Diagnostic cartridge for extraction, amplification and analysis of nucleic acids

Abstract

The present invention relates to a diagnostic cartridge for the extraction, amplification and analysis of nucleic acids. The diagnostic cartridge comprises: a cartridge body in which a plurality of compartments having open upper portions are formed inside in a row to accommodate supplied reagents and samples, a sample supply path for introducing a sample is formed in front of a first compartment among the plurality of compartments and connected to the first compartment, and the first compartment is configured to accommodate a plurality of magnetic beads inside and transport the plurality of magnetic beads from the first compartment up to the last compartment by means of a magnetic member from the outside, wherein each partition wall between the plurality of compartments forms a structure in which the compartments communicate with each other to allow the transport of the magnetic beads; a reagent supply unit inserted and seated through the open upper portion of the cartridge body; and a reaction chamber which is coupled to the side surface of the cartridge body and in which nucleic acid amplification is performed.

Description

Technical Field

[0001] The present disclosure relates to a diagnostic cartridge for extracting high-purity nucleic acids from high-viscosity biological samples such as sputum and whole blood and, more particularly, to a diagnostic cartridge for extraction, amplification, and analysis of nucleic acids.Background Art

[0002] In modern times, with the advancement of biotechnology, interpreting the causes of diseases becomes possible at the genetic level. Accordingly, there is a growing demand for manipulation and biochemical analysis of biological samples in order to cure or prevent human diseases.

[0003] In general, diagnostic analysis tests for the biological samples refer to tests for determining the presence or absence of disease infection by detecting or measuring specific indicator substances from the biological samples such as blood, urine, and saliva. Such diagnostic analysis tests for biological samples are widely used not only for diagnostic purposes such as etiology identification, treatment, prevention, and prognosis of diseases, but also in various fields such as customized new drug development, forensic medicine, and environmental hormone detection.

[0004] As a well-known method of a diagnostic analysis test for biological samples, there is a Polymerase Chain Reaction (PCR) test that amplifies biological genes by using bacterial DNA so as to confirm the presence or absence of bacterial infection.

[0005] In order to conduct the Polymerase Chain Reaction (PCR) test, a preprocessing stage for extracting and amplifying nucleic acids, which are genetic materials such as DNA and RNA, from the biological samples is prerequisite.

[0006] In addition, other than the disease diagnosis, technology for extracting and analyzing the nucleic acids from the biological samples or cell-containing samples is required in various fields such as new drug development, preliminary testing on the presence of viral or bacterial infection, and forensic medicine.

[0007] Conventional equipment for nucleic acid extraction requires separate devices for each processing stage (e.g., concentration, purification, etc.), and accordingly, due to the migration from one device to another, it takes a long time for the processing to resume after completing a certain processing stage.

[0008] In addition, there are requirements to develop a diagnostic cartridge capable of performing diagnostic analysis tests more safely due to a sample that may leak outside the cartridge.

[0009] Meanwhile, US Patent No. 6,374,684 was proposed as a method to solve the problem of low detection efficiency due to the long processing stages of the conventional equipment for the nucleic acid extraction. In this patent, a plurality of branched flow paths is created in a piston head so as to directly intake reagents contained in a chamber while the piston head rotates, thereby mixing them in the internal space of the piston.

[0010] However, in the case of U.S. Patent No. 6,374,684, there is a problem in that the manufacturing cost of the piston may increase due to the plurality of branched flow paths that has to be created inside the piston head, so it is required to develop a cheaper and simpler cartridge structure for the nucleic acid extraction.Disclosure Technical Problem

[0011] Accordingly, the embodiment of the present disclosure is developed to solve such a problem, and an objective of the present disclosure is to provide a diagnostic cartridge for extraction, amplification, and analysis of nucleic acids, the diagnostic cartridge being able to not only perform the diagnosis safely without leakage of the sample outside the cartridge, but also be provided easily at a lower cost.Technical Solution

[0012] According to one aspect of the present disclosure to achieve the above-described objective, there is provided a diagnostic cartridge for extraction, amplification, and analysis of nucleic acids, the diagnostic cartridge including: a cartridge body configured to have a plurality of compartments formed in a row therein, each having an open upper part thereof, to accommodate a supplied reagent and a supplied sample, and have a sample supply path formed in front of a first compartment among the plurality of compartments to introduce the sample and connected to the first compartment, wherein the first compartment is configured to contain a plurality of magnetic beads so as to allow a magnetic member to transport, from outside, the plurality of magnetic beads from the first compartment to a rearmost compartment, and each partition wall between the plurality of compartments has a structure allowing each partition wall to communicate one another in order to transport the magnetic beads; a reagent supply unit configured to contain at least the reagent (or a wash solution) and to be inserted and seated through the open upper part of the cartridge body; and a reaction chamber provided on one side of the cartridge body to perform the nucleic acid amplification.

[0013] According to the present disclosure, each partition wall between the plurality of compartments may have a part thereof formed at a height lower than surrounding components, so as to allow the magnetic member operating outside the cartridge body to transport the nucleic acids adhered to the magnetic beads within one compartment to another adjacent compartment. The reagent supply unit may have a plurality of reagent chambers formed in a row and each provided with a lower end thereof sealed with a sealing film corresponding to each compartment in a state of accommodating the reagent (and the wash solution) therein in order to supply at least the reagent (or the wash solution) to each compartment within the cartridge body, perforation pins or perforation plates may be built in an interior of each compartment in order to perforate the sealing film when the reagent supply unit is lowered, and the plurality of reagent chambers may have an incline of at least 45 degrees so as to allow the reagent to flow better into an interior of the cartridge body after the perforation.

[0014] According to the present disclosure, the diagnostic cartridge may further include: a reagent discharge pipe configured to connect an inlet of a flow path within the reaction chamber to the cartridge body and discharge a nucleic acid extracted from the cartridge body into the reaction chamber; and a syringe connected to an outlet of the flow path within the reaction chamber and configured to suction the extracted nucleic acid from the reagent discharge pipe into the reaction chamber. In the reagent supply unit, a sample injection port for injecting the sample into the first compartment may be formed, corresponding to the sample supply path, at a front of a first reagent chamber corresponding to the first compartment among the plurality of compartments of the main body.

