Patent Description:
In general, samples obtained from a human body or the body of animal are refined and then undergo a predetermined examination in a laboratory in some cases. In this case, generally, preprocessing and processing such as refinement are performed on a sample, and the refined sample may be finally collected as analyte and a predetermined test may be performed. As an example of an analyte collecting device and method, and an analyte inspection system, a device and method of refining nucleic acid and a system for examining the refined nucleic acid may be exemplified.

Refinement of nucleic acid, which is a necessary technique that is widely used in the fields of genetic engineering and molecular biology, is a very important technique in terms of study, medical treatment, and industry as a preprocessing stage for techniques such as Southern blot, Northern blot, and polymerase chain reaction (PCR). Such refinement of nucleic acid is conventionally achieved through chemical and physical method that use ultrasonic waves, heat, proteinase, alcohols, a special reagent, etc. and researchers perform a nucleic acid refinement process using a pipette. However, recently, methods of more conveniently refining nucleic acid using magnetic particles have been introduced in a wide filed. However, these methods should be performed in laboratories, require a large amount of time and manpower, and have limitation to be generally used.

A process of refining nucleic acid includes stages such as dissolving (lysis) of biomaterials such as a cell, nucleic acid-magnetic particle binding, washing, and elution, and requires reagents and treatments suitable for each of the stages. An analyte that has undergone such a refinement process can be collected by a predetermined amount and predetermined necessary tests can be performed on the analyte. That is, refined nucleic acid is moved into a transparent amplification and detection tube and is amplified by real-time PCR or similar techniques, whether there is pathogenic nucleic acid is optically detected using fluorescent labeling, and accordingly, it is possible to diagnose corresponding diseases.

An analyte collection device for refining and collecting a sample as an analyte in a predetermined quantity should require minimum manpower for the refinement process, should be filled with a predetermined solution for refinement, and should be small to secure mobility, in order to reduce power of a hospital and perform point-of-care testing (POCT). Further, disposability should be secured to prevent contamination by biomaterials, so the device needs to be implemented at a low cost. However, there are little study about an analyte collection device, a method using the analyte collection device, and a system for examining an analyte that completely satisfies those conditions.

<CIT> discloses a syringe device having a hollow tubular body that receives a plunger, having in combination a handle portion rotatable within a sealing member portion, for obtaining gas-free blood samples. The plunger includes a plunger handle and a resilient sealing member which are rotatably connected. Both the plunger handle and sealing member are received through an open trailing end of the tubular body. The sealing member is further adapted to slide along the interior surface of the main tubular body in a sealed relationship therewith. In its undeformed shape, the sealing member is generally cylindrical with a circular lip at either end and an intermediate lip disposed between the end lips. A pair of toroidal void spaces are defined between the cylindrical wall of the sealing member, the lips and the internal surface of the main tubular body.

<CIT> relates to a nucleic acid purification device, in which a washing container and an elution container are bonded to each other to form a channel for moving a nucleic acid. The washing container includes an outer peripheral wall which accommodates a connection portion of the first channel and a second channel, the elution container includes a plurality of flanges in the periphery of the second channel in contact with an inner wall of the outer peripheral wall, the plurality of flanges are arranged in a portion which is to be inserted into the inside of the outer peripheral wall of the elution container, and one space which is partitioned by two flanges adjacent to each other among the plurality of flanges and the outer peripheral wall communicates with another space adjacent to the one space in a state of being divided by one of the two flanges adjacent to each other.

Embodiments of the present disclosure provide an analyte collecting device that can be achieved at a low cost, can be achieved in a small size, and can efficiently process a sample through an automated process in order to refine and pre-process a sample using magnetic particles, and an analyte collecting method and a system for examining an analyte using the analyte collecting device.

In accordance with a first aspect of the present disclosure, there is provided an analyte collecting device including: a case including an opening and an internal space; and a piston including one or more partition walls dividing the internal space into a plurality of internal spaces, the piston being inserted into the internal space through the opening of the case to reciprocate in the internal space.

The case may include: an exhaust port formed at an end portion opposite to a side in which the piston is inserted such that the internal space communicates with an outside, a sample put in the internal space being discharged through the exhaust port to the outside of the case; and a blowback portion provided at the opposite end portion to the side in which the piston is inserted, the blowback portion including a flow hole formed such that both ends thereof communicate with the internal space.

The internal space may be filled with a solution containing a magnetic substance, and the sample input in the internal space may be discharged through the exhaust port as an analyte after undergoing predetermined processing by the solution in the internal space.

The one or more partition walls may include four partition walls, the plurality of internal spaces may include a first section, a second section, a third section, and a fourth section sequentially divided by the four partition walls, the first section to the fourth section being sequentially disposed away from the opening of the case, the first section may be filled with a solution that dissolves a biomaterial contained in the sample and binds at least a portion of an analyte in the biomaterial to the magnetic substance, the second section may be filled with a solution that washes at least a portion of the analyte bonded to the magnetic substance, the third section may be filled with a solution that elutes at least a portion of the analyte bonded to the magnetic substance from the magnetic substance, and the fourth section may be formed adjacent to the third section and in contact with an inner end of the case. the solution filled in the first section may include at least one of a lysis/binding buffer and isopropyl alcohol (<NUM>-propanol), the solution filled in the second section may include a washing buffer, and the solution filled in the third section may include an elution buffer.

The piston may include a center column. The one or more partition walls may include a plurality of partition walls spaced apart from each other and radially extend from the circumferential surface of the center column, and the internal space may be divided into a plurality of sections by the partition walls and at least some of the divided plurality of sections may be filled with different solutions.

The piston may further include: a flange attached to at least one of two surfaces of each of the one or more partition walls provided perpendicular to an insertion direction of the piston, the flange having a circumferential surface provided closer to an inner wall surrounding the internal surface of the case than the circumferential surface of each of the one or more partition walls; and a sealing member provided to surround the circumferential surface of each of the one or more partition walls and being in contact with the inner wall of the case.

The case may further include: a specimen injection port including an injection hole formed at the case to allow the internal space to communicate the outside of the case so that a sample can be injected. The specimen injection port may further include a cap sealing the injection hole by selectively covering the injection hole.

An aggregating groove may be recessed from an inner wall forming the internal space of the case, the internal space may be filled with a solution containing a magnetic substance, and the magnetic substance may be aggregated in the aggregating groove when a magnetic force is applied toward the aggregating groove from the outside.

