Diagnostic cartridges for nucleic acid extraction, amplification, and analysis.

Abstract

The present invention relates to a diagnostic cartridge for nucleic acid extraction, amplification, and analysis, the diagnostic cartridge comprising: a plurality of open-topped compartments arranged in a row inside for accommodating supplied reagents and samples; a sample supply path for introducing a sample is formed in front of the first compartment and connected to the first compartment; the first compartment contains a plurality of magnetic beads and is configured to transport the plurality of magnetic beads from the first compartment to the final compartment by a magnetic member from the outside; each partition wall between the plurality of compartments has a structure that communicates between the compartments for transporting the magnetic beads; a reagent supply unit is inserted and placed through the open top of the cartridge body; and a reaction chamber is coupled to the side of the cartridge body and performs nucleic acid amplification.

Description

Technical Field

[0001] The present invention is for extracting high-purity nucleic acids from highly viscous biological samples such as sputum and whole blood, and more particularly relates to a diagnostic cartridge for nucleic acid extraction, amplification, and analysis.

Background Art

[0002] In modern times, the development of biotechnology has made it possible to interpret the causes of diseases at the gene level. As a result, the requirements for the manipulation of biological samples and biochemical analysis for treating or preventing human diseases are gradually increasing. [[ID=I3]]

[0003] Generally, a diagnostic analysis test for a biological sample is a test for determining the presence or absence of a disease infection by detecting or measuring a specific indicator substance from a biological sample such as blood, urine, saliva, etc. Such a diagnostic analysis test for a biological sample is widely used not only for diagnostic purposes such as confirmation of the cause of disease, treatment, prevention, and prognosis, but also in various fields such as customized new drug development, forensic medicine, and environmental hormone detection.

[0004] The PCR (Polymerase Chain Reaction) test, which is widely known as a method for the above-mentioned diagnostic analysis test of biological samples, is a test for confirming the presence or absence of bacterial infection by amplifying a biological gene using bacterial DNA.

[0005] In order to perform the above-mentioned PCR (Polymerase Chain Reaction) test, a pretreatment step of extracting and amplifying nucleic acids, which are genetic materials such as DNA and RNA, from the biological sample is essential.

[0006] In addition to disease diagnosis, technologies for extracting and analyzing nucleic acids from samples containing biological samples and cells are required in various fields such as new drug development, pre-examination of the presence or absence of virus or bacterial infection, and forensic medicine.

[0007] Conventional nucleic acid extraction systems require a separate device for each processing step (concentration, purification), and the process is time-consuming because the samples must be moved to the next device after each step is completed.

[0008] Furthermore, because samples could leak outside the cartridge, there was a need to develop a diagnostic cartridge that would allow for safer diagnostic analysis and testing.

[0009] On the other hand, to solve the problem of low detection efficiency due to the long processing time of conventional nucleic acid extraction devices, U.S. Patent No. 6,374,684 has been proposed. In this patent, multiple branched channels are created in the piston head, so that the piston head rotates and directly draws in the reagents in the chamber and mixes them in the internal space of the piston.

[0010] However, in the case of U.S. Patent No. 6,374,684, it is necessary to create multiple branched flow channels inside the piston head, which has the potential to increase the manufacturing cost of the piston. Therefore, there is a need to develop a cheaper and simpler cartridge structure for nucleic acid extraction. [Overview of the project] [Problems that the invention aims to solve]

[0011] Therefore, the present invention was developed to solve the aforementioned problems, and aims to provide a diagnostic cartridge for nucleic acid extraction, amplification, and analysis that can safely perform diagnosis without the sample leaking out of the cartridge, and can be provided easily and at a lower cost. [Means for solving the problem]

[0012] According to one embodiment of the present invention for achieving the above objective, a diagnostic cartridge for nucleic acid extraction, amplification, and analysis is provided, the diagnostic cartridge comprising: a plurality of open-topped compartments arranged in a row inside for accommodating supplied reagents and samples; a sample supply path for introducing a sample is formed in front of the first compartment and connected to the first compartment; the first compartment contains a plurality of magnetic beads inside and is configured to transport the plurality of magnetic beads from the first compartment to the final compartment by a magnetic member from the outside; each partition wall between the plurality of compartments has a structure that communicates between the compartments for transporting the magnetic beads; a reagent supply section containing at least a reagent (or wash solution) and inserted and placed through the open top of the cartridge body; and a reaction chamber provided on one side of the cartridge body for nucleic acid amplification.

[0013] According to the present invention, each partition wall between the plurality of compartments can be formed to be lower in height than the surrounding area so that nucleic acids attached to magnetic beads in the compartment are transported to the adjacent compartment by a magnetic member operating outside the cartridge body, the reagent supply unit has a plurality of reagent chambers formed in a row, each corresponding to the compartment, with the lower end sealed by a sealing film, and containing reagents (and washing liquid) inside, in order to supply at least reagents (or washing liquid) to each compartment in the cartridge body, a perforating pin or perforating plate is built into the inside of each compartment for piercing the sealing film when the reagent supply unit descends, and the plurality of reagent chambers can have an inclination of at least 45 degrees so that the reagent flows well into the inside of the main body after perforation.

[0014] According to the present invention, the diagnostic cartridge further includes a reagent discharge tube that connects the flow channel inlet in the reaction chamber to the cartridge body and discharges nucleic acids extracted from the cartridge body into the reaction chamber, and a syringe that is connected to the flow channel outlet in the reaction chamber and draws the extracted nucleic acids from the reagent discharge tube into the reaction chamber, and the reagent supply unit can have a sample inlet formed in front of the first reagent chamber, which corresponds to the first compartment of a plurality of compartments in the main body, corresponding to the sample supply path, for injecting a sample into the first compartment.