[0015] According to the present disclosure, in the reagent supply unit, a nucleic acid extraction unit for extracting a purified nucleic acid from the rearmost compartment within a main body may be formed on a rear end side corresponding to the rearmost compartment among the plurality of compartments of the main body, and the nucleic acid extraction through the nucleic acid extraction unit may use a pipette or a syringe, and the reagent discharge pipe may have a ∩-shaped cross-sectional structure connecting the rearmost compartment among the plurality of compartments of the main body and the inlet of the flow path of the reaction chamber to each other in order to transport the reagent containing the nucleic acid extracted from the sample to the reaction chamber and may have slide grooves formed on opposite sides thereof so as to be fitted and fixed into a side cutout part of the main body.

[0016] According to the present disclosure, the reaction chamber may be formed with hooking projections corresponding to hooking grooves on left and right sides of a front end thereof, so as to be fixed by one end of a horizontal surface thereof fitted into a mounting groove formed on one side of a side of the main body or be coupled to and fitted into the hooking grooves provided on upper left and right sides of a stand formed on the side of the main body. In another form, the reaction chamber may also be formed integrally with the cartridge body.

[0017] According to the present disclosure, in the diagnostic cartridge, a bottom surface of the rearmost compartment and the inlet of the flow path in the reaction chamber may be formed at the same height so as to allow the rearmost compartment of the cartridge body to communicate with the flow path in the reaction chamber in order to discharge the nucleic acid extracted from the cartridge body into the reaction chamber. The diagnostic cartridge may further include a designated suction means for generating vacuum pressure according to a predetermined suction stroke so as to enable the nucleic acids to move along the flow path within the reaction chamber from the cartridge body. Such a suction means may be configured to include: a cylinder integrally formed on a side of the cartridge body; and a piston for performing the suction stroke along an interior of the cylinder.

[0018] According to the present disclosure, the diagnostic cartridge may further include at least one ultrasonic generator provided in contact with an aluminum film attached to a lower end part of the main body, so as to provide ultrasonic waves for smooth mixing of the nucleic acids of the sample and the reagent in the first compartment among the plurality of compartments of the cartridge body, and the reaction chamber may further include at least one ultrasonic generator provided in contact with an aluminum film sealing at least a part of the reaction chamber, so as to provide ultrasonic waves in order to unwind double-stranded DNA into single strands in a polymerase chain reaction process.

[0019] According to another aspect of the present disclosure, there is provided a diagnostic cartridge for extraction, amplification, and analysis of nucleic acids, the diagnostic cartridge including: a cartridge body having an open upper part thereof and configured with a plurality of compartments; and a reagent supply part having a plurality of chambers formed in a row for accommodating a reagent or a sample, each chamber having a side wall thereof inclined at a 45-degree angle, wherein the cartridge body has a perforation pin formed in each of the plurality of compartments for perforating a sealing film attached to one side of the reagent supply part when the reagent supply part is mounted on the open upper part of the cartridge body, and the plurality of compartments has at least a part of each compartment inclined at the 45 degree angle so that the reagent or sample inside the reagent supply part flows better into an interior of the cartridge body. The diagnostic cartridge may further include a reaction chamber provided on one side of the cartridge body to perform nucleic acid amplification, and the plurality of compartments may be configured such that a part of each partition wall between each compartment has a height that is a predetermined level lower than a reference height.Advantageous Effects

[0020] From the features described above, the diagnostic cartridge according to the present disclosure may be used for not only molecular diagnosis but also immunological diagnosis, and in particular, since the magnetic beads adhered with the nucleic acids are transported by using a magnetic member inside the cartridge body, the diagnostic cartridge may perform more safe diagnostic work due to the sample not leaking out of the cartridge.Description of Drawings

[0021] FIGS. 1a to 1d are perspective views illustrating the appearance and configuration of each unit and part of a diagnostic cartridge of the present disclosure. FIG. 2 is enlarged cross-sectional views of a reagent discharge pipe and a syringe, both of which are combined with a cartridge body of FIG. 1. FIGS. 3a and 3b are partial cutaway perspective views illustrating the operating states of the diagnostic cartridge of the present disclosure. FIG. 4 are views illustrating a configuration and a connection structure of the cartridge body and reaction chamber of FIG. 1. FIG. 5 is a schematic view illustrating the operation of the diagnostic cartridge of the present disclosure. FIG. 6 is a perspective view illustrating another exemplary embodiment of a diagnostic cartridge according to the present disclosure. FIGS. 7a and 7b are perspective views illustrating the shape and structure of a cartridge body provided in the diagnostic cartridge of FIG. 6. FIGS. 8a and 8c are perspective views illustrating the shape and structure of a reagent supply unit provided in the diagnostic cartridge of FIG. 6. FIGS. 9a and 9b are partial cutaway perspective views illustrating the initial state of the diagnostic cartridge of FIG. 6 mounted with the reagent supply unit. FIGS. 10a to 10c are perspective views illustrating the change in state of the diagnostic cartridge of FIG. 6 when the reagent supply unit is lowered. FIGS. 11a and 11b are cross-sectional example diagrams illustrating the movement principle and path of a reagent and nucleic acids in the diagnostic cartridge of FIG. 6. FIGS. 12a and 12b are views illustrating a use state for extracting the purified DNA within the diagnostic cartridge of FIG. 6. FIGS. 13a and 13b are perspective views illustrating the shape and structure of a cartridge body as yet another exemplary embodiment of a diagnostic cartridge according to the present disclosure. FIGS. 14a and 14b are enlarged perspective views illustrating a modified internal structure of the cartridge body provided in the diagnostic cartridge of FIG. 13. FIGS. 15a and 15b are partial cutaway perspective views illustrating the initial state of the diagnostic cartridge of FIG. 13. FIGS. 16a and 16b are perspective views illustrating the change in state of the diagnostic cartridge of FIG. 13 when a reagent supply unit is lowered. FIG. 17 is a cross-sectional example diagram illustrating the movement principle and path of a reagent and nucleic acids in the diagnostic cartridge of FIG. 13. FIG. 18 is an example diagram illustrating the arrangement of POCT equipment for emitting ultrasonic waves to a reaction chamber mounted on a diagnostic cartridge according to one exemplary embodiment of the present disclosure. Mode for Invention

[0022] The present disclosure is described in more detail through the attached drawings and exemplary embodiments.