The analyte that is collected by the analyte collecting device may include at least one of nucleic acid, protein, vesicle, lipid, a carbohydrate, a cell, and a substance separated therefrom.

In accordance with a second aspect of the present disclosure, there is provided a system for examining an analyte, the system including: an analyte collecting device including a case including an opening and an internal space, and a piston including one or more partition walls dividing the internal space into a plurality of internal spaces, the piston being inserted into the internal space through the opening of the case to reciprocate in the internal space; and a holder separably holding the analyte collecting device.

The system may further include: a plunger for translating the piston in the internal space by pushing or pulling a head of the piston; and a controller. The plunger may be controlled by the controller.

The internal space may be filled with a solution containing a magnetic substance, predetermined processing may be performed on a sample put in the case by the solution filled in the internal space, and the predetermined processing may include a plurality of stages which are sequentially performed as the controller controls operation of the plunger.

The case may include: an exhaust port formed at an end portion opposite to a side in which the piston is inserted such that the internal space communicates with an outside, a sample, which is put in the internal space and undergoes the predetermined processing, being discharged from the case as an analyte; and a blowback portion provided at the opposite end portion to the side in which the piston is inserted, the blowback portion including a flow hole formed such that both ends thereof communicate with the internal space. The controller may control the plunger to push the piston toward the blowback portion such that the analyte is discharged through the exhaust port by a blowback phenomenon.

The case may have an aggregating groove recessed from an inner wall forming the internal space, and the system may further include: an aggregating device configured to selectively apply a magnetic force toward the aggregating groove so that the magnetic substance is aggregated in the aggregating groove, the aggregating device being controlled by the controller.

The system may further include: a de-aggregating device configured to selectively apply a magnetic force to the internal space and being controlled by the controller. The controller may control the de-aggregating device with the magnetic substance aggregated in the aggregating groove so that a magnetic force is applied to the internal space and the magnetic substance aggregated in the aggregating groove is separated.

In accordance with a third aspect of the present disclosuer, there is provided a method of collecting analyte using an analyte collecting device including: a case including an opening and an internal space; and a piston including one or more partition walls dividing the internal space into a plurality of internal spaces, the piston being inserted into the internal space of the case to reciprocate in the internal space, the method including: putting a sample in the internal space; and collecting the sample as an analyte by sequentially performing a predetermined processing including a plurality of stages while the piston is moved in the internal space.

The case may include: an exhaust port formed at an end portion opposite to a side in which the piston is inserted such that the internal space communicates with an outside, the sample accommodated in the internal space being discharged through the exhaust port to the outside of the case; and a blowback portion provided at the opposite end portion to the side in which the piston is inserted, the blowback portion including a flow hole formed such that both ends thereof communicate with the internal space. The method may further include discharging the analyte in which the piston moving in the internal space pushes a section contained an analyte undergoing the predetermined processing toward the blowback portion formed at an end portion of the case, and the analyte pressed through the blowback portion is discharged through an exhaust port formed at the case.

An aggregating groove may be recessed from an inner wall forming the internal space of the case, and a magnetic substance is contained in a solution filled in the internal space. The method may further include aggregating the magnetic substance in the aggregating groove by applying a magnetic force to the aggregating groove.

The one or more partition walls may include four partition walls, the plurality of internal spaces include a first section, a second section, a third section, and a fourth section sequentially divided by the four partition walls, the first section of the four sections may be formed closest to an opening of the case, the second section may be formed adjacent to the first section with one of the partition walls therebetween, the third section may be formed adjacent to the second section with one of the partition walls therebetween, and the fourth section may be formed adjacent to the third section with one of the partition walls therebetween to be in contact with an inner end of the case, the sample may be put in the first section, and the sequentially performing the predetermined processing may further include: bonding at least a portion of an analyte in a biomaterial contained in the sample to the magnetic substance by dissolving the biomaterial by the solution filled in the first section, disposing the second section above the aggregating groove by moving the piston backward after the magnetic substance bound to at least a portion of the analyte is aggregated in the aggregating groove by applying a magnetic force to the aggregating groove, floating the magnetic substance in the second section in which the magnetic force applied to the aggregating groove is removed, and the magnetic substance aggregated in the aggregating groove is separated and floats into the second section by applying a magnetic force to the internal space; washing at least a portion of the analyte bound to the magnetic substance by a solution filled in the second section; disposing the third section above the aggregating groove by moving the piston backward after the magnetic substance bound to at least a portion of the analyte is aggregated in the aggregating groove by applying a magnetic force to the aggregating groove; floating the magnetic substance in the third section in which the magnetic force applied to the aggregating groove is removed, and the magnetic substance aggregated in the aggregating groove is separated and floats into the third section by applying a magnetic force to the internal space; eluting at least a portion of the analyte bound to the magnetic substance from the magnetic substance by a solution filled in the third section; and aggregating the magnetic substance with at least a portion of the analyte eluted in the aggregating groove by applying a magnetic force to the aggregating groove.

The solution filled in the first section may include at least one of a lysis/binding buffer and isopropyl alcohol (<NUM>-propanol), the solution filled in the second section includes a washing buffer, and the solution filled in the third section includes an elution buffer.

According to embodiments of the present disclosure, since processing such as refinement, amplification, detection can be automatically performed without intervention of a user after a sample is put in the device, there is an effect that usability is high, secondary infection of a user or a third party can be prevented when the sample is processed.

Further, since the analyte that finishes being refined can be automatically conveyed, a user does not intervene in movement of a solution for the next reaction, so there is an effect that sample-to-answer can be implemented and carryover contamination can be prevented.

Further, there is an effect that a yield ratio an analyte is prevented from changing depending on the skill of users and it is possible to achieve a constant yield ratio of an analyte every time.

Further, there is an effect that the structure of the device is simplified, the components can be minimized, the cost is reduced by downsizing, and spatial usability can be increased.

Further, since it is possible to add a plurality of refining stages in one device, there is an effect that multiple specimens can be processed and detected.

Hereinafter, specific embodiments will be described in detail with reference to the drawings.

In describing the present disclosure, detailed descriptions of known configurations or functions may be omitted to clarify the present disclosure.

When an element is referred to as being 'connected' to, or 'contacted' with another element, it should be understood that the element may be directly connected to, or contacted with another element, but that other elements may exist in the middle.