[0015] According to the present invention, the reagent supply unit has a nucleic acid extraction port formed at the rear end side corresponding to the final compartment of the multiple compartments of the main body for extracting nucleic acids purified from the final compartment within the main body, and nucleic acid extraction through the nucleic acid extraction port can be performed using a pipette or syringe, and the reagent discharge tube has an inverted U-shaped cross-sectional structure that connects the final compartment of the multiple compartments of the main body to the flow channel inlet of the reaction chamber in order to transport the reagent containing nucleic acids extracted from the sample to the reaction chamber, and slide grooves may be formed on both sides so as to be fitted into and fixed in the side cutouts of the main body.

[0016] According to the present invention, the reaction chamber may be fixed by fitting one end of its horizontal plane into a mounting groove formed on one side of the main body, or by locking protrusions corresponding to the locking grooves may be formed on the left and right sides of the front end so as to lock into and connect with locking grooves provided on the upper left and right sides of a stand formed on the side of the main body. In another embodiment, the reaction chamber may be formed integrally with the cartridge body.

[0017] According to the present invention, the diagnostic cartridge may be configured such that the bottom surface of the final compartment of the cartridge body and the inlet of the flow path in the reaction chamber are at the same height as each other, so that the final compartment of the cartridge body and the flow path in the reaction chamber are in communication with each other, in order to discharge nucleic acids extracted from the cartridge body into the reaction chamber. The diagnostic cartridge may further include a predetermined suction means that generates a vacuum pressure through a predetermined suction step so that nucleic acids can move from the cartridge body along the flow path in the reaction chamber, and such a suction means may consist of a cylinder integrally formed on the side surface of the cartridge body and a piston that performs the suction step along the inside of the cylinder.

[0018] According to the present invention, the main body may further include at least one ultrasonic generator provided in contact with an aluminum film attached to the lower end of the main body to provide ultrasound for smooth mixing of nucleic acid of a sample and reagent in a first compartment of a plurality of compartments of the main body, and the reaction chamber may further include at least one ultrasonic generator provided in contact with an aluminum film sealing at least a portion of the reaction chamber to provide ultrasound for unwinding double-stranded DNA into single-stranded DNA in a polymerization enzyme chain reaction process.

[0019] According to yet another embodiment of the present invention, a diagnostic cartridge for nucleic acid extraction, amplification, and analysis, comprising a cartridge body with an open top and comprising a plurality of compartments; and a plurality of chambers for containing reagents or samples formed in a row, each chamber including a reagent supply section having a 45-degree inclination on one side wall, wherein the cartridge body is provided with a puncture pin for piercing a sealing film attached to one side of the reagent supply section when the reagent supply section is attached to the open top of the cartridge body, and the plurality of compartments further comprising a reaction chamber provided on one side of the cartridge body for nucleic acid amplification, wherein at least a portion of each compartment is inclined at 45 degrees so that the reagent or sample inside the reagent supply section flows better into the cartridge body, and the plurality of compartments are configured such that a portion of the partition wall between each compartment is at a height a certain level lower than a reference height. [Effects of the Invention]

[0020] Based on the aforementioned features, the diagnostic cartridge according to the present invention can be used not only for molecular diagnosis but also for immunodiagnosis. In particular, since the nucleic acid-coated beads are transported inside the cartridge body using a magnetic component, the sample does not leak out of the cartridge, allowing for safer diagnostic work. [Brief explanation of the drawing]

[0021] [Figure 1a] Figure 1a is a perspective view showing the external appearance and components of the diagnostic cartridge of the present invention. [Figure 1b] Figure 1b is a perspective view showing the external appearance and components of the diagnostic cartridge of the present invention. [Figure 1c] Figure 1c is a perspective view showing the external appearance and components of the diagnostic cartridge of the present invention. [Figure 1d] Figure 1d is a perspective view showing the external appearance and components of the diagnostic cartridge of the present invention. [Figure 2] Figure 2 is an enlarged cross-sectional view of the reagent discharge tube and syringe attached to the cartridge body shown in Figure 1. [Figure 3a] Figure 3a is a partially broken perspective view showing the operating state of the diagnostic cartridge of the present invention. [Figure 3b] Figure 3b is a partially broken perspective view showing the operating state of the diagnostic cartridge of the present invention. [Figure 4] Figure 4 is a structural and connection diagram of the cartridge body and reaction chamber of Figure 1. [Figure 5] Figure 5 is a schematic diagram for explaining the operation of the diagnostic cartridge of the present invention. [Figure 6] Figure 6 is a perspective view showing another embodiment of the diagnostic cartridge according to the present invention. [Figure 7a] Figure 7a is a perspective view showing the shape and structure of the cartridge body provided in the diagnostic cartridge of Figure 6. [Figure 7b] Figure 7b is a perspective view showing the shape and structure of the cartridge body provided in the diagnostic cartridge of Figure 6. [Figure 8a] Figure 8a is a perspective view showing the shape and structure of the reagent supply unit provided in the diagnostic cartridge of Figure 6. [Figure 8b] Figure 8b is a perspective view showing the shape and structure of the reagent supply unit provided in the diagnostic cartridge of Figure 6. [Figure 8c] Figure 8c is a perspective view showing the shape and structure of the reagent supply unit provided in the diagnostic cartridge of Figure 6. [Figure 9a] Figure 9a is a partially broken perspective view showing the initial state of the diagnostic cartridge of Figure 6 with the reagent supply unit attached. [Figure 9b] Figure 9b is a partially broken perspective view showing the initial state of the diagnostic cartridge of Figure 6 with the reagent supply unit attached. [Figure 10a] Figure 10a is a perspective view showing the state change of the diagnostic cartridge of Figure 6 when the reagent supply unit is lowered. [Figure 10b] Figure 10b is a perspective view showing the state change of the diagnostic cartridge of Figure 6 when the reagent supply unit is lowered. [Figure 10c]Figure 10c is a perspective view showing the change in the state of the diagnostic cartridge in Figure 6 when the reagent supply unit is lowered. [Figure 11a] Figure 11a is a cross-sectional diagram illustrating the principle and pathway of reagent and nucleic acid movement within the diagnostic cartridge shown in Figure 6. [Figure 11b] Figure 11b is a cross-sectional diagram illustrating the principle and pathway of reagent and nucleic acid movement within the diagnostic cartridge shown in Figure 6. [Figure 12a] Figure 12a shows the usage conditions for extracting purified DNA from the diagnostic cartridge shown in Figure 6. [Figure 12b] Figure 12b shows the usage conditions for extracting purified DNA from the diagnostic cartridge in Figure 6. [Figure 13a] Figure 13a is a perspective view showing the shape and structure of the cartridge body of another embodiment of the diagnostic cartridge according to the present invention. [Figure 13b] Figure 13b is a perspective view showing the shape and structure of the cartridge body of another embodiment of the diagnostic cartridge according to the present invention. [Figure 14a] Figure 14a is a magnified perspective view showing the deformed internal structure of the cartridge body of the diagnostic cartridge shown in Figure 13. [Figure 14b] Figure 14b is a magnified perspective view showing the deformed internal structure of the cartridge body of the diagnostic cartridge shown in Figure 13. [Figure 15a] Figure 15a is a partially broken perspective view showing the initial state of the diagnostic cartridge in Figure 13. [Figure 15b] Figure 15b is a partially broken perspective view showing the initial state of the diagnostic cartridge in Figure 13. [Figure 16a] Figure 16a is a perspective view showing the change in the state of the diagnostic cartridge in Figure 13 when the reagent supply unit is lowered. [Figure 16b] Figure 16b is a perspective view showing the change in the state of the diagnostic cartridge in Figure 13 when the reagent supply unit is lowered. [Figure 17]Figure 17 is a cross-sectional diagram illustrating the principle and pathway of reagent and nucleic acid movement within the diagnostic cartridge shown in Figure 13. [Figure 18] Figure 18 is an illustrative diagram showing the arrangement of POCT equipment for emitting ultrasound to a reaction chamber attached to a diagnostic cartridge according to one embodiment of the present invention. [Modes for carrying out the invention]

