body. A pipette or syringe is used to extract nucleic acid through the nucleic acid extraction unit.

9. The diagnostic kit according to claim 1, characterized in that, In order to fully mix the nucleic acids and reagents of the sample in the first compartment among the multiple compartments of the box body, at least one ultrasonic generator is also included, which is closely disposed with respect to the aluminum film attached to the lower end of the box body to provide ultrasonic waves.

10. The diagnostic kit according to claim 1, characterized in that, In order to unwind double-stranded DNA into single strands in a polymerase chain reaction process, the reaction chamber further includes at least one ultrasonic generator disposed in close contact with at least a portion of an aluminum film sealing the reaction chamber to provide ultrasonic waves.

11. A diagnostic kit for nucleic acid extraction, amplification, and analysis, comprising: The main body of the box has an open top for extracting nucleic acid from samples. A reagent supply unit is inserted into the upper part of the opening of the main body of the box; A reaction chamber, integrally formed on the side of the main body of the box, is used to perform nucleic acid amplification; as well as An inhalation mechanism is connected to move the extracted nucleic acid from the cartridge body to the reaction chamber, and is connected to the flow path outlet in the reaction chamber. The nucleic acid from the cartridge body is filled into the reaction chamber using vacuum pressure based on the inhalation stroke.

12. The diagnostic kit according to claim 11, characterized in that, In multiple compartments, the partitions between the multiple compartments are formed by a height that is a certain portion lower than the reference height.

13. The diagnostic kit according to claim 11, characterized in that, The inhalation mechanism consists of a cylinder integrally formed with the main body of the box and a separately provided piston.

14. The diagnostic kit according to claim 11, characterized in that, The box body also includes a pipette or syringe as an extrusion mechanism.

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