The terms used in the present disclosure are only used for describing specific embodiments, and are not intended to limit the present disclosure. Singular expressions include plural expressions unless the context clearly indicates otherwise.

Hereafter, an analyte collecting device and a system for examining an analyte according to an embodiment of the present disclosure are described with reference to the drawings.

Referring <FIG> and <FIG>, an analyte collecting device <NUM> according to an embodiment of the present disclosure may include, in a broad meaning, a case <NUM> and a piston <NUM>. The case <NUM> and the piston <NUM>, for example, may be made of any one material of plastic, rubber, ceramic, an inorganic compound, and metal, or a combination thereof. The case <NUM> and the piston <NUM>, for example, may be manufactured through processes such as blow molding, compression molding, extrusion molding, injection molding, laminating, reaction injection molding, matrix molding, rotational molding, spin casting, transfer molding, thermoforming, and 3D printing. Further, the case <NUM> and the piston <NUM> can be manufactured in a large quantity, for example, as disposables by an existing automated facility. Further, the case <NUM> and the piston <NUM> may be individually manufactured and assembled to be provided.

The case has a space <NUM> therein in which a sample can be put, a piston <NUM> may be inserted and assembled to each other in the internal space <NUM>. The internal space <NUM> of the case <NUM> may be provided in a shape with one side thereof opened and may be formed in a shape corresponding to the piston <NUM> so that the piston <NUM> can be inserted and reciprocated in the internal space <NUM>. Further, the internal space <NUM> of the case <NUM> may be divided into a plurality of sections by partition walls <NUM> of the piston <NUM>. For example, the internal space <NUM> of the case <NUM> may be divided into a total of four sections 102a, 102b, 102c, and 102d, but the present disclosure is not limited thereto.

A sample that is put into the internal space <NUM> may be liquid, solid, of a mixture including some or all of a cell, a virus, a tissue, exosome, protein, nucleic acid, an antigen, and an antibody, and for example, may be a specimen taken from a human body. When the sample that is put into the internal space <NUM> is a specimen taken from a human body, for example, the nucleic acid in cells existing in the sample may be refined using the analyte collecting device <NUM>.

Further, the internal space <NUM> may be filled with a solution containing a magnetic substance and the plurality of sections may be filled with different solutions. For example, when the internal space <NUM> is divided into a total of four sections 102a, 102b, 102c, and 102d, the first section 102a may be formed closest to the case <NUM> formed such that the piston <NUM> can be inserted therein, among the four sections 102a, 102b, 102c, and 102d, and the first section 102a may be filled with a solution for binding at least a portion of the analyte, which is in a biomaterial contained in the sample, to a magnetic substance through dissolution of the biomaterial.

For example, an analyte that is collected by the analyte collecting device <NUM> may be nucleic acid, protein, vesicle (exosome, etc.), lipid, a carbohydrate, a cell (a blood cell, an immunocyte, an oncocyte, pathogenic bacteria, etc.) and may include a biomaterial itself contained in a sample or a substance that can be physically and/or chemically separated from the biomaterial. Further, for example, when the nucleic acid in cells existing in a sample is refined by the analyte collecting device <NUM>, the analyte that is collected by the analyte collecting device <NUM> may include nucleic acid.

The solution that fills the first section 102a may include, for example, at least one of a lysis/binding buffer and <NUM>-Propanol, and more particularly, may be provided as a solution including some or all of magnetic nano/micro particles, salts (ex. Tris-HCl), chelating agent (ex. ethylenediaminetetraacetic acid (EDTA)), a detergent (ex. Sodium dodecyl sulfate (SDS) and Triton X-<NUM>), a reductant (ex. Dithiothreitol (DTT), a chaotropic agent (ex. Guanidine thiocyanate), enzyme (ex. Proteinase K), alcohol (ex. <NUM>-Propanol), and distilled water.

Further, the second section 102b is formed adjacent to the first section 102a with one of the plurality of partition walls <NUM> therebetween, and the second section 102b may be filled with a solution that enables washing of at least a portion of the analyte bound to a magnetic substance.

The solution that fills the second section 102b, for example, may include washing buffer, and more particularly, may be provided as a solution including some or all of diethyl pyrocarbonate (DEPC), sodium citrate tribasic dehydrate, alcohol (ex. Ethanol, <NUM>-propanol), and distilled water.

Further, the third section 102c is formed adjacent to the second section 102b with one of the plurality of partition walls <NUM> therebetween, and the third section 102c may be filled with a solution that enables at least a portion of the analyte bound to a magnetic substance to be eluted from the magnetic substance.

The solution that fills the third section 102c, for example, may include an elution buffer, and more particularly, may be provided as a solution including some or all of salts (ex. Tris-HCl), a chelating agent (ex. Ethylenediaminetetraacetic acid (EDTA), diethyl pyrocarbonate (DEPC), and distilled water.

Further, the fourth section 102d may be formed adjacent to the third section 102c with one of the plurality of partition walls <NUM> therebetween and may be formed in contact with an inner end positioned opposite to the opening of the case <NUM>. The fourth section 102d may be filled with gas such as air.

Meanwhile, the case <NUM> may include a specimen injection port <NUM>, an aggregating groove <NUM>, a blowback portion <NUM>, an exhaust port <NUM>, a bed <NUM>, and wings <NUM>.

The specimen injection port <NUM> may include an injection hole <NUM> formed at the case <NUM> to connect the internal space <NUM> and the outside of the case <NUM> so that a sample can be injected, and may further include a cap <NUM> sealing the injection hole <NUM> by selectively covering the injection hole <NUM>.

The injection hole <NUM> may be formed in a shape of which the top is wide and is narrowed downward, and for example, may be formed on a conical shape, but the present disclosure should not be construed as being limited to the shape of the injection hole <NUM>. Further, the injection hole <NUM> may be recessed with respect to a top surface of the case <NUM> so that the lower end of the injection hole <NUM> may communicates with the internal space <NUM>. However, this is only an example and the injection hole <NUM> may be formed on a side surface or the bottom surface other than the top surface of the case <NUM>.

Further, as the piston <NUM> is moved in one direction in the internal space <NUM>, the lower end of the injection hole <NUM> can be sequentially communicated with the sections 102a, 102b, 102c, and 102d. When a sample is injected through the injection hole <NUM>, basically, the sample may be injected while the injection hole <NUM> communicates with the first section 102a, and the first stage of the refinement process may be performed immediately after the sample is injected into the first section 102a.