[0022] The present invention will be described in more detail below with reference to the attached drawings and embodiments.

[0023] In the embodiments described below, illustrations and descriptions have been omitted except for those parts essential to illustrating the invention. Throughout the specification, the same reference numerals are used for similar elements, and detailed descriptions thereof are omitted without repetition.

[0024] Figures 1 and 2 show the configuration of the diagnostic cartridge for nucleic acid extraction, amplification, and analysis of the present invention. The diagnostic cartridge of the present invention consists of a cartridge body 100 (hereinafter also referred to as "body") which is a rectangular case with an open top for extracting nucleic acids from a sample, and a reaction chamber 200 which is coupled to the side of the cartridge body and performs nucleic acid amplification.

[0025] The cartridge body 100 has a reagent supply unit 300 attached through an open top, and below the reagent supply unit 300, a number of compartments 111, 112, 113, 114, 115, ... (see Figure 5) are formed in a row inside the body 100 for accommodating the supplied reagents and samples.

[0026] Multiple compartments 111, 112, 113, 114, and 115 are closed at the top by the installation of the reagent supply unit 300. Of the sequentially arranged compartments 111, 112, 113, 114, and 115, a sample supply passage 101 for introducing a sample is formed in front of the first compartment 111 and connected to the first compartment 111. The first compartment 111 contains multiple magnetic beads 102, and it is preferable that each of the compartments 111, 112, 113, 114, and 115 has a structure in which a portion of the upper part is interconnected so that the magnetic beads 102 can be transported from the first compartment 111 to the final compartment (in this embodiment, the fifth compartment 115 is shown and described as the final compartment as an example) by a predetermined magnetic member 103.

[0027] The reagent supply unit 300 has reagent chambers 311, 312, 313, 314, and 315 arranged in a row, each corresponding to one of the compartments 111, 112, 113, 114, and 115 within the main body 100, with the lower part of each chamber perforated. Each reagent chamber 311, 312, 313, 314, and 315 contains a puncturing pin (not shown) for rupturing a capsule containing the reagent (and wash solution). In the present invention, the reagent supply unit 300 can be configured such that the reagent chambers 311, 312, 313, 314, and 315 have an inclined surface of approximately 45 degrees in order to allow the reagent that has leaked out of the reagent capsule (not shown) to flow more efficiently into the main body 100. The reagent supply unit 300 has a sample inlet 301 formed in front of the first reagent chamber 311, which corresponds to the first compartment 111 of the main body 100, and which corresponds to the sample supply path 101, for injecting a sample into the first compartment 111.

[0028] The reaction chamber 200 is supplied with a nucleic acid-containing reagent from the main body 100 using the vacuum pressure of the syringe 400 to amplify the nucleic acid, and is fixed to the side of the main body 100 by being attached laterally. For fixing the reaction chamber 200, a stand 141 that supports the lower end of the reaction chamber 200 is formed on the side of the main body 100, and locking grooves 142 corresponding to the locking protrusions 220 formed on both sides of the front end of the reaction chamber 200 are formed on the upper left and right sides of the stand 141.

[0029] The reaction chamber 200 has a flow path 210 formed inside, passing through multiple chambers and probes (see Figure 4b). The end of a syringe 400 is connected to the outlet 211 of this flow path 210 (hereinafter referred to as the "flow path outlet"), and a reagent discharge tube 500 extending from the final compartment 115 inside the main body is connected to the inlet 212 of the flow path (hereinafter referred to as the "flow path inlet") (see Figure 3b).

[0030] The syringe 400 can be detachably attached to a mounting section 120 formed on the side of the main body 100, and will generate pressure to pull the fluid in order to fill the reaction chamber 200 with reagents.

[0031] The reagent discharge tube 500 is for transporting 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 final compartment 115 of the main body 100 to the flow channel inlet 212 of the reaction chamber 200, and can be provided with sliding grooves 510 on both sides so that it can be fitted into and fixed in the side cutout 130 of the main body 100.

[0032] Referring to Figure 4a, the first compartment 111 can be provided with ultrasound to better mix the nucleic acid of the sample with the reagent, and for this purpose the lower part of the first compartment 111 may be provided with a first ultrasonic generator 600 (see Figure 3a) which is provided in contact with an aluminum film 150 sealed to the lower end of the main body 100.