[0023] In the exemplary embodiments below, the illustrations and descriptions are omitted except for the parts that are essential for describing the present disclosure, and the same reference numerals are given to the same and similar components throughout the present specification, and detailed descriptions thereof are omitted without repetition.

[0024] FIGS. 1 and 2 illustrate the configuration of a diagnostic cartridge for extraction, amplification, and analysis of nucleic acids of the present disclosure. The diagnostic cartridge of the present disclosure is configured to include: a cartridge body 100 (also referred to as a "main body" hereinafter) which is a rectangular casing with an open upper part for extracting the nucleic acids from a sample; and a reaction chamber 200 coupled to a side of the cartridge body and for performing the nucleic acid amplification.

[0025] The cartridge body 100 has a reagent supply unit 300 mounted through an open upper part thereof, and a plurality of compartments 111, 112, 113, 114, 115,..., etc., (see FIG. 5) for accommodating the supplied reagent and sample is formed in a row inside the main body 100 below the reagent supply unit 300.

[0026] The upper parts of the plurality of compartments 111, 112, 113, 114, are 115 are closed by the mounted reagent supply unit 300. A sample supply path 101 for introducing the sample is formed at a front of a first compartment 111 among the sequentially arranged compartments 111, 112, 113, 114, and 115 and is connected to the first compartment 111. The first compartment 111 has a plurality of magnetic beads 102 equipped therein, and it is preferable that each of the compartments 111, 112, 113, 114, and 115 has a structure that allows the upper sides thereof to communicate one another so that the magnetic beads 102 may be transported from the first compartment 111 to the rearmost compartment (in the present exemplary embodiment, a fifth compartment 115 is illustrated and described as the rearmost compartment) by a designated magnetic member 103.

[0027] The reagent supply unit 300 has reagent chambers 311, 312, 313, 314, and 315 formed in a row, each having a perforated lower part thereof, corresponding to the respective compartments in order to supply a reagent (and a wash solution) to each compartment 111, 112, 113, 114, and 115 within the main body 100, and each of the reagent chambers 311, 312, 313, 314, and 315 has a built-in perforation pin (not illustrated) provided therein for bursting a capsule containing the reagent (and the wash solution). In the present disclosure, the reagent supply unit 300 may be configured such that each of the reagent chambers 311, 312, 313, 314, and 315 has an incline of about 45 degrees in order for the reagent flowing out from the reagent capsule (not illustrated) to flow better into the interior of the main body 100. The reagent supply unit 300 has a sample injection port 301 formed corresponding to the sample supply path 101 at a front of the first reagent chamber 311 corresponding to the first compartment 111 of the main body 100 in order to inject the sample into the first compartment 111.

[0028] The reaction chamber 200 receives a supply of the nucleic acid-containing reagent from the main body 100 by using vacuum pressure of the syringe 400, so as to amplify the nucleic acid, and is coupled and fixed horizontally to the side of the main body 100. To fasten the reaction chamber 200, a stand 141 for supporting the lower end of the reaction chamber 200 is formed on the side of the main body 100, and hooking grooves 142 corresponding to hooking projections 220 formed on opposite 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 is formed with a flow path 210 therein passing through a plurality of chambers and a probe (see FIG. 4b), and the end of the syringe 400 is connected to an outlet 211 of this flow path 210 (hereinafter referred to as a "flow path outlet"), and a reagent discharge pipe 500 extending from the rearmost compartment 115 within the main body is connected to an inlet 212 of the flow path (hereinafter referred to as a "flow path inlet") (see FIG. 3b).

[0030] The syringe 400 may be detachably mounted on a holder 120 formed on the side of the main body 100, and may generate a fluid-pushing pressure so as to fill the reaction chamber 200 with the reagent.

[0031] The reagent discharge pipe 500 is for transporting the reagent containing the nucleic acids extracted from the sample to the reaction chamber 200, and has a ∩-shaped cross-sectional structure connecting the rearmost compartment 115 of the main body 100 and the flow path inlet 212 of the reaction chamber 200 to each other, and may be provided with slide grooves 510 on opposite sides thereof so as to be fitted and fixed in a side cutout part 130 of the main body 100.

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

[0033] In addition, ultrasonic waves may be provided to the reaction chamber 200 in order to unwind two-stranded DNA into single strands in the polymerase chain reaction process, and to this end, a second ultrasonic generator 700 (see FIG. 3b) may be provided in contact with an aluminum film 230 sealed at the lower end part of the reaction chamber 200.

[0034] Having the above-described configuration according to the present disclosure, the diagnostic cartridge may be used as illustrated in FIG. 5.

[0035] First, when the collected sample is injected through the sample injection port 301 of the reagent supply unit 300, the sample flows into the interior of the first compartment 111 through the sample supply path 101 of the cartridge body 100, and a reagent capsule (not illustrated) is burst from a first reagent chamber 311 of the reagent supply unit 300 to fill the interior of the first compartment 111 with a reagent. In addition, a plurality of magnetic beads 102 is inserted into the interior of the first compartment 111, and then the magnetic beads 102 are moved by using a magnetic member 103 from the outside, so as to pulverize the sample and destroy the cells, thereby ensuring that the reagent and the sample are well mixed. Through such a process, components released from the cells of the sample may adhere to the surface of each magnetic bead 102.

[0036] In the illustrated configuration of the present disclosure, a wash solution for washing may be supplied by bursting a capsule through each of the second reagent chamber 312, the third reagent chamber 313, and the fourth reagent chamber 314 of the reagent supply unit 300, and the wash solution flowing out from the second, third, and fourth reagent chambers 312, 313, and 314 is filled inside the second compartment 112, the third compartment 113, and the fourth compartment 114, respectively.

[0037] In this state, the magnetic beads 102 inside the first compartment 111 are moved to the second compartment 112, the third compartment 113, and the fourth compartment 114 in a sequential manner by using the magnetic member 103, whereby foreign substances on the surface of each magnetic bead 102 are removed through the washing.