The cap <NUM> may be made of an elastic material, for example, rubber, and the lower end thereof is formed to correspond to the shape of the injection hole <NUM> such that when the lower end is inserted in the injection hole <NUM>, the cap can completely cover and seal the injection hole <NUM>. The cap <NUM> can prevent external foreign substances from permeating into the internal space <NUM> by sealing the injection hole <NUM> when the device is not used, and the cap <NUM> may be separated from the injection hole <NUM> and the injection hole <NUM> may be opened so that a sample can be injected into the injection hole <NUM>. After a sample is injected through the injection hole <NUM>, the cap <NUM> is coupled again, so the injection hole <NUM> can be sealed. Accordingly, it is possible to prevent foreign substances from permeating into the internal space not only before processing, but while processing is performed.

The aggregating groove <NUM> is recessed from an inner wall forming the internal space <NUM>, and when a magnetic force is applied toward the aggregating groove <NUM> from the outside, the magnetic substance in the internal space <NUM> can be aggregated in the aggregating groove <NUM>. In this case, dissolution and a binding action of the sample occur in the first section 102a, so when the magnetic substance and another biomaterial are bound, the magnetic substance and the material bound to the magnetic substance can be aggregated in the aggregating groove <NUM>.

Further, the aggregating groove <NUM> may be formed on an inner wall of the case <NUM> not to interfere with the injection hole <NUM>, and in this embodiment, the aggregating groove <NUM> is exemplified as being form on the bottom surface of the internal space <NUM> of the case, but the present disclosure is not limited thereto. For example, the aggregating groove <NUM> may be formed on a side surface or the top surface of the internal space <NUM>. Further, the aggregating groove <NUM> may be formed as a semispherical shape, but the present disclosure is not construed to be limitative due to the shape of the aggregating groove <NUM>, and the aggregating groove <NUM> may be formed in other shapes such as a conical shape and a hexahedron shape, depending on cases.

Further, the aggregating groove <NUM> may be formed on the same line as the injection hole <NUM>. Alternatively, the aggregating groove <NUM> may be formed at a position within a range in which the aggregating groove <NUM> and the injection hole <NUM> both can be communicated with at least the first section 102a. Accordingly, a sample injected through the injection hole <NUM> can be accommodated and aggregated in the aggregating groove <NUM> without additional movement of the piston <NUM>. However, this is only an example, and even if the aggregating groove <NUM> is formed at a position where the aggregating groove <NUM> cannot be communicated with both of the injection hole <NUM> and any one of the sections 102a, 102b, 102c, and 102d, a sample can be aggregated by the additional movement of the piston <NUM>.

The blowback portion <NUM> is provided at an end opposite to a side in which the piston <NUM> is inserted and includes a flow hole <NUM> having both ends communicated with the internal space. Further, the blowback portion <NUM> may be formed on the top of the case <NUM>, but the present disclosure is not limited thereto, and the blowback portion <NUM> may be formed on a side or the bottom of the case <NUM>. When the piston <NUM> is moved forward toward the blowback portion <NUM>, the gas such as air existing in the fourth section 102d is blown back to the flow hole <NUM> by the blowback portion <NUM>, so the analyte that finishes being refined and exists in the third section 102c can be discharged through the exhaust port <NUM> and collected in a collection container X (see, <FIG>).

To this end, the flow hole <NUM> includes a flow hole inlet <NUM>, a bridge <NUM>, and a flow hole outlet <NUM>. Each of the flow hole inlet <NUM> and the flow hole outlet <NUM> is formed such that an end thereof is communicated with the internal space <NUM>, and other end thereof is connected through the bridge <NUM>, so that the entire flow hole <NUM> may be formed as an U-shaped channel. In this case, the flow hole inlet <NUM> may be formed closer to the end opposite to the opening of the internal space <NUM> than the flow hole outlet <NUM>. Accordingly, when the piston <NUM> is moved forward to narrow the fourth section 102d, the gas such as air in the fourth section 102d flows into the flow hole inlet <NUM> by pressure, and can pass through the bridge <NUM> and the flow hole outlet <NUM> and then flow into the third section 102c adjacent to the fourth section 102d. The analyte accommodated in the third section 102c can be pushed through the exhaust port <NUM> by the pressure of the inflow gas and can be discharged from the case <NUM>. The analyte discharged as described above can be collected in the collection container X coupled to a container coupling protrusion <NUM> to be described below.

Due to the configuration of the blowback portion <NUM>, a user can finely adjust the amount of gas that is blown back through the flow hole <NUM> by adjusting the degree of pressing the piston <NUM>. As the amount of the gas that is blown back is adjusted, it is possible to finely adjust the amount of the analyte that is discharged through the exhaust port <NUM> and then collected. As described above, according to the present disclosure, since it is possible to finely adjust the amount of the collected analyte by finely adjusting the degree of pressing the piston <NUM>, the analyte collecting device <NUM> according to the present disclosure can be especially and usefully used when performing an examination in which it is very important to collect an analyte in a predetermined quantity.

Meanwhile, the flow hole <NUM> of the blowback portion <NUM> may be formed in a shape that is open such that the top of the bridge <NUM> is communicated with the outside. Accordingly, a cover <NUM> for selectively sealing the open surface of the bridge <NUM> may be further provided. The cover <NUM> may be made of an elastic material, for example, rubber, and the lower end thereof is formed to correspond to the shape of the bridge <NUM> such that when the lower end of the cover <NUM> is inserted in the bridge <NUM>, the cover can completely cover and seal the opening of the bridge1324. The cover <NUM> can prevent external foreign substances from permeating into the internal space <NUM> by sealing the flow hole <NUM>, whereby external foreign substances can be prevented from permeating into the internal space <NUM> not only before processing, but while processing is performed.

In this embodiment, it is exemplified that the flow hole <NUM> of the blowback portion <NUM> is formed in an open shape to be communicated with the outside and the opening of the flow hole <NUM> is sealed by the cover <NUM>, but the present disclosure is not limited thereto For example, the flow hole <NUM> may be formed in a shape such that the bridge <NUM> is not open and is only communicated with the internal space <NUM> without being communicated with the outside as it is.