[0033] Furthermore, the reaction chamber 200 may be equipped with a second ultrasonic generator 700 (see Figure 3b) that can provide ultrasound to unwind double-stranded DNA into single-stranded DNA in a polymerization enzyme chain reaction process, and for this purpose is provided in contact with an aluminum film 230 sealed at the lower end of the reaction chamber 200.

[0034] The diagnostic cartridge of the present invention having the above-described configuration can be used as shown through Figure 5.

[0035] First, when the collected sample is injected through the sample inlet 301 of the reagent supply unit 300, the sample flows into the first compartment 111 through the sample supply path 101 of the cartridge body 100, rupturing a reagent capsule (not shown) from the first reagent chamber 311 of the reagent supply unit 300 and filling the first compartment 111 with reagent. In addition, multiple magnetic beads 102 are placed inside the first compartment 111, and the magnetic beads 102 are moved from the outside using a magnetic member 103 to pulverize the sample and destroy the cells, thereby ensuring that the reagent and sample are well mixed. Through this process, components leaked from the sample cells may adhere to the surface of the beads 102.

[0036] In the illustrated configuration of the present invention, wash liquid for washing can be supplied through the second reagent chamber 312, third reagent chamber 313, and fourth reagent chamber 314 of the reagent supply unit 300 by rupturing capsules, and the wash liquid flowing down from the second, third, and fourth reagent chambers 312, 313, and 314 fills the inside of the second compartment 112, third compartment 113, and fourth compartment 114, respectively.

[0037] In this state, the magnetic member 103 is used to sequentially move the magnetic beads 102 inside the first compartment 111 to the second compartment 112, the third compartment 113, and the fourth compartment 114, while cleaning to remove foreign matter from the surface of the magnetic beads 102.

[0038] Furthermore, an elution solution for nucleic acid extraction can be supplied through the fifth reagent chamber 315 of the reagent supply unit 300 by rupturing a capsule. The elution solution flowing down from the fifth reagent chamber 315 fills the inside of the fifth compartment 115, which is the final compartment. At this time, when the washed magnetic beads 102 inside the fourth compartment 114 are moved into the fifth compartment 115 using the magnetic member 103, nucleic acids are extracted from the surface of the magnetic beads 102 by the elution solution.

[0039] The extracted nucleic acids flow into the reaction chamber 200 through an U-shaped reagent discharge tube 500 by vacuum pressure created by the suction process of the syringe 400 outside the main body 100, and then undergo nucleic acid amplification as they move along the flow path 210.

[0040] The diagnostic cartridge of the present invention, which operates in this manner, has the advantage of allowing for safer diagnostic work because, since the beads to which nucleic acids are attached are transferred inside the cartridge body using a magnetic member, no sample leaks out of the cartridge.

[0041] Next, another embodiment of the diagnostic cartridge for nucleic acid extraction, amplification, and analysis of the present invention will be described with reference to Figures 6 to 13.

[0042] The diagnostic cartridge for nucleic acid extraction, amplification, and analysis of the present invention, as shown in Figure 6, comprises a cartridge body 100' (hereinafter also referred to as "body") with an open top for extracting nucleic acids from a sample, a reagent supply unit 300' inserted through the open top of the body 100' and placed on the inner upper part of the body 100', and a reaction chamber 200' coupled to the side of the body 100' for nucleic acid amplification.

[0043] Figures 7a and 7b show the shape and structure of the cartridge body illustrated in Figure 6. The body 100' is open at the top to accommodate the reagent supply unit 300', and multiple locking grooves 104a (104b) for placing the reagent supply unit 300' are formed vertically at regular intervals on the upper part of the front and rear walls. Below the reagent supply unit 300' placed in the locking grooves 104a (104b), multiple compartments 111, 112, 113, 114, 115, ... are formed in a row inside the body 100' for accommodating the supplied reagents and samples.

[0044] In this embodiment, as illustrated in Figures 7a to 7b, there are five partitions 111, 112, 113, 114, and 115, and hereafter, the fifth partition, the fifth partition 115, will be referred to as the "final partition."

[0045] Multiple compartments 111, 112, 113, 114, and 115 are closed at the top by the installation of the reagent supply unit 300'. Of the sequentially arranged compartments 111, 112, 113, 114, and 115, a sample supply passage 101 for introducing a sample is formed in front of the first compartment 111 and connected to the first compartment 111. The first compartment 111 contains multiple magnetic beads (see Figure 11a) 102. Preferably, the top of the partition wall 105 of each compartment 111, 112, 113, 114, and 115 is open so that the magnetic beads 102 can be transported from the first compartment 111 to the final compartment 115 by a predetermined magnetic member (magnet) 103, and the compartments 111, 112, 113, 114, and 115 are interconnected. Such a connecting structure can be realized, for example, by providing a recess (connecting recess) 108 in which at least a portion of the height of the partition wall 105 located between each of the partitions 111, 112, 113, 114, and 115 is formed to be lower.

[0046] Figures 8a to 8c show the shape and structure of the reagent supply unit provided in the diagnostic cartridge of Figure 6, and Figures 9a and 9b show the initial state of the diagnostic cartridge of Figure 6 with the reagent supply unit attached in the form of a partially broken perspective view. Figures 8b and 8c show that the reagent supply unit 300' has a number of reagent chambers 311, 312, 313, 314, and 315 arranged in a row inside, corresponding to the number of compartments 111, 112, 113, 114, and 115 in the main body 100', for supplying reagents (or wash liquid) to each compartment 111, 112, 113, 114, and 115.

[0047] Such reagent chambers 311, 312, 313, 314, and 315 are formed diagonally downward and rearward from inside the reagent supply section 300', with an open bottom and an upper end closed by the upper surface of the reagent supply section 300'. Each reagent chamber 311, 312, 313, 314, and 315 may be provided filled with a reagent (or wash solution) and with its open bottom sealed by a predetermined film material (sealing film, see Figure 9b) 310a.