[0038] In addition, an elution solution for the nucleic acid extraction may be supplied by bursting a capsule in the fifth reagent chamber 315 of the reagent supply unit 300, and the elution solution flowing out from the fifth reagent chamber 315 is filled inside the fifth compartment 115, which is the rearmost compartment. In this case, when the washed magnetic beads 102 inside the fourth compartment 114 are moved into the fifth compartment 115 by using the magnetic member 103, the nucleic acids are extracted from the surface of each magnetic bead 102 by the elution solution.

[0039] The extracted nucleic acids are introduced into the reaction chamber 200 through the ∩-shaped reagent discharge pipe 500 by the vacuum pressure generated according to the suction stroke of the syringe 400 outside the main body 100, and then as the extracted nucleic acids move along the flow path 210, the nucleic acid amplification is performed.

[0040] The diagnostic cartridge of the present disclosure, which operates in this manner, provides an advantage that the beads with the adhered nucleic acids are transported by using the magnetic member inside the cartridge body, so the sample does not leak out of the cartridge, thereby performing more safe diagnostic work.

[0041] The following describes yet another exemplary embodiments of a diagnostic cartridge for extraction, amplification, and analysis of nucleic acids of the present disclosure with reference to FIGS. 6 to 13.

[0042] As illustrated in FIG. 6, the diagnostic cartridge for the extraction, amplification, and analysis of the nucleic acids of the present disclosure is configured to include: a cartridge body 100' (hereinafter also referred to as a "main body") with an open upper part thereof for extracting the nucleic acids from a sample; a reagent supply unit 300' inserted through the open upper part of the main body 100' and seated on the inner upper part of the main body 100'; and a reaction chamber 200' coupled to a side of the main body 100' and performing the nucleic acid amplification.

[0043] FIGS. 7a and 7b illustrate the shape and structure of the cartridge body illustrated in FIG. 6. The main body 100' has the open upper part thereof for mounting the reagent supply unit 300', and a plurality of hooking grooves 104a and 104b for mounting the reagent supply unit 300' is formed at predetermined intervals vertically on the upper parts of the front and rear walls. In addition, a plurality of compartments 111, 112, 113, 114, 115,..., etc. for accommodating the supplied reagent and sample is formed in a row inside the main body 100' below the reagent supply unit 300' seated on the hooking grooves 104a and 104b.

[0044] In the present exemplary embodiment, it is illustrated as having five compartments 111, 112, 113, 114, and 115 as an example through FIGS. 7a to 7b. Hereinafter, the fifth compartment 115 will be referred to as the "rearmost compartment".

[0045] The upper parts of the plurality of compartments 111, 112, 113, 114, are 115 are closed by the mounted reagent supply unit 300', and the sample supply path 101 for introducing a sample is formed at a front of the first compartment 111 among the sequentially arranged compartments 111, 112, 113, 114, and 115, and is connected to the first compartment 111. The first compartment 111 has a plurality of magnetic beads 102 (see FIG. 11a) equipped therein, and it is preferable that each of the compartments 111, 112, 113, 114, and 115 has a structure that allows the upper parts thereof to be opened and communicate one another between the compartments 111, 112, 113, 114, and 115 so that the magnetic beads 102 may be transported from the first compartment 111 to the rearmost compartment by a designated magnetic member 103. For example, such a communication structure may be realized by providing a recess part (i.e., a communication concave part) 108 formed at least partially at a lower height than a partition wall 105 positioned between each of the compartments 111, 112, 113, 114, and 115.

[0046] FIGS. 8a and 8c are perspective views illustrating the shape and structure of a reagent supply unit provided in the diagnostic cartridge of FIG. 6. FIGS. 9a and 9b are partial cutaway perspective views illustrating the initial state of the diagnostic cartridge mounted with the reagent supply unit of FIG. 6. According to FIGS. 8b and 8c, it can be seen that the reagent supply unit 300' has a number of reagent chambers 311, 312, 313, 314, and 315, which are formed in a row therein, corresponding to the number of compartments 111, 112, 113, 114, and 115 in order to supply the reagent (or the wash solution) to each of the compartments 111, 112, 113, 114, and115) in the main body 100'.

[0047] Each of such reagent chambers 311, 312, 313, 314, and 315 is formed obliquely downward toward a rear side from the inside of the reagent supply unit 300', and has a structure in which a lower part thereof is open but an upper end thereof is closed by the upper surface of the reagent supply unit 300'. In a state of being filled with the reagent (or the wash solution) inside, each of the reagent chamber 311, 312, 313, 314, and 315 may be provided with an open lower part thereof sealed by a designated film material (i.e., a sealing film) 310a (see FIG. 9b).

[0048] Meanwhile, in order to supply the reagent (or the wash solution) into the interior of each compartment 111, 112, 113, 114, and 115 of the main body 100' by perforating such a sealing film 310a, the cartridge main body 100' has a perforation pin 107 (see FIG. 7b) that corresponds to the sealing film 310a of each reagent chamber 311, 312, 313, 314, and 315 and protrudes upward sharply in each compartment 111, 112, 113, 114, and 115.

[0049] Referring to FIGS. 8b and 9b, each of the reagent chambers 311, 312, 313, 314, and 315 filled with the reagent (or the wash solution) has a structure extending obliquely downward from the closed upper surface to the rear, and as described above, the sealing film 310a is attached to an opening at the lower end of each reagent chamber, so as to prevent the reagent (or the wash solution) from flowing out. According to the illustrated drawing, it is preferable that each reagent chamber 311, 312, 313, 314, and 315 has an incline of about 45 degrees toward the front of the main body 100' from the inside of the reagent supply unit 300', and as each of the reagent chambers 311, 312, 313, 314, and 315 have the incline of about 45 degrees, the lower end of each reagent chamber 311, 312, 313, 314, and 315 sealed by the sealing film 310a also has an incline of about 45 degrees from the horizontal (see FIG. 8c). As described above, as each of the reagent chambers 311, 312, 313, 314, and 315 has the 45-degree inclination angle, the reagent flows in a direction toward a point along the incline when the sealing film 310a is perforated, so as to prevent the residual reagent from remaining inside each reagent chamber (see FIG. 8c).