The exhaust port <NUM> may be formed to be communicated with the outside at an end opposite to the opening in which the piston <NUM> is inserted, and may be formed such that a sample that has undergone predetermined processing in the internal space <NUM> can be discharged as an analyte from the case <NUM>. To this end, the exhaust port <NUM> may be formed through one side of the case <NUM>, and as shown in <FIG>, may be formed at a position facing the blowback portion <NUM>. However, this is only an example and the exhaust port <NUM> may be formed at a position not facing the blowback portion <NUM>. Further, in this embodiment, although it is exemplified that the exhaust port <NUM> is formed on the bottom of the internal space <NUM>, the present disclosure is not limited thereto and the exhaust port may be formed on a side or the top of the internal space <NUM>. In this case, when an analyte is discharged through the exhaust port <NUM>, a force by gravity is not applied but the analyte can be collected by the pressure due to the blowback phenomenon.

Further, the container coupling protrusion <NUM> may be formed at the portion where the exhaust port <NUM> of the case <NUM> is formed. The container coupling protrusion <NUM> may protrude from the outer surface of the case <NUM> and may be formed in a shape extending the exhaust port <NUM> to the outside of the case <NUM>. Further, the container coupling protrusion <NUM> may be formed in a shape that can be fastened to the collection container X, and the container coupling protrusion <NUM> and the collection container X can be fastened to each other by fitting, bolting, or the like. The collection container X may be made of soft plastic, but the present disclosure is not limited thereto.

The bed <NUM> may be formed in the bottom of the case <NUM> and may be held by a holder <NUM> to be described below so that the analyte collecting device <NUM> can be mounted thereon. Further, wings <NUM> may be formed a both sides of the bed <NUM>. The wings <NUM> may be formed in a shape protruding throughout the entire width of sides of the case <NUM> and support both sides of the holder <NUM> when the bed <NUM> is held by the holder <NUM>, so the case <NUM> can be stably held and fixed on the holder <NUM> by interference of the holder <NUM> and the wings <NUM>.

Further, a label (not shown) may be attached to sides of the wings <NUM> or the sides may be used as spaces for writing texts. Accordingly, the analyte collecting apparatus <NUM> can be systemically managed.

The piston <NUM> may be provided to be inserted into the internal space <NUM> through the opening formed at the case <NUM> and may be provided to reciprocate in the internal space <NUM>. Further, the piston <NUM> includes at least one partition wall <NUM> dividing the internal space <NUM>. Further, the piston <NUM> may further include a center column <NUM>, a piston head <NUM>, a flange <NUM> and a sealing member <NUM>.

The center column <NUM>, for example, may be provided in a cylindrical shape and is provided to connect the piston head <NUM> and the partition wall <NUM>. Further, the column <NUM> may connect a plurality of partition walls <NUM>, and the portion connecting the piston head <NUM> and the partition wall <NUM> and the portion connecting the plurality of partition walls <NUM> may be different in thickness. For example, the thickness of the portion connecting the piston head <NUM> and the partition wall <NUM> may be set larger than the thickness of the portion connecting the plurality of partition walls <NUM>, and accordingly, the column <NUM> may occupy the spaces of the sections 102a, 102b, 102c, and 102d in a limitative manner. However, this is only an example, and the center column <NUM> may have entirely the same thickness, or the thickness of the portion connecting the plurality of partition walls <NUM> may be set larger than the thickness of the portion connecting the piston head <NUM> and the partition wall <NUM>.

The piston head <NUM> may be connected to an end of the center column <NUM>, and may be provided to be selectively clamped by a clamp <NUM> of the plunger <NUM> to be described below. Further, the piston head <NUM> may be formed in a disc shape having a radius larger than the center column <NUM>, and may be provided as a flange shape for the center column <NUM>.

The partition walls <NUM> may radially extend from the circumferential surface of the center column <NUM> to be spaced apart from each other. Further, fourth partition walls <NUM> may be provided and can divide the internal space into a total of four sections 102a, 102b, 102c, and 102d, but the present disclosure is not limited thereto, and a certain number of partition walls may be provided, if necessary.

For example, referring to <FIG>, an analyte collecting device <NUM>' according to another embodiment of the present disclosure is proposed. In this embodiment, unlike the embodiment described above, the number of partition walls <NUM> may be two, and accordingly, the number of sections 102a and 102b may be two. According to the analyte collecting device <NUM>' having this configuration, only processing by one solution may be performed in the internal space <NUM> and a plurality of devices may be required for processing that requires a plurality of stages. However, there is the advantage that the size of the devices is correspondingly decreased, the manufacturing cost is reduced, and the devices can be easily used when only one processing is required. However, the present disclosure is not limited to the numbers of the partition walls and the sections, and three or more partition walls and sections may be formed, depending on cases.

Referring to <FIG> again, the flange <NUM> may be attached to at least one surface of two surfaces of the partition wall <NUM> and may be provided such that the circumferential surface is closer to the inner wall forming the internal space <NUM> of the case <NUM> than the circumferential surface of the partition wall <NUM>. For example, when the flange <NUM> and the partition wall are provided in circular disc shapes, the radius of the flange <NUM> may be larger than the radius of the partition wall <NUM>. Due to this shape of the flange <NUM>, when two flanges <NUM> are attached to two sides of the partition wall <NUM>, respectively, a space may be formed along the circumference of the partition wall <NUM> between the two flanges <NUM>. The sealing member <NUM> may be provided in the space formed in this way.

The sealing member <NUM> may surround the circumferential surface of the partition wall <NUM> and may be in contact with the inner wall of the case <NUM>. The sealing member may be an O-ring made of a material such as rubber. By the sealing member <NUM>, the gap between the partition wall <NUM> and the inner wall of the case <NUM> can be sealed by the sealing member <NUM> and the substances accommodated in the sections 102a, 102b, 102c, and 102d can be prevented from leaking from the corresponding sections. Further, even though the piston <NUM> is moved in the internal space <NUM>, the sealing member <NUM> may be interfered with by the flange <NUM>, so that the sealing member <NUM> can keep positioned between the two flanges <NUM> without separating from the circumferential surface of the partition wall <NUM>.