[0048] On the other hand, in order to pierce through such a sealing film 310a and supply the reagent (or wash solution) inside to the interior of each compartment 111, 112, 113, 114, and 115 of the main body 100', the cartridge body 100' has perforating pins 107 (see Figure 7b) that protrude upward in a pointed manner, corresponding to the sealing film 310a of each reagent chamber 311, 312, 113, 114, and 115.

[0049] Referring to Figures 8b and 9b, the reagent chambers 311, 312, 313, 314, and 315, which are filled with reagents (or wash liquid), have a structure that extends diagonally downward from a closed top surface to the rear, and as mentioned above, a sealing film 310a is attached to the opening at the lower end to prevent the reagents (or wash liquid) from flowing out. As shown in the figures, each reagent chamber 311, 312, 313, 314, and 315 is preferably inclined at approximately 45 degrees toward the front of the main body 100' inside the reagent supply section 300', and with the reagent chambers 311, 312, 313, 314, and 315 being inclined at approximately 45 degrees, the lower ends of each reagent chamber 311, 312, 313, 314, and 315 sealed by the sealing film 310a are also inclined at approximately 45 degrees from the horizontal (see Figure 8c). Because the reagent chambers 311, 312, 313, 314, and 315 each have a 45-degree inclination angle, when the sealing film 310a is perforated, the reagent will flow in a single direction along the inclined surface, thus preventing any residual reagent from remaining in the chamber (see Figure 8c).

[0050] Furthermore, it is preferable that at least a portion of the rear wall surface 106 of each of the compartments 111, 112, 113, 114, and 115 of the main body 100' have an inclination angle of about 45 degrees so that the reagents supplied from the reagent chambers 311, 312, 313, 314, and 315 at an inclination of about 45 degrees can flow down smoothly.

[0051] In this embodiment, Figure 8a shows a perspective view of the reagent supply unit from the front, and Figure 8b shows a perspective view of the reagent supply unit from the rear. At one end of the reagent supply unit 300', preferably in the forward direction of the first reagent chamber 311 among the reagent chambers 311, 312, 313, 314, and 315, a sample inlet 301 for injecting a sample into the first compartment 111 within the main body 100' is formed.

[0052] The sample inlet 301 extends downward so as to be close to the sample supply passage 101 of the main body 100', and the inlet of the sample inlet 301 may be provided with an open / close door 302 that can be opened and closed for sample injection. The open / close door 302 may be configured to slide along guides 303 that are positioned at a certain distance apart on the upper front end surface of the reagent supply section 300', and for this purpose, guide grooves 302a are formed on both sides of the open / close door 302 for fitting and attaching to the guides 303.

[0053] Furthermore, on the upper surface of the reagent supply section 300' in another direction, preferably on the same line as the fifth reagent chamber 315 (in this embodiment, the fifth reagent chamber 315 is shown and described as the "final reagent chamber") among the reagent chambers 311, 312, 313, 314, and 315, a nucleic acid extraction port 305 for extracting nucleic acids from inside the final compartment 115 in the main body 100' extends downward through the inside of the reagent supply section 300', corresponding to the final compartment 115.

[0054] The entrance to the nucleic acid extraction port 305 may be provided with an openable / closable door 306 for nucleic acid extraction, and such an openable / closable door 306 may be configured to slide along guides 307 that are positioned at a certain distance apart on the upper surface of the final reagent chamber 315. For this purpose, guide grooves 306a are formed on both sides of the openable / closable door 306 for fitting and attaching to the guides 307.

[0055] The aforementioned opening and closing doors 302 (306) may be provided with a handle protruding from the top surface to facilitate opening and closing by the user. Preferably, stopper projections 303a (307a) for each opening and closing door 302 (306) are formed at the front end of the guide 303 (307) on which the opening and closing doors 302 (306) slide, i.e., at the sample inlet 301 side and the nucleic acid extraction port 305 side, and fitting grooves 302b (306b) are formed at the front end of the opening and closing doors 302 (306) so as to fit into and secure them to the stopper projections 303a (307a) respectively (see Figures 8b and 8c).

[0056] On the front and rear surfaces of the reagent supply section 300', locking projections 308 are provided for securing the reagent supply section 300' in the locking grooves 104a (104b) of the main body 100'. The part labeled "304" not shown in the drawing indicates a space created by removing material from the reagent supply section 300'.

[0057] The reaction chamber 200' is supplied with a reagent containing nucleic acid from the main body 100 using a vacuum pressure via a syringe 400 to amplify the nucleic acid. In this embodiment, the description will focus on a binding structure as shown in Figure 6, in which the reaction chamber 200' is horizontally attached to the side of the main body 100'.

[0058] For mounting the reaction chamber 200', a mounting groove 142 is formed on the side of the main body 100', preferably the side of the main body 100' in the direction of the final partition chamber 115, for fitting and securing one end of the horizontal plane of the reaction chamber 200'.

[0059] In the following description, the reaction chamber 200' is described as having a structure in which one end of its horizontal plane is fitted into a mounting groove 142 formed on the side of the main body 100' and fixed in place. However, it is not limited to this, and as shown in Figure 1b of the above embodiment, locking protrusions 220 are formed on both sides of the front end of the reaction chamber 200, and locking grooves 142 are formed on the upper left and right sides of the mounting base 141 formed on the side of the main body 100, and when the reaction chamber 200 is joined, the locking protrusions 220 are locked into the locking grooves 142 to join it.

[0060] The internal structure of the reaction chamber 200' may be provided in the same form as that described in the above embodiment. That is, as shown in Figure 4b, a flow path 210 passing through multiple chambers and probes is formed inside the reaction chamber 200, the end of a syringe 400 is connected to the outlet 211 of the flow path 210 (hereinafter referred to as the "flow path outlet"), and a reagent discharge tube 500 extending from the final compartment 115 in the main body 100' is connected to the inlet 212 of the flow path (hereinafter referred to as the "flow path inlet") (see Figure 5).

