KR102418920B1 - Syringe device, device and method for extracting nucleic acid using the same - Google Patents

Abstract

A syringe device is provided. The syringe device according to an embodiment of the present invention includes a cylinder body having a fluid outlet on the front side and a piston installed on the rear side; A first packing member that divides the inner space of the cylinder body into a first space adjacent to the fluid outlet and a second space adjacent to the piston rather than the first space, and which is movable forward and backward in the inner space of the cylinder body. and a hole may be formed in the first packing member so that the fluid located in the second space can be moved to the first space according to a pressure difference between the first space and the second space.

Description

Syringe device and nucleic acid extraction device and method using the same

The present invention relates to a syringe device, and more particularly, to a syringe device for sequentially discharging a plurality of contents accommodated therein, and an apparatus and method for extracting nucleic acids from cells using the syringe device.

A method currently used to detect microorganisms, etc. present in food is a method of confirming the genes of microorganisms. The detection method through gene identification has the advantage of being able to accurately determine the type and number of microorganisms.

However, in order to confirm the gene of the microorganism, the process of extracting the nucleic acid from the microorganism is inevitably accompanied.

A general method used to extract nucleic acids with microorganisms is to use nucleic acid extraction equipment such as centrifuges. However, such nucleic acid extraction equipment has a limitation in that it is difficult to manufacture as a small equipment.

Therefore, in order for the experimenter to extract the nucleic acid, there is a time and place limitation in which the nucleic acid must be extracted through the nucleic acid extraction equipment. In particular, there is a problem in that the nucleic acid cannot be immediately extracted from the site where the detection of microorganisms is required.

Recently, various buffers, ie, buffers, are used to easily extract nucleic acids in the field, but there is a problem in that the experimenter must always have various types of buffers and equipment such as a dropper to transfer them.

In addition, in order to extract nucleic acids, the amount of buffer must be adjusted and used, but there is a problem in that the accuracy of nucleic acid extraction varies depending on the skill of the user.

Laid-Open Patent Publication No. 1999-0022612 (DNA extraction method and extraction apparatus (DNA extraction method and extraction apparatus)

An object of the present invention is to provide a syringe device capable of storing and transporting several types of fluids in one syringe, and discharging each fluid step by step.

Another object of the present invention is to provide a syringe device capable of dispensing the same amount of contents with one syringe.

Another object of the present invention is to provide an apparatus and method capable of extracting nucleic acids from cells conveniently in the field by discharging the buffer solution step by step.

In particular, it is an object of the present invention to provide an apparatus and method capable of extracting nucleic acids through simple manipulations even if the user is not skilled.

The problems of the present invention are not limited to the problems mentioned above, and other problems not mentioned will be clearly understood by those skilled in the art from the description below.

In order to solve the above problems, the injector device according to an aspect of the present invention includes a cylinder body having a fluid outlet on the front side and a piston installed on the rear side; A first packing member that divides the inner space of the cylinder body into a first space adjacent to the fluid outlet and a second space adjacent to the piston rather than the first space, and which is movable forward and backward in the inner space of the cylinder body. and a hole may be formed in the first packing member so that the fluid located in the second space can be moved to the first space according to a pressure difference between the first space and the second space.

In this case, further comprising a second packing member installed between the first packing member and the piston, wherein the second packing member moves into the second space and a third space adjacent to the piston rather than the second space. A fluid that partitions the internal space of the body, is movable forward and backward in the internal space of the cylinder body, and is located in the third space according to the pressure difference between the second space and the third space in the second packing member. A hole may be formed so that it can move into the second space.

At this time, the first packing member and the second packing member are each formed of a rubber material, doedoe formed around the central portion and the central portion of the disk shape having an area smaller than the inner space of the cylinder body, the inner space of the cylinder body A ring-shaped rim portion in close contact with the rim portion may be provided, and the rim portion may have an inclined surface elastically supporting the central portion so that the central portion can be moved forward or rearward based on the rim portion.

At this time, the piston is fitted in the inner space of the cylinder body, a piston body movable forward or backward; Doedoe in close contact with the inside of the cylinder body may include a gasket coupled to the front end of the piston body.

At this time, the cylinder body has a first screw thread formed toward the inner side of the cylinder body at the rear portion of the cylinder body, and the piston has a second screw thread formed on the piston body to be screw-engaged with the first screw thread. ; and a handle formed at the other end of the piston body.

In the syringe device according to another aspect of the present invention, the cylinder body is coupled therein, the rear surface is opened, and a hole is formed so that the front end of the cylinder body protrudes to the outside in the front surface in a state in which the cylinder body is coupled, , including a cylinder case having a third screw thread on the outside, and a cover part having a fourth screw thread therein so that a part of the rear part of the cylinder case is inserted therein so as to be screw-coupled with the third screw thread And, the piston is supported on the inside of the cover portion may include a support member formed at the other end of the piston body so that the piston is pressed forward according to the rotation of the cover portion.