A process of collecting a sample as an analyte through a process such as refinement and a process of performing a predetermined examination on the collected analyte can be manually performed by the analyte collecting device <NUM> having the configuration described above, and may be automatically performed by a system <NUM> for examining an analyte. When the system <NUM> for examining an analyte is used, precise control is possible, as compared with manual work, so collection and examination of an analyte can be more systematically performed. Hereafter, the system <NUM> for examining an analyte according to an embodiment of the present disclosure is described.

Referring to <FIG>, the system <NUM> for examining an analyte according to an embodiment of the present disclosure may be configured to not only perform a predetermined examination on an analyte collected through the analyte collecting device <NUM>, but perform processing, such as refinement, and collection on the analyte. However, depending on cases, the system <NUM> for examining an analyte may also be configured to perform only processing, such as refinement, and collection on an analyte.

The system <NUM> for examining an analyte may include a holder <NUM>, an aggregating device <NUM>, a de-aggregating device <NUM>, a plunger <NUM>, and a controller <NUM>.

The holder <NUM> may be provided to separably hold the analyte collecting device <NUM>. Further, the top of the holder <NUM> may have a shape that can be fastened to the bed <NUM> of the analyte collecting device <NUM>. The holder <NUM> may have a width and a shape that can be inserted in the space between two wings <NUM>. Accordingly, a process for collecting an analyte of the analyte collecting device <NUM> may be performed with the analyte collecting device <NUM> mounted by the holder <NUM>.

The aggregating device <NUM> may be provided to selectively apply a magnetic force toward the aggregating groove <NUM> of the analyte collecting device <NUM> so that the magnetic substance in a solution accommodated in the internal space <NUM> and a substance binding to the magnetic substance can be aggregated in the aggregating groove <NUM>. The aggregating device <NUM> may be controlled by the controller <NUM> to be described below and may be provided as a member such as an electromagnet that can be supplied with power and can generate a magnetic force. Further, the aggregating device <NUM> may be provided inside the holder <NUM>, and may be disposed at a position close to the aggregating groove <NUM> when the analyte collecting device <NUM> is mounted in the holder <NUM>.

The de-aggregating device <NUM> may be provided to selectively apply a magnetic force the internal space <NUM>. The de-aggregating device <NUM> may be controlled by the controller <NUM> and may be provided as a member such as an electromagnet that can be supplied with power and can generate a magnetic force. Further, the de-aggregating device <NUM> may be disposed adjacent to a side of the top of the case <NUM>, and may be operated to apply a magnetic force to the internal space <NUM> of the case <NUM> using power supplied from the outside. Accordingly, the controller <NUM> controls the de-aggregating device <NUM> with a magnetic substance aggregated in the aggregating groove <NUM>, whereby the magnetic substance aggregated in the aggregating groove <NUM> and the substance bound to the magnetic substance can be separated by the magnetic force applied to the internal space <NUM>, and accordingly, the substance binding to the magnetic substance can float back into the internal space <NUM>.

The plunger <NUM> may translate the piston <NUM> in the internal space <NUM> by pushing or pulling the piston head <NUM>. To this end, the plunger <NUM> may include a clamp <NUM> that can selectively hold the piston head <NUM>. The clamp <NUM> may have a shape corresponding to the shape of the piston head <NUM>, and may be provided to selectively hold the piston head <NUM>.

The controller <NUM> may be provided to control the components of the system <NUM> for examining an analyte. In detail, the controller <NUM> may be provided to control at least one or more of the holder <NUM>, the aggregating device <NUM>, the de-aggregating device <NUM>, and the plunger <NUM>. The controller <NUM>, for example, may be a small built-in computer and may include programs, a memory, and a CPU that is a data processing unit. The programs may include an algorithm for controlling at least one or more of the holder <NUM>, the aggregating device <NUM>, the de-aggregating device <NUM>, and the plunger <NUM>. Further, the programs may be stored in a computer memory medium, for example, a memory such as a flexible disc, a compact disc, a hard disc, and a magneto-optical disk (MO), and may be installed in the controller <NUM>. For example, predetermined processing that is performed in the analyte collecting device <NUM> by the system <NUM> for examining an analyte may include a plurality of stages, and, as the controller <NUM> controls the operation of the plunger <NUM>, the plurality of stages can be sequentially performed.

Hereafter, the plurality of stages that is performed by the analyte collecting device <NUM> and the system <NUM> for examining an analyte is described with reference to <FIG>. In the following description, although it is exemplified that an analyte that is collected by the analyte collecting device <NUM> is a refined nucleic acid and includes a specimen for Polymerase Chain Reaction (PCR), the present disclosure is not limited thereto, and the analyte collecting device and the system for examining an analyte may be used to collect other kinds of analytes.

First, a sample can be put into the internal space <NUM> of the analyte collecting device <NUM> (see, <FIG>). The sample that is put into the internal space <NUM> may be liquid, solid, or a mixture including some or all of a cell, a virus, a tissue, exosome, protein, nucleic acid, an antigen, and an antibody, and for example, may be a specimen taken from a human body.

Next, the piston <NUM> is moved in the internal space <NUM> and predetermined processing including a plurality of stages can be sequentially performed. The stage in which predetermined processing is sequentially performed is described through an example as follows. First, a biomaterial contained in a sample is dissolved by the solution in the first section 102a and nucleic acid in the biomaterial can be bound to a magnetic substance (see, <FIG>). Thereafter, the controller <NUM> applies a magnetic force to the aggregating groove <NUM> by controlling the aggregating device <NUM>, and the magnetic substance bound to the nucleic acid can be aggregated in the aggregating groove <NUM> (see, <FIG>). After the magnetic substance bound to the nucleic acid is aggregated in the aggregating groove <NUM>, the controller <NUM> pushes backward the piston <NUM> such that the second section 102b is disposed over the aggregating groove <NUM> (see, <FIG>).

After finishing moving the piston <NUM>, the controller <NUM> can remove the magnetic force applied to the aggregating groove <NUM> by stopping operation of the aggregating device <NUM> and can apply a magnetic force to the internal space <NUM> by operating the de-aggregating device <NUM> (see, <FIG>). Accordingly, the nucleic acid and the magnetic substance bound to each other in the aggregating groove <NUM> can be separated, and the magnetic substance bound to the nucleic acid can float into the internal space of the case. Thereafter, the nucleic acid bound to the magnetic substance can be started to be refined by the solution in the second section 102b.