[0061] In this embodiment, the syringe 400 can be detachably attached to a stand 120 formed on the side of the main body 100', thereby creating a pressure that pulls the fluid to fill the reaction chamber 200' with reagents, i.e., a negative suction pressure.

[0062] In this embodiment, the diagnostic cartridge further includes a reagent discharge tube 500 that connects the final compartment 115 of the main body 100 to the reaction chamber 200'. The reagent discharge tube 500 is for transporting the reagent containing nucleic acid extracted from the sample to the reaction chamber 200', and has a discharge passage 501 with an inverted U-shaped cross-sectional structure that connects the final compartment 115 of the main body 100 to the flow path inlet 212 of the reaction chamber 200', and has a structure in which slide grooves 510 are formed on both sides so that it can be fitted into and fixed in the side cutout 130 of the main body 100.

[0063] Figures 10a to 10c show the change in the state of the diagnostic cartridge when the reagent supply unit 300' shown in Figure 9, which is attached to the top of the main body, is pushed downward. Referring to Figures 9a and 9b, in the initial state, the reagent supply unit 300' is located on the upper inside of the main body 100'. In this state, the locking projections 308 protruding from the front and rear surfaces of the reagent supply unit 300' are locked into the upper locking grooves 104a formed on the front and rear surfaces of the main body 100', supporting the reagent supply unit 300'. At this time, it can be seen that the puncture pins 107 formed in each of the compartments 111, 112, 113, 114, and 115 of the main body 100' maintain a state where they are spaced at a constant distance downward from the sealing film 310a attached to the lower end surfaces of the reagent chambers 311, 312, 313, 314, and 315.

[0064] Subsequently, when the reagent supply unit 300' is pushed downward, causing the reagent chambers 311, 312, 313, 314, and 315 to descend as shown in Figures 10a and 10b, the locking projections 308 protruding from the front and rear surfaces of the reagent supply unit 300' also descend downward and are placed in a locked state in the lower locking grooves 104b formed on the front and rear surfaces of the main body 100', thereby supporting the reagent supply unit 300'.

[0065] At this time, as the reagent chambers 311, 312, 313, 314, and 315 descend, the sealing film 310a covering the lower end surfaces of the reagent chambers 311, 312, 313, 314, and 315 is pierced by the perforating pins 107 formed in each of the compartments 111, 112, 113, 114, and 115 of the main body 100', as shown in Figure 10c. As a result, the reagent (or wash solution) filled in the reagent chambers 311, 312, 313, 314, and 315 flows along the rear wall surface 106 inside the main body 100' and is supplied into each of the compartments 111, 112, 113, 114, and 115.

[0066] The diagnostic cartridge of the present invention having the above-described configuration can be used as shown through Figures 11a and 11b.

[0067] Figures 11a and 11b are cross-sectional diagrams illustrating the principle and pathway of reagent and nucleic acid movement within the diagnostic cartridge of Figure 6. First, a sample is taken from the human body, and the taken sample is injected through the sample inlet 301 of the reagent supply unit 300'. The sample then flows into the first compartment 111 through the sample supply path 101 of the main body 100'.

[0068] At this time, when the reagent supply unit 300' attached to the upper inner part of the main body 100' is pushed downward, the sealing film 310a that covers the lower end surfaces of the reagent chambers 311, 312, 313, 314, and 315 is punctured by the perforating pins 107 in each of the compartments 111, 112, 113, 114, and 115 inside the main body 100' (see Figure 10c), thereby causing the reagent (or wash liquid) to flow down from each of the reagent chambers 311, 312, 313, 314, and 315 and fill the inside of each of the compartments 111, 112, 113, 114, and 115.

[0069] Multiple magnetic beads 102 are placed inside the first compartment 111, and the magnetic beads 102 are moved from the outside using a magnetic component (magnet) 103 to pulverize the sample and destroy the cells, thereby ensuring that the reagent and the sample are thoroughly mixed. Through this process, components leaked from the sample cells may adhere to the surface of the beads 102.

[0070] In this embodiment, the second, third, and fourth reagent chambers 312, 313, and 314 of the reagent supply unit 300' contain a wash solution for cleaning. As the reagent supply unit 300' descends, the sealing film 310a is perforated by the perforating pin 107, and the wash solution in the second, third, and fourth reagent chambers 312, 313, and 314 fills the inside of the second, third, and fourth compartments 112, 113, and 114. In this state, the magnetic member 103 is used to sequentially move the magnetic beads 102 inside the first compartment 111 to the second, third, and fourth compartments 112, 113, and 114, removing foreign matter from the surface of the magnetic beads 102 through a cleaning process with the wash solution.

[0071] In this embodiment, the final reagent chamber 315 of the reagent supply unit 300' contains an elution solution for nucleic acid extraction. As the reagent supply unit 300' descends, the sealing film 310a is perforated by the perforating pin 107, causing the elution solution in the final reagent chamber 315 to fill the fifth chamber 115, which is the final chamber in the main body 100'. At this time, the washed magnetic beads 102 inside the fourth chamber 114 are moved into the final chamber 115 using the magnetic member 103, and nucleic acids (DNA) are extracted from the surface of the magnetic beads 102 by the elution solution.

[0072] At this time, the extracted nucleic acid (DNA) is transferred to the upper part of the reagent level above the precipitated magnetic beads 102. However, it can be discharged into the reaction chamber 200' through the U-shaped discharge channel 501 in the reagent discharge tube 500 by the vacuum pressure created by the suction process of the syringe 400 attached to the side of the main body 100. After the nucleic acid (DNA) flows into the reaction chamber 200', nucleic acid amplification is performed as it moves along the flow path 210.

[0073] Generally, various factors affect the amplification efficiency of PCR. Common factors include time, temperature, amount and properties of polymerase, amount of DNA, 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 are holding it from the extraction stage. Therefore, in this embodiment, a magnet 103 is used to hold the beads 102.

[0074] Furthermore, when the purified DNA is sent to the reaction chamber 200', residual reagent that does not adhere to the magnet may flow into the reaction chamber 200'. In this embodiment, to prevent this, it is preferable that the inlet of the U-shaped discharge channel 501 in the reagent discharge tube 500 be located at a water level higher than the beads 102 so that the DNA is drawn in at a water level higher than the beads 102.