[0050] In addition, it may be preferable that at least part of a rear wall surface 106 of each compartment 111, 112, 113, 114, and 115 of the main body 100' has an inclination angle of about 45 degrees so that the reagent supplied from the reagent chambers 311, 312, 313, 314, and 315 at the inclination angle of about 45 degrees may flow down smoothly.

[0051] In the present exemplary embodiment, FIG. 8a is a front perspective view of the reagent supply unit, and FIG. 8b is a rear perspective view of the reagent supply unit, respectively. At one end of the reagent supply unit 300', preferably in the front direction of the first reagent chamber 311 among the reagent chambers 311, 312, 313, 314, and 315, a sample injection port 301 is formed to inject the sample into the first compartment 111 in the main body 100'.

[0052] The sample injection port 301 extends downward so as to be positioned near the sample supply path 101 of the main body 100' , and an opening / closing door 302 openable and closable for sample injection may be provided at an inlet of the sample injection port 301. The opening / closing door 302 may be configured to slide along guides 303 facing each other at a predetermined interval on a front upper surface of the reagent supply unit 300', and to this end, guide grooves 302a to be fitted into and mounted in the guides 303 are formed on opposite sides of the opening / closing door 302.

[0053] In addition, at the other end of the reagent supply unit 300', that is, preferably on the upper surface of the reagent supply unit 300' in the same line as the fifth reagent chamber 315 (in the present exemplary embodiment, the fifth reagent chamber 315 is illustrated and described as the "rearmost reagent chamber") among the reagent chambers 311, 312, 313, 314, and 315, a nucleic acid extraction port 305 for extracting the nucleic acids from the interior of the rearmost compartment 115 in the main body 100' passes through the interior of the reagent supply unit 300' and extends downward to correspond to the rearmost compartment 115.

[0054] An inlet of the nucleic acid extraction port 305 may be provided with an opening / closing door 306 openable and closable for the nucleic acid extraction, and such an opening / closing door 306 may be configured to slide along guides 307 facing each other at a predetermined interval on an upper surface of the rearmost reagent chamber 315. To this end, guide grooves 306a to be fitted into and mounted on the guides 307 are formed on opposite sides of the opening / closing door 306.

[0055] Each of the above-described opening / closing doors 302 and 306 may have a shape with a handle protruding from the upper surface thereof so as to facilitate opening and closing by a user. It is preferable that stop protrusions 303a and 307a for the respective opening / closing doors 302 and 306 are formed at the front ends of the guides 303 and 307 through which the opening / closing doors 302 and 306 slide, i.e., at the ends of the sample injection port 301 side and the nucleic acid extraction port 305 side, and that fitting grooves 302b and 306b to be respectively fitted into and fixed to the stop protrusions 303a and 307a (see FIGS. 8b and 8c) are formed at the front ends of the opening / closing doors 302 and 306.

[0056] Hooking projections 308 for allowing the reagent supply unit 300' to be seated in a hooked state in the hooking grooves 104a and 104b of the main body 100' protrude on the front and rear surfaces of the reagent supply unit 300'. The undescribed symbol 304 in the drawing indicates a space part formed by thin-walling in the reagent supply unit 300'.

[0057] The reaction chamber 200' receives a supply of the nucleic acid-containing reagent from the main body 100 through vacuum pressure generated by using a syringe 400 so as to amplify the nucleic acid. In the present exemplary embodiment, the description will focus on a combined structure as illustrated in FIG. 6, in which the reaction chamber 200' is horizontally fitted to the side of the main body 100'.

[0058] To mount the reaction chamber 200', a mounting groove 142 for fitting and fixing one end of the horizontal surface of the reaction chamber 200' is formed on the side of the main body 100', preferably on the side toward the rearmost compartment 115 of the main body 100'.

[0059] Hereinafter, the reaction chamber 200' is described as having a structure in which one end of the horizontal surface thereof is fitted and fixed into the mounting groove 142 formed on the side of the main body 100', but is not limited thereto, and as illustrated in FIG. 1b of the previous exemplary embodiment, the reaction chamber 200' may also be provided with a structure in which hooking projections 220 are formed on the opposite sides of the front end of the reaction chamber 200 and hooking grooves 142 are formed on the upper left and right sides of the stand 141 formed on the side of the main body 100 so that when the reaction chamber 200 is coupled, the hooking projections 220 are fitted into and coupled to the hooking grooves 142.

[0060] The internal structure of the reaction chamber 200' may be provided in the same form as described in the previous exemplary embodiment. That is, as illustrated in FIG. 4b, the structure may be provided such that a flow path 210 passing through a plurality of chambers and a probe is formed in the reaction chamber 200, the end of the syringe 400 is connected to an outlet 211 of the flow path 210 (hereinafter referred to as a "flow path outlet"), and a reagent discharge pipe 500 extending from the rearmost compartment 115 in the main body is connected to an inlet 212 of the flow path (hereinafter referred to as a "flow path inlet") (see FIG. 5).

[0061] In the present exemplary embodiment, the syringe 400 may be detachably mounted on the holder 120 formed on the side of the main body 100', and may generate a negative suction pressure, i.e., a fluid-pushing pressure so as to be able to fill the reaction chamber 200' with the reagent.

[0062] In addition, in the present exemplary embodiment, the diagnostic cartridge further includes a reagent discharge pipe 500 configured to connect the rearmost compartment 115 of the main body 100 and the reaction chamber 200' to each other. The reagent discharge pipe 500 is for transporting the reagent containing the nucleic acids extracted from the sample to the reaction chamber 200, and has an exhaust path 501 formed therein in a ∩-shaped cross-sectional structure connecting the rearmost compartment 115 of the main body 100 and the flow path inlet 212 of the reaction chamber 200 to each other. The reagent discharge pipe 500 has a structure in which slide grooves 510 are formed on opposite sides thereof so as to be fitted and fixed in a side cutout part 130 of the main body 100.