After a predetermined time for which the nucleic acid can finish being refined passes, the controller <NUM> can aggregate the nucleic acid bound to the magnetic substance in the aggregating groove <NUM> by applying a magnetic force to the aggregating groove <NUM> by operating the aggregating device <NUM> again (see, <FIG>). Thereafter, the controller <NUM> moves backward the piston <NUM> by operating the plunger <NUM>, whereby the third section 102c can be disposed over the aggregating groove <NUM> (see, <FIG>).

After finishing moving the piston <NUM>, the controller <NUM> can remove the magnetic force applied to the aggregating groove <NUM> by stopping operation of the aggregating device <NUM> and can apply a magnetic force to the internal space <NUM> by operating the de-aggregating device <NUM> (see, <FIG>). Accordingly, the nucleic acid and the magnetic substance bound to each other in the aggregating groove <NUM> can be separated, and the magnetic substance bound to the nucleic acid can float into the internal space of the case <NUM>. Thereafter, a process in which the nucleic acid bound to the magnetic substance is eluted from the magnetic substance by the solution in the third section 102c can be performed.

After a predetermined time for which the eluting of the nucleic acid is completed passes, the controller <NUM> can aggregate the magnetic substance separated from the nucleic acid in the aggregating groove <NUM> by applying a magnetic force to the aggregating groove <NUM> by operating the aggregating device <NUM> again (see, <FIG>). Thereafter, the controller may allow the piston <NUM> to be pushed toward the end of the internal space <NUM> in the internal space <NUM>, thereby pushing the third section 102c toward the blowback portion <NUM> containing the eluted nucleic acid. Further, the nucleic acid pressed toward the blowback <NUM> can be discharged as an analyte through the exhaust port <NUM> formed at the case <NUM> (see, <FIG>). In this case, the collecting container X may have been fastened to the container coupling protrusion <NUM>, and the analyte discharged through the exhaust port <NUM> can be collected in the collecting container X and used for a predetermined examination procedure such as PCR.

Meanwhile, the system <NUM> for examining an analyte, as shown in <FIG>, may be configured such that a single system is controlled by one controller <NUM>, but, as shown in <FIG>, a plurality of systems may be controlled. In other words, as in the analyte collecting device <NUM>' shown in <FIG>, when only two sections are provided and one section is filled with a solution for one processing, a plurality of analyte collecting devices <NUM> may be required to perform processing that should be performed through a plurality of stages. As described above, when it is required to operate a plurality of analyte collecting devices in one process, a system <NUM>' for examining an analyte shown in <FIG> may be used.

Hereafter, the system <NUM>' for examining an analyte according to another embodiment of the present disclosure is described.

Referring to <FIG>, the system <NUM>' for examining an analyte may include a plurality of analyte collecting devices <NUM> and may include holders <NUM>, aggregating devices <NUM>, de-aggregating devices <NUM>, a plunger <NUM> to correspond to the number of the analyte collecting devices <NUM>, and may further include conveying lines <NUM> connecting the analyte collecting devices <NUM>, respectively. In this case, the conveying line <NUM> may be provided to connect the exhaust port <NUM> of any one analyte collecting device <NUM> and the injection hole <NUM> of another analyte collecting device <NUM>. Accordingly, an analyte processed through the analyte collecting device <NUM> at the front end is injected into the analyte collecting device <NUM> at the rear end through the conveying line <NUM>, and following processing can be performed.

Further, although not shown in this embodiment, the conveying line <NUM> may further have members such as a pump and a valve for conveying fluid. Further, the conveying line <NUM> may be separably fastened to the analyte collecting device <NUM>, whereby the conveying line <NUM> can be fastened to the analyte collecting device <NUM> or the position thereof can be changed, if necessary. However, this is only an example, and the conveying line <NUM> may be omitted and the analyte discharged from the analyte collecting device <NUM> at the front end may be manually conveyed to the analyte collecting device <NUM> at the rear end.

The controller <NUM> may include a plunger module <NUM>, a de-aggregating module <NUM>, and an aggregating module <NUM>. The plunger module <NUM> may be connected to the plungers <NUM> to independently control the plungers <NUM>. Further, the de-aggregating module <NUM> may be connected to the de-aggregating devices <NUM> to independently control the de-aggregating devices <NUM>. Further, the aggregating module <NUM> may be connected to the aggregating devices <NUM> to independently control the aggregating devices <NUM>. The plunger module <NUM>, the de-aggregating module <NUM>, and the aggregating module <NUM> may be provided as independent algorithms stored in a chipset module or one chipset disposed in the controller <NUM>.

According to the system <NUM>' for examining an analyte described above, a plurality of systems can be independently controlled, so there is the advantage that the system <NUM>' can be especially and usefully used when a process should be performed through multiple stages.

Meanwhile, the configuration of the controller <NUM> of the system for examining an analyte may be changed in various ways. For example, the configuration shown in <FIG> is also possible. Hereafter, the system <NUM>" for examining an analyte according to another embodiment of the present disclosure is described with reference to <FIG>. In the following description, the system <NUM>" for examining an analyte has a difference in the configuration of the controller <NUM>, as compared with the system <NUM>' for examining an analyte described above, so the difference is mainly described and the system <NUM>' for examining an analyte described above is referred to for the same configuration.

Referring to <FIG>, the controller <NUM> of the system <NUM>" for examining an analyte may have a configuration in which modules correspond to systems, respectively, each of which includes one analyte collecting device <NUM>, one holder <NUM>, one aggregating device <NUM>, one de-aggregating device <NUM>, and one plunger <NUM>. For example, when a total of three systems each including one analyte collecting device <NUM>, one holder <NUM>, one aggregating device <NUM>, one de-aggregating device <NUM>, and one plunger <NUM> are provided, the controller <NUM> may include a first module <NUM>, a second module <NUM>, and a third module <NUM> that independently control the systems. Accordingly, there is the advantage that it is possible to easily change the number of low-class systems included in the entire system by removing some of the modules in the controller <NUM> or adding a new module.

Hereafter, the operation and effects of the analyte collecting devices <NUM> and <NUM>' and the systems <NUM>, <NUM>', and <NUM>" for examining an analyte having the configurations described above in accordance with embodiments of the present disclosure are described. The applicant of the present disclosure performed examinations in accordance with an experimental example to be described below to prove the effects of the present disclosure.