[0075] The diagnostic cartridge of the present invention, having the configuration and operation described above, can be used not only for molecular diagnosis but also for immunodiagnosis. In particular, since magnetic beads to which nucleic acids are attached are transported inside the cartridge body using a magnet, the sample does not leak out of the cartridge, thus offering the advantage of safer diagnostic work.

[0076] Figures 12a and 12b illustrate an example of use for extracting purified DNA within the diagnostic cartridge of the present invention, where a pipette (P, or syringe) can be used to extract the purified DNA from the fifth chamber 115, which is the final chamber.

[0077] Generally, conventional POCT (Point of Care Testing) cartridges are used for only one test, meaning the purified DNA obtained through the process within the cartridge cannot be removed. Therefore, if a test fails with a POCT setup, the purified DNA in the cartridge cannot be used again, necessitating the acquisition of a new sample.

[0078] However, according to the present invention, purified DNA can be extracted by opening the opening / closing door 306 and inserting the pipette P into the final compartment 115 through the nucleic acid extraction port 305.

[0079] As a result, even if the POCT (Point-of-Cognitive Testing) test fails, the extracted DNA is from the same patient, allowing for retesting without the need to repeatedly obtain samples as in conventional methods. Of course, the extracted DNA can also be used for other purposes.

[0080] Figures 13a and 13b show yet another embodiment of the diagnostic cartridge according to the present invention, in which the reaction chamber 200” does not require a configuration such as the stand 141 and locking groove 142 shown in the configurations of Figures 1 and 6, and may be integrally formed with the cartridge body 100” so that the flow path 210 inside it and the final compartment 115' of the cartridge body are in communication with each other, and the syringe 400” can be provided with a structure in which the surface of the cylinder 410 is integrally formed with the cartridge body 100” with its end integrally coupled to the flow path outlet 211 of the reaction chamber 200”.

[0081] In particular, in this embodiment, the diagnostic cartridge can be configured by removing the reagent discharge tube 500 from the configuration in Figures 1 and 6, and by increasing the height of the bottom of the final compartment 115' of the cartridge body 100" so that it is the same height as the reaction chamber 200". This allows the final compartment 115' and the flow path 210 of the reaction chamber 200" to communicate with each other even without the reagent discharge tube 500 (Figures 1 and 6).

[0082] Figures 14a and 14b show enlarged views of the deformed internal structure of the cartridge body provided in the diagnostic cartridge of Figure 13. In the configuration of Figure 7b, the perforating pins 107, which protrude upward in a pointed shape corresponding to the sealing films 310a of each reagent chamber 311, 312, 313, 314, and 315, may be composed of panel-shaped perforating plates 107' in this embodiment. Such perforating plates 107' preferably consist of at least one pair of panels arranged side by side at regular intervals from each other.

[0083] Figures 15 and 16 are partially broken perspective views showing the change in the state of the diagnostic cartridge before and after the descent of the reagent supply unit. In the initial state shown in Figure 15, the reagent supply unit 300' is located on the upper inner side of the main body 100'. In this state, the locking protrusions 308' protruding from the left and right surfaces of the reagent supply unit 300' are locked onto the locking protrusions 104c formed on the left and right surfaces of the main body 100', supporting the reagent supply unit 300'. At this time, it can be seen that the perforated plates 107' formed in each of the compartments 111, 112, 113, 114, and 115' of the main body 100' maintain a state where they are spaced at regular intervals downward from the sealing film 310a attached to the lower end surfaces of the reagent chambers 311, 312, 313, 314, and 315.

[0084] Thereafter, by pushing the reagent supply unit 300' downwards and lowering the reagent chambers 311, 312, 313, 314, and 315 as shown in Figure 16a, the sealing film 310a that covers the lower end surfaces of the reagent chambers 311, 312, 313, 314, and 315 is punctured by the perforated plates 107' formed in each of the compartments 111, 112, 113, 114, and 115' of the cartridge body 100'' as shown in Figure 16b. As a result, the reagent (or wash liquid) filled in the reagent chambers 311, 312, 313, 314, and 315 flows along the rear wall surface 106 inside the body 100'' and can be supplied into each of the compartments 111, 112, 113, 114, and 115'.

[0085] Figure 17 is a cross-sectional diagram illustrating the principle and pathway of reagent and nucleic acid movement within the diagnostic cartridge shown in Figure 13. After a sample taken from the human body is introduced into the first compartment 111 through the sample inlet 301 of the reagent supply unit 300', as shown in Figure 16, the reagent supply unit 300' is pushed downward to fill the compartments 111, 112, 113, 114, and 115' with reagents (or wash liquid) from each reagent chamber 311, 312, 313, 314, and 315'. Then, multiple magnetic beads 102 are introduced into the first compartment 111, and the magnetic beads 102 are moved from the outside using a magnetic member (magnet) 103 to pulverize the sample and destroy the cells, thereby ensuring that the reagent and sample are well mixed. Through this process, components leaked from the cells of the sample may adhere to the surface of the beads 102.

[0086] In this embodiment, the second, third, and fourth reagent chambers 312, 313, and 314 of the reagent supply unit 300' contain a wash liquid for cleaning. As the reagent supply unit 300' descends, the sealing film 310a is perforated by the perforating plate 107', and the wash liquid in the second, third, and fourth reagent chambers 312, 313, and 314 fills the inside of the second, third, and fourth compartments 112, 113, and 114. In this state, the magnetic member 103 is used to sequentially move the magnetic beads 102 inside the first compartment 111 to the second, third, and fourth compartments 112, 113, and 114, removing foreign matter from the surface of the magnetic beads 102 through a cleaning process with the wash liquid.