[0063] FIGS. 10a to 10c illustrate the change in state of the diagnostic cartridge when the reagent supply unit 300' mounted on the upper part of the main body in FIG. 9 is pressed down and lowered. Referring to FIGS. 9a and 9b, in the initial state, the reagent supply unit 300' is positioned on the inner upper part of the main body 100', and in this state, the hooking projections 308 protruding from the front and rear surfaces of the reagent supply unit 300' are seated in the hooked state in upper hooking grooves 104a formed on the front and rear surfaces of the main body 100', thereby supporting the reagent supply unit 300'. In addition, in this case, it can be seen that a perforation pin 107 formed in each compartment 111, 112, 113, 114, and 115 of the main body 100' is maintained at a predetermined distance downward from the sealing film 310a attached to the lower end surface of each of the reagent chambers 311, 312, 313, 314, and 315.

[0064] Thereafter, when the reagent supply unit 300' is pressed down to lower the reagent chambers 311, 312, 313, 314, and 315 as illustrated in FIGS. 10a and 10b, the hooking projections 308 protruding from the front and rear surfaces of the reagent supply unit 300' are also lowered down and are seated in the hooked state in lower hooking grooves 104b formed on the front and rear surfaces of the main body 100', thereby supporting the reagent supply unit 300'.

[0065] In addition, in this case, as the reagent chambers 311, 312, 313, 314, and 315 descend, the sealing film 310a blocking the lower end surface of each reagent chamber 311, 312, 313, 314, and 315 is perforated by the perforation pin 107 formed in each compartment 111, 112, 113, 114, and 115 of the main body 100' as illustrated in FIG. 10c, and accordingly, the reagent (or the wash solution) filled in the reagent chambers 311, 312, 313, 314, and 315 may flow along the rear wall 106 inside the main body 100' and be supplied to the interior of each compartment 111, 112, 113, 114, and 115.

[0066] The diagnostic cartridge of the present disclosure having the configuration as described above may be used as illustrated in FIGS. 11a and 11b.

[0067] FIGS. 11a and 11b are cross-sectional example diagrams illustrating the movement principle and path of the reagent and nucleic acids in the diagnostic cartridge of FIG. 6. First, the sample is collected from a human body, and when the collected sample is injected through the sample injection port 301 of the reagent supply unit 300', the sample is introduced into the first compartment 111 through the sample supply path 101 of the main body 100'.

[0068] In this case, when the reagent supply unit 300' mounted on the inner upper part of the main body 100' is pressed down and lowered, the sealing film 310a blocking the lower surface of each reagent chamber 311, 312, 313, 314, and 315 is perforated by the perforation pin 107 of each compartment 111, 112, 113, 114, and 115 in the main body 100' (see FIG. 10c), and accordingly, the reagent (or the wash solution) flows down from each reagent chamber 311, 312, 313, 314, and 315 to fill the interior of each compartment 111, 112, 113, 114, and 115.

[0069] The plurality of magnetic beads 102 is inserted into the first compartment 111, and then the magnetic beads 102 are moved by using the magnetic member (i.e., the magnet) 103 from the outside, so as to pulverize the sample and destroy the cells, thereby ensuring that the reagent and the sample are well mixed. Through such a process, components released from the cells of the sample may adhere to the surface of each magnetic bead 102.

[0070] In the present exemplary embodiment, the second, third, and fourth reagent chambers 312, 313, and 314 of the reagent supply unit 300' accommodate the wash solution for washing, and as the sealing films 310a are perforated by the respective perforation pins 107 by the reagent supply unit 300' being lowered described above, the wash solution in the second, third, and fourth reagent chambers 312, 313, and 314 fills the interior of the second, third, and fourth compartments 112, 113, and 114. In this state, as the magnetic beads 102 inside the first compartment 111 are moved to the second, third, and fourth compartments 112, 113, and 114 in a sequential manner by using the magnetic member 103, foreign substances on the surface of each magnetic bead 102 are removed through a washing process using the wash solution.

[0071] In addition, in the present exemplary embodiment, the rearmost reagent chamber 315 of the reagent supply unit 300' accommodates an elution solution for nucleic acid extraction. In the same manner, as the sealing film 310a is perforated by the perforation pin 107 by the reagent supply unit 300' being lowered described above, the elution solution in the rearmost reagent chamber 315 fills the interior of the fifth compartment 115, which is the rearmost compartment in the main body 100'. In this case, when the washed magnetic beads 102 inside the fourth compartment 114 are moved into the rearmost compartment 115 by using the magnetic member 103, a nucleic acid (i.e., DNA) is extracted from the surface of each magnetic bead 102 by the elution solution.

[0072] In this case, a method of transporting an extracted nucleic acid (DNA) from above the precipitated magnetic beads 102 to above the reagent water level is applied, and the extracted nucleic acid may be discharged into the reaction chamber 200' through the ∩-shaped discharge path 501 in the reagent discharge pipe 500 by the vacuum pressure generated according to the suction stroke of the syringe 400 attached to the side of the main body 100. As the nucleic acid (DNA) moves along the flow path 210 after being introduced into the reaction chamber 200', the nucleic acid amplification is performed.

[0073] In general, several factors affect the amplification efficiency of PCR. The common factors include time, temperature, the amount and nature of polymerase, a DNA amount, dNPT, Mg2+, etc. Among them, when the purified DNA is transported to the reaction chamber 200', it is required to separate DNA from the magnetic beads 102, which are holding the DNA from the extraction stage. To this end, in the present exemplary embodiment, the magnet 103 is used to catch the beads 102.

[0074] In addition, when the purified DNA is transported to the reaction chamber 200', the remaining amount of reagent not attached to the magnet may flow into the reaction chamber 200'. In the present exemplary embodiment, in order to prevent this issue from occurring, it is preferable that the inlet of the ∩-shaped discharge path 501 in the reagent discharge pipe 500 be positioned at a higher water level than the beads 102 so that the DNA is suctioned in at a higher water level than the precipitated beads 102.

[0075] The diagnostic cartridge having the configuration and operation described above according to the present disclosure has an advantage that the diagnostic cartridge can be used for immunological diagnosis as well as molecular diagnosis. In particular, the diagnostic cartridge has an advantage that the magnetic beads with the attached nucleic acids are transported by using the magnet inside the cartridge body, so the sample does not leak out of the cartridge, thereby performing more safe diagnostic work.