Referring to <FIG> and <FIG>, in this experimental example, an analyte collecting device including a first section 102a, a second section 102b, a third section 102c, and a fourth section 102d was prepared, the capacities of the first to third sections 102a, 102b, and 102c were set as <NUM>, <NUM>, and <NUM>µL, and the examination was performed in this state. In this experimental example, Influenza A H1N1 (Human, strain: KUMC-<NUM>) cDNA was conveyed and PCR of the conveyed resultant was performed using the analyte collecting devices according to the embodiments of the present disclosure, and then the result was measured.

The equipment used for PCR was QuantStudio™ <NUM> by ThermoFisher Scientific (Applied Biosystems), and the reagent was PowerUp™ SYBR™ Green Master Mix by the same company. The used primer information is as follows.

PCR reaction volume was <NUM>µL, PowerUp™ SYBR™ Green Master Mix was <NUM>µL, the primer was <NUM>µL, and a reagent of <NUM>µL was included for each experiment.

The PCR procedure was composed of a total of three states. First, a hold stage was performed under the condition of <NUM>/<NUM>. and <NUM>/<NUM>, and a cycling stage was performed for <NUM> cycles under the condition of <NUM>/<NUM> sec. and <NUM>/<NUM> sec. Finally, in a melt curve stage, a result was measured while temperature was increased by <NUM> per <NUM> seconds after <NUM>/<NUM> sec. and <NUM>/<NUM>.

Influenza A H1N1 (Human, strain: KUMC-<NUM>) cDNA that is a sample was produced by QuantiNova™ Reverse Transcription Kit by Qiagen.

In the first section 102a, a solution and magnetic particles required for cell dissolution (lysis) and binding of nucleic acid and magnetic particles were injected, and additionally, Dynabeads® MyOne™ Silane (ThermoFisher Scientific, <NUM>/mL) of <NUM>µL, Lysis/binding Buffer of <NUM>µL by Dynabeads® SILANE viral NA kit (ThermoFisher Scientific), <NUM>-Propanol (Sigma-Aldrich, I9516-<NUM>) of <NUM>µL, Distilled water (DW) of <NUM>µL were injected.

In the second section 102b, a solution for washing was injected in advance, and Washing Buffer2 of <NUM>µL by Dynabeads® SILANE viral NA kit (ThermoFisher Scientific) was injected.

In the third section 102c, a solution for nucleic acid elution was injected in advance, and in the cartridge; Elution Buffer of <NUM>µL and DW of <NUM>µL by Dynabeads® SILANE viral NA kit (ThermoFisher Scientific) were additionally injected in the E. experiment shown in <FIG> and <FIG>, and DW of <NUM>µL was injected in the cartridge, H<NUM>O experiment shown in <FIG> and <FIG>.

In the first section 102a of the analyte collecting device set as described above, Influenza A H1N1 cDNA of <NUM>µL was injected as a sample. Thereafter, analytes were collected in accordance with the methods of collecting an analyte according to the embodiments described above, and PCR was performed by injecting an undiluted solution, a <NUM>/<NUM> diluted solution, and <NUM>/<NUM> diluted solution of <NUM>µL of the collected analytes were injected in individual PCR containers.

As a contrastive group, the same specimen was made pass through the same solutions as the solutions injected in the first section 102a and the second section 102b, and finally, pass through the solution in which Elution Buffer of <NUM>µL and DW of <NUM>µL were injected. Further, an examiner manually performed the experiment using a pipette (pipetting, E. of <FIG> and <FIG>). Similarly, in the contrastive group, PCR was performed by injecting an undiluted solution, a <NUM>/<NUM> diluted solution, and <NUM>/<NUM> diluted solution of <NUM>µL of an analyte in individual PCR containers.

As a standard substances in PCR, <NUM>/<NUM>, <NUM>/<NUM>, and <NUM>/<NUM> diluted solutions (Inf. cDNA of <FIG> and <FIG>) of <NUM>µL of Influenza A H1N1 cDNA that is the same solution as that injected in each experiment were injected in individual PCR containers, whereby PCR was performed.

Since Influenza A H1N1 cDNA injected in each experiment was <NUM>µL and the capacity of the third section 102c was <NUM>µL, when an analyte was <NUM>% conveyed, it corresponds to a <NUM>/<NUM> diluted solution of the standard.

Referring to <FIG>, it was found that when Influenza A H1N1 cDNA solution conveyed using the analyte collecting devices, systems, and methods according to embodiments of the present disclosure was amplified, the threshold cycle (Ct) values measured in the cartridge, E. experiment and the cartridge, H<NUM>O experiment were both similar to the Ct values of the contrastive group of this experimental example and the <NUM>/<NUM> diluted solution of the standard, and the diluted solutions showed the same tendency. The Ct values prove the contrary of the initial cDNA before amplification, so it was determined that collection of an analyte through the analyte collecting devices, systems, and methods according to embodiments of the present disclosure was effective.

Further, referring to <FIG> and <FIG>, it was found that the experiment of injecting distilled water in the third section 102c while using the analyte collecting devices, systems, and methods according to embodiments of the present disclosure has the most similar Ct value to the <NUM>/<NUM> of the standard and the initial concentrations before amplification were similar.

According to the analyte collecting devices and systems for examining an analyte having the configurations described above in accordance with embodiments of the present disclosure, there is an effect that the structures of the devices and the systems are simple, so the cost is low and the devices and the systems can be implemented in a small size. Further, a sample can be effectively processed through an automated process and it is possible to achieve a constant yield ratio of an analyte every time regardless of the skill of the users.

Claim 1:
An analyte collecting device comprising:
a case (<NUM>) including an opening and an internal space (<NUM>); and
a piston (<NUM>) including one or more partition walls (<NUM>) dividing the internal space (<NUM>) into a plurality of internal spaces (102a-d), the piston (<NUM>) being inserted into the internal space (<NUM>) through the opening of the case (<NUM>) to reciprocate in the internal space (<NUM>), characterized in that the case (<NUM>) includes:
an exhaust port (<NUM>) through which the internal space (<NUM>) communicates with an outside and configured such that a sample put in the internal space (<NUM>) can be discharged through the exhaust port (<NUM>) to the outside of the case (<NUM>); and
a specimen injection port (<NUM>) including an injection hole (<NUM>) formed at the case (<NUM>) to allow the internal space (<NUM>) to communicate the outside of the case (<NUM>) and configured such that a sample can be injected.