[0087] In this embodiment, the final reagent chamber 315 of the reagent supply unit 300' contains an elution solution for nucleic acid extraction. As the reagent supply unit 300' descends, the sealing film 310a is perforated by the perforating plate 107', causing the elution solution in the final reagent chamber 315 to fill the fifth chamber 115', which is the final chamber in the main body 100'. At this time, the washed magnetic beads 102 inside the fourth chamber 114 are moved into the final chamber 115' using the magnetic member 103, and nucleic acids (DNA) are extracted from the surface of the magnetic beads 102 by the elution solution.

[0088] At this time, the extracted nucleic acid (DNA) can be discharged into the reaction chamber 200' by the vacuum pressure created by the suction process of the syringe 400 which is integrally attached to the flow channel outlet 211 of the reaction chamber. After flowing into the reaction chamber 200', the nucleic acid (DNA) moves along the flow channel 210 while nucleic acid amplification takes place.

[0089] Figure 18 shows the configuration, arrangement, and usage of a POCT (Point of Use Test) inspection device for emitting ultrasound to a reaction chamber attached to a diagnostic cartridge according to one embodiment of the present invention. The reaction chamber 200' can be supplied with ultrasound to unwind double-stranded DNA into single-stranded DNA in a polymerization enzyme chain reaction process. For this purpose, the reaction section within the reaction chamber 200', which contains the DNA-containing reagent, is sealed with a metal film, such as an aluminum film 230, and an ultrasonic generator 700' is placed in contact with the aluminum film 230.

[0090] Such ultrasonic generators 700' raise the temperature to 95°C to denaturate and inactivate most of the DNA, but if the temperature of the denaturation and inactivation interval can be lowered or eliminated during the DNA amplification process, a considerable amount of time can be saved in existing PCR amplification processes where temperature cycles are repeated.

[0091] Although various embodiments of the present invention have been described above, the content described herein is merely an example of some preferred embodiments of the present invention and is not limited by the content described above, except as can be shown in the appended claims below. Therefore, a person with ordinary skill in the same art should understand that many variations, modifications, and substitutions of equivalents can be made within the scope of the following claims without deviating from the technical idea and essence of the invention.

Claims

1. A diagnostic cartridge for nucleic acid extraction, amplification, and analysis, A cartridge body having multiple compartments arranged in a row inside to accommodate supplied reagents and samples, the top of each compartment being open to allow reagent supply, and a sample supply path for introducing a sample into the first compartment being formed in front of the first compartment. A reagent supply unit containing at least a reagent (or wash solution), which is inserted and placed through the open top of the cartridge body, and A diagnostic cartridge comprising a reaction chamber for nucleic acid amplification, provided on one side of the cartridge body.

2. The diagnostic cartridge according to claim 1, characterized in that each partition wall between a plurality of partitions of the cartridge body is formed to be lower in height than the surrounding area so that nucleic acids attached to magnetic beads in the partition can be transported to the adjacent partition by a magnetic member operating outside the cartridge body.

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

4. The diagnostic cartridge according to claim 1, characterized in that one or more puncturing pins or puncturing plates are built into each compartment of the cartridge body for piercing the sealing film at the lower end of the reagent supply unit when the reagent supply unit descends.

5. The aforementioned diagnostic cartridge is In order to discharge the nucleic acid extracted from the cartridge body into the reaction chamber, the bottom surface of the final compartment of the cartridge body and the inlet of the flow path in the reaction chamber are formed at the same height so that they are in communication with each other. The diagnostic cartridge according to claim 1, further comprising a predetermined suction means for generating a vacuum pressure through a predetermined suction step so that nucleic acids can move from the cartridge body along the flow path in the reaction chamber.

6. The diagnostic cartridge according to claim 1, wherein the reagent supply unit is configured such that, in order to supply at least one reagent (or wash solution) to each compartment in the cartridge body, a plurality of reagent chambers are formed in a row inside the reagent supply unit, with the reagent (and wash solution) contained within the reagent supply unit, and the lower ends of each compartment in the cartridge body are sealed with a sealing film, and the lower ends of the plurality of reagent chambers are inclined at least 45 degrees.

7. The diagnostic cartridge according to claim 6, characterized in that the reagent supply unit has a sample inlet formed in front of the first reagent chamber, which corresponds to the first compartment among a plurality of compartments of the cartridge body, corresponding to the sample supply path, for injecting a sample into the first compartment.

8. The reagent supply unit has a nucleic acid extraction unit formed at the rear end side corresponding to the final compartment of the multiple compartments of the main body for extracting nucleic acids purified from the final compartment within the main body, and the nucleic acid extraction through the nucleic acid extraction unit is performed using a pipette or syringe, as described in claim 6.

9. The diagnostic cartridge according to claim 1, further comprising at least one ultrasonic generator provided in contact with an aluminum film attached to the lower end of the cartridge body, so as to provide ultrasound in a first chamber of the cartridge body for smooth mixing of nucleic acid of a sample and a reagent.

10. The diagnostic cartridge according to claim 1, further comprising at least one ultrasonic generator provided in contact with an aluminum film sealing at least a portion of the reaction chamber to provide ultrasound for unwinding double-stranded DNA into single-stranded DNA in a polymerization enzyme chain reaction process.

11. A diagnostic cartridge for nucleic acid extraction, amplification, and analysis, A cartridge body with an open top for extracting nucleic acids from a sample. The reagent supply unit is inserted and placed through the open upper part of the cartridge body. A reaction chamber for nucleic acid amplification is integrally formed on the side of the cartridge body. The extracted nucleic acid is connected to move from the cartridge body to the reaction chamber. A diagnostic cartridge comprising a suction means connected to a flow path outlet in the reaction chamber, which fills the reaction chamber with nucleic acids from the cartridge body using vacuum pressure from a suction step.

12. The aforementioned multiple partitions are, The diagnostic cartridge according to claim 11, characterized in that a portion of the partition wall between the plurality of compartments is configured to be at a height a certain level lower than the reference height.

13. The aforementioned suction means is The diagnostic cartridge according to claim 11, characterized in that it comprises a cylinder integrally formed with the cartridge body and a separate piston.

14. The diagnostic cartridge according to claim 11, characterized in that the cartridge body further includes a pipette or syringe as an extrusion means.

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