[0076] FIGS. 12a and 12b illustrate examples of use for extracting purified DNA within the diagnostic cartridge of the present disclosure. Here, a pipette (or a syringe) P may be used to extract the purified DNA within the fifth compartment 115, which is the rearmost compartment.

[0077] In general, since conventionally developed cartridge for Point of Care Testing (POCT) test equipment is used for one-time testing purposes, the purified DNA obtained through the process within the cartridge is unable to be taken out of the cartridge. Accordingly, in a case where the test fails in the POCT equipment, there is the inconvenience of having to obtain the sample again because the purified DNA in the cartridge cannot be used continuously.

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

[0079] In this way, even though the test fails on the POCT test equipment, retesting is available without having to obtain a sample repeatedly as the case in the past because the extracted DNA is the same patient's DNA. Naturally, the extracted DNA may be used for other purposes as well.

[0080] FIGS. 13a and 13b illustrate yet another exemplary embodiment of a diagnostic cartridge according to the present disclosure. According to the present disclosure, a reaction chamber 200" may be formed integrally with a cartridge body 100" without the need for the configuration such as the stand 141 and the hooking grooves 142, which are illustrated in the configurations of FIGS. 1 and 6, so that an internal flow path 210 and the rearmost compartment 115' of the cartridge body may be configured to communicate with each other. A syringe 400' may be provided in a structure in which the surface of a cylinder 410 is formed integrally with the cartridge body 100" while the end of the cylinder 410 is integrally connected to a flow path outlet 211 of the reaction chamber 200".

[0081] In particular, in the present exemplary embodiment, the diagnostic cartridge may be formed by excluding the reagent discharge pipe 500 from the configurations of FIGS. 1 and 6, and raising the bottom height of the rearmost compartment 115' of the cartridge body 100" to have the same height as the reaction chamber 200", and in this way, the flow path 210 of the rearmost compartment 115' and the reaction chamber 200" can communicate to each other without the reagent discharge pipe 500 (see FIGS. 1 and 6).

[0082] FIGS. 14a and 14b are enlarged views illustrating a modified internal structure of the cartridge body provided in the diagnostic cartridge of FIG. 13. A panel-shaped perforation plate 107' in the present exemplary embodiment may be configured in place of the perforation pin 107 having a shape that protrudes sharply upward corresponding to the sealing film 310a of each reagent chamber 311, 312, 313, 314, and 315 in the configuration of FIG. 7b. Preferably, such perforation plates 107' may be formed of at least one pair of panels arranged in parallel and spaced apart from each other by a predetermined distance.

[0083] FIGS. 15 and 16 are partial cutaway perspective views illustrating the change in state of the diagnostic cartridge before and after the lowering of the reagent supply unit. In the initial state of FIG. 15, the reagent supply unit 300' is positioned at the inner upper part of the main body 100', and in this state, the hooking projections 308' protruding from the left and right surfaces of the reagent supply unit 300' are seated in a hooked state on the hooking protrusions 104c formed on the left and right surfaces of the main body 100', thereby supporting the reagent supply unit 300'. In addition, in this case, it can be seen that the perforation plate 107' formed in each compartment 111, 112, 113, 114, and 115 of the main body 100' is maintained at a predetermined distance downward from the sealing film 310a attached to the lower end surface of each of the reagent chambers 311, 312, 313, 314, and 315.

[0084] Thereafter, when the reagent supply unit 300' is pressed down to lower the reagent chambers 311, 312, 313, 314, and 315 as illustrated in FIG. 16a, the sealing film 310a blocking the lower end surface of each reagent chamber 311, 312, 313, 314, and 315 is perforated by the perforation plate 107' formed in each compartment 111, 112, 113, 114, and115) of the main body 100', whereby the reagent (or the wash solution) filled in the reagent chambers 311, 312, 313, 314, and 315 may flow down along the rear wall 106 inside the main body 100' and be supplied to the interior of each compartment 111, 112, 113, 114, and 115.

[0085] FIG. 17 is a cross-sectional example diagram illustrating the movement principle and path of the reagent and nucleic acids in the diagnostic cartridge of FIG. 13. After the sample collected from a human body is injected into the first compartment 111 through the sample injection port 301 of the reagent supply unit 300', the reagent supply unit 300' is pressed down to fill the interior of each compartment 111, 112, 113, 114, and 115' with the reagent (or the wash solution) from each reagent chamber 311, 312, 313, 314, and 315, respectively as illustrated in FIG. 16, and then the plurality of magnetic beads 102 is injected into the interior of the first compartment 111, and the magnetic member (the magnet) 103 is used from the outside to move the magnetic beads 102, so as to pulverize the sample and destroy the cells, thereby ensuring that the reagent and the sample are well mixed. Through such a process, the components released from the cells of the sample may adhere to the surface of each bead 102.

[0086] In the present exemplary embodiment, the second, third, and fourth reagent chambers 312, 313, and 314 of the reagent supply unit 300' accommodate the wash solution for washing, and as the sealing films 310a are perforated by the respective perforation plates 107' by the reagent supply unit 300' being lowered described above, the wash solution in the second, third, and fourth reagent chambers 312, 313, and 314 fills the interior of each of the second, third, and fourth compartments 112, 113, and 114. In this state, as the magnetic beads 102 inside the first compartment 111 are moved to the second, third, and fourth compartments 112, 113, and 114 in a sequential manner by using the magnetic member 103, foreign substances on the surface of each magnetic bead 102 are removed through the washing process using the wash solution.

[0087] In addition, in the present exemplary embodiment, the rearmost reagent chamber 315 of the reagent supply unit 300' accommodates an elution solution for the nucleic acid extraction. In the same manner, as the sealing film 310a is perforated by the perforation plate 107' by the reagent supply unit 300' being lowered described above, the elution solution in the rearmost reagent chamber 315 fills the interior of the fifth compartment 115, which is the rearmost compartment in the main body 100'. In this case, when the washed magnetic beads 102 inside the fourth compartment 114 are moved into the rearmost compartment 115' by using the magnetic member 103, a nucleic acid (DNA) is extracted from the surface of each magnetic bead 102 by the elution solution.