In this case, the cylinder case may be formed with a groove in the longitudinal direction so as to confirm a position in which the gasket moves in the cylinder body.

In the syringe device according to another aspect of the present invention, one end of the upper plate and the lower plate is connected while the upper plate and the lower plate maintain a predetermined distance so that a flange coupling groove is formed therebetween, and the cylinder body is fitted at the other end. and a piston fixing member having an opening formed in the upper plate and the lower plate so that the cylinder body includes a cylinder flange extending outwardly around the rear end of the cylinder body so as to be fitted into the flange coupling groove and, the piston body includes a plurality of piston fixing grooves formed in the piston body at predetermined intervals in the longitudinal direction, and the piston fixing member is coupled to the plurality of piston fixing grooves to support the piston body. fixed part; Connecting the piston fixing part to one end of the upper plate and the lower plate, but may include a bending part elastically supporting the piston fixing part.

In this case, a filter member connectable to the cylinder body to filter the fluid passing through the fluid outlet is included, and the filter member is disposed in front of the cylinder body so that the filter member is disposed on the front surface of the fluid outlet. It may be coupled to a filter support member that may be coupled.

Nucleic acid extraction apparatus using a syringe device according to an aspect of the present invention, a cylinder body having a fluid outlet on the front side and a piston installed on the rear side; a first packing member partitioning the inner space of the cylinder body into a first space adjacent to the fluid outlet and a second space adjacent to the piston rather than the first space, the first packing member being movable forward and backward in the inner space of the cylinder body; It is installed between the first packing member and the piston to partition the inner space of the cylinder body into the second space and a third space adjacent to the piston rather than the second space, to the front and rear sides within the inner space of the cylinder body A movable second packing member is included, and the first packing member has a hole in the first packing member so that the fluid located in the second space can be moved to the first space according to the pressure difference between the first space and the second space. is formed, and the second packing member has a hole formed therein so that the fluid located in the third space can be moved to the second space according to the pressure difference between the second space and the third space, using a syringe device A nucleic acid extraction device comprising: a filter member formed to filter a cell lysate containing nucleic acids in the first space when discharged to the fluid outlet to leave the nucleic acids and cellular foreign substances in the first space; and a filter support member coupled to the front of the cylinder body while being coupled to the filter member so that the filter member is disposed on the front surface of the fluid outlet, wherein the second space has a first wash buffer as a wash buffer. A solution may be accommodated, and a second solution may be accommodated as an elution buffer in the third space.

In the nucleic acid extraction method using the nucleic acid extraction apparatus according to an aspect of the present invention, the cell lysate is introduced into the first space by separating the filter member and reversing the piston to introduce the cell lysate into the first space through the fluid outlet. step; a cell lysate filtering step of coupling the filter member to the fluid outlet, and discharging the cell lysate accommodated in the first space by energizing the piston; a first solution discharging step of advancing the piston to discharge the first solution accommodated in the second space through the first space through the fluid outlet; a second solution discharging step of advancing the piston to discharge the second solution accommodated in the third space into the first space through the second space; and discharging the nucleic acid solution accommodated in the first space by advancing the piston through the fluid outlet.

The syringe device according to an aspect of the present invention can store and transport several types of fluids with one syringe by having a packing member having a hole formed therein, and discharge each fluid step by step.

The syringe device according to an aspect of the present invention can dispense the same amount of contents with one syringe by having a packing member having a hole formed therein.

The nucleic acid extraction apparatus and method using a syringe device according to an aspect of the present invention can extract a nucleic acid from a cell conveniently in the field by discharging the buffer step by step by having a packing member having a hole formed therein.

The apparatus and method for extracting nucleic acids using a syringe device according to an aspect of the present invention can extract nucleic acids easily even if a user using screw coupling is not skilled.