[0088] In this case, the extracted nucleic acid (DNA) may be discharged into the reaction chamber 200' by the vacuum pressure generated according to the suction stroke of the syringe 400 integrally coupled to the flow path outlet 211 of the reaction chamber, and as the nucleic acid (DNA) moves along the flow path 210 after being introduced into the reaction chamber 200', the nucleic acid amplification is performed.

[0089] FIG. 18 illustrates the configuration and use state of POCT test equipment configured to emit ultrasonic waves for a reaction chamber mounted on a diagnostic cartridge according to one exemplary embodiment of the present disclosure. The ultrasonic waves may be provided to the reaction chamber 200' in order to unwind two-stranded DNA into single strands in a polymerase chain reaction process. To this end, a reaction unit in the reaction chamber 200' accommodating a DNA-including reagent is sealed with a metal film, e.g., an aluminum film 230, and an ultrasonic generator 700' is placed in contact with the bottom of the aluminum film 230.

[0090] Such an ultrasonic generator 700' raises the temperature to 95°C to denature and inactivate most of DNA. When the temperature of a section for the denaturation and inactivation during the DNA amplification process can be lowered or eliminated, the time taken in the existing PCR amplification process in which temperature cycles are repeated may be significantly reduced.

[0091] Although various exemplary embodiments of the present disclosure are described above, the contents described so far are only to the extent of exemplifying some of the preferred exemplary embodiments of the present disclosure, and are not limited by the above-described contents except as may appear in the claims attached below. Therefore, it should be understood that many changes, modifications, and substitutions of equivalents may be made by those skilled in the art without departing from the technical idea and gist of the exemplary embodiments of the present disclosure within the scope set forth in the claims below.

Claims

1. A diagnostic cartridge for extraction, amplification, and analysis of nucleic acids, the diagnostic cartridge comprising: a cartridge body having a plurality of compartments formed in a row therein in order to accommodate a supplied reagent and a supplied sample, having each of the plurality of compartments open at an upper part side thereof so as to enable reagent supply, and having a sample supply path formed at a front of a first compartment among the plurality of compartments in order to introduce the sample into the first compartment; a reagent supply unit configured to contain at least the reagent (or a wash solution) and to be inserted and seated through the open upper part of the cartridge body; and a reaction chamber provided on one side of the cartridge body to perform the nucleic acid amplification.

2. The diagnostic cartridge of claim 1, wherein each partition wall between the plurality of compartments of the cartridge body has a part thereof formed at a height lower than surrounding components, so as to allow a magnetic member operating outside the cartridge body to transport the nucleic acids adhered to magnetic beads within one compartment to another adjacent compartment.

3. The diagnostic cartridge of claim 1, wherein the reaction chamber is formed integrally with the cartridge body.

4. The diagnostic cartridge of claim 1, wherein one or more perforation pins or perforation plates are built in an interior of each compartment of the cartridge body in order to perforate a sealing film at a lower end of the reagent supply unit when the reagent supply unit is lowered.

5. The diagnostic cartridge of claim 1, further comprising: a designated suction means for generating vacuum pressure according to a predetermined suction stroke so as to enable the nucleic acids to move along a flow path within the reaction chamber from the cartridge body, wherein a bottom surface of a rearmost compartment and an inlet of the flow path in the reaction chamber are formed at the same height so as to allow the rearmost compartment of the cartridge body to communicate with the flow path in the reaction chamber in order to discharge a nucleic acid extracted from the cartridge body into the reaction chamber.

6. The diagnostic cartridge of claim 1, wherein the reagent supply unit has a plurality of reagent chambers formed in a row and each provided with a lower end thereof sealed with a sealing film corresponding to each compartment of the cartridge body in a state of accommodating the reagent (and the wash solution) within the reagent supply unit in order to supply at least the reagent (or the wash solution) to each compartment within the cartridge body, and the lower end of each of the plurality of reagent chambers has an incline of at least 45 degrees.

7. The diagnostic cartridge of claim 6, wherein in the reagent supply unit, a sample injection port for injecting the sample into the first compartment is formed, corresponding to the sample supply path, at a front of a first reagent chamber corresponding to the first compartment among the plurality of compartments of the cartridge body.

8. The diagnostic cartridge of claim 6, wherein in the reagent supply unit, a nucleic acid extraction unit for extracting a purified nucleic acid from a rearmost compartment within the main body is formed on a rear end side corresponding to the rearmost compartment among the plurality of compartments of the main body, and the nucleic acid extraction through the nucleic acid extraction unit uses a pipette or a syringe.

9. The diagnostic cartridge of claim 1, further comprising: at least one ultrasonic generator provided in contact with the aluminum film attached to a lower end part of the main body, so as to provide ultrasonic waves for smooth mixing of the nucleic acids of the sample and the reagent in the first compartment among the plurality of compartments of the cartridge body.

10. The diagnostic cartridge of claim 1, wherein the reaction chamber further comprises: at least one ultrasonic generator provided in contact with an aluminum film sealing at least a part of the reaction chamber, so as to provide ultrasonic waves in order to unwind double-stranded DNA into single strands in a polymerase chain reaction process.

11. A diagnostic cartridge for extraction, amplification, and analysis of nucleic acids, the diagnostic cartridge comprising: a cartridge body with an open upper part thereof and for extracting the nucleic acids from a sample; a reagent supply unit inserted and seated through the open part of the cartridge body; a reaction chamber integrally formed on a side of the cartridge body and for performing the nucleic acid amplification; and a suction means connected to move an extracted nucleic acid from the cartridge body to the reaction chamber and connected to an outlet of a flow path within the reaction chamber, so as to fill the reaction chamber with the nucleic acid moved from the cartridge body by using vacuum pressure generated according to a suction stroke.

12. The diagnostic cartridge of claim 11, wherein in the plurality of compartments, a part of a partition wall between the plurality of compartments is configured to have a height lower than a reference height in a predetermined amount.

13. The diagnostic cartridge of claim 11, wherein the suction means is composed of a cylinder integrally formed with the cartridge body and a separate piston.

14. The diagnostic cartridge of claim 11, wherein the cartridge body further comprises: a pipette or a syringe as an extrusion means.

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