1 is a perspective view of a nucleic acid extraction apparatus according to a first embodiment of the present invention.
2 is a cross-sectional view of the nucleic acid extraction apparatus according to the first embodiment of the present invention.
3 (a) to (c) are enlarged cross-sectional views of the first packing member of the nucleic acid extraction apparatus according to the first embodiment of the present invention.
Figure 4 (a) is a view showing the preparation state of the cell lysate inflow of the nucleic acid extraction apparatus according to the first embodiment of the present invention.
Figure 4 (b) is a view showing the state of the cell lysate inflow of the nucleic acid extraction apparatus according to the first embodiment of the present invention.
Figure 5 (a) is a view showing a state in which the filter member of the nucleic acid extraction apparatus according to the first embodiment of the present invention is coupled.
Figure 5 (b) is a view showing a filtered state of the cell lysate of the nucleic acid extraction apparatus according to the first embodiment of the present invention.
Figure 6 (a) is a view showing a state in which the first solution of the nucleic acid extraction apparatus according to the first embodiment of the present invention is discharged.
6 (b) is a view showing a state in which the second solution of the nucleic acid extraction apparatus according to the first embodiment of the present invention is discharged.
Figure 6 (c) is a view showing a state in which the nucleic acid lysate of the nucleic acid extraction apparatus according to the first embodiment of the present invention is discharged.
7 is a flowchart illustrating a method for extracting nucleic acids using the nucleic acid extraction apparatus according to the first embodiment of the present invention.
8 is a cross-sectional view of a nucleic acid extraction apparatus according to a second embodiment of the present invention.
9 is an enlarged perspective view of the cylinder case and the cover of the nucleic acid extraction apparatus according to the second embodiment of the present invention.
10 is a cross-sectional view of a nucleic acid extraction apparatus according to a third embodiment of the present invention.
11 is an enlarged perspective view of the piston fixing member of the nucleic acid extraction apparatus according to the third embodiment of the present invention.

Hereinafter, with reference to the accompanying drawings, embodiments of the present invention will be described in detail so that those of ordinary skill in the art can easily carry out the present invention. The present invention may be embodied in many different forms and is not limited to the embodiments described herein. In order to clearly explain the present invention in the drawings, parts irrelevant to the description are omitted, and the same reference numerals are given to the same or similar components throughout the specification.

The syringe device according to an aspect of the present invention is a device for sequentially discharging the contents accommodated in the syringe inner space partitioned by the packing member. Such a syringe device may be provided for a dispensing device for dispensing and dispensing the contents in the same amount, an extraction device for extracting nucleic acids or extracellular vesicles by discharging the buffer in stages, and the like. In the present specification, for convenience of description, it is assumed that the syringe device is implemented as a device for extracting nucleic acids, and the nucleic acid extraction device according to various embodiments of the present invention will be described.

1 is a perspective view of a nucleic acid extraction apparatus according to a first embodiment of the present invention. 2 is a cross-sectional view of the nucleic acid extraction apparatus according to the first embodiment of the present invention. 3 (a) to (c) are enlarged cross-sectional views of the first packing member of the nucleic acid extraction apparatus according to the first embodiment of the present invention.

In the following description, as seen in FIG. 1, the fluid outlet side of the nucleic acid extraction device is defined as the front, the piston side is defined as the rear, and the direction extending from the front to the rear is defined as the longitudinal direction.

1 and 2 , the nucleic acid extraction apparatus 1 according to the first embodiment of the present invention includes a cylinder body 10 , a first packing member 40 and a piston 30 .

The cylinder body 10 is formed to extend to the rear side so that a space in which the fluid can be accommodated is formed in the cylinder body 10 . At this time, the cross-section of the cylinder body 10 is preferably formed in a circular shape to facilitate sealing the fluid accommodated in the inner space of the cylinder body 10 . However, if a space is formed inside the cylinder body 10 to accommodate the fluid, the shape of the cross-section of the cylinder body 10 may be variously formed.

At this time, the fluid outlet 12 is formed on the front side of the cylinder body so that the contents accommodated in the cylinder body 10 can communicate with the outside in fluid.

At this time, the fluid outlet 12 is formed to protrude from the front end of the cylinder body 10 to the front side. In addition, the fluid outlet 12 is formed to pass through the inside and the outside of the cylinder body 10 as a fluid communication channel.

At this time, the cross-sectional area of the flow path formed in the fluid outlet 12 is the cylinder body 10 so that the fluid accommodated in the cylinder body 10 is discharged to the outside through the fluid outlet 12 to receive a high pressure. ) is formed relatively smaller than the cross-sectional area of the space formed inside.

In addition, the front portion of the cylinder body 10 is formed such that the cross-sectional area of the cylinder body 10 becomes narrower toward the fluid outlet 12 so that the fluid discharged from the cylinder body 10 is concentrated to the fluid outlet 12 and the fluid It is preferable that it is formed so that it can receive a higher pressure when it is discharged to the outside.

The rear side of the cylinder body 10 is formed to be opened, and the piston 30 is coupled to the cylinder body 10 through the opening. At this time, the piston 30 is fitted with at least a part inserted into the cylinder body 10 .

When describing the piston 30 in more detail, as shown in FIGS. 1 and 2 , the piston 30 of the nucleic acid extraction device according to the first embodiment of the present invention includes a piston body 32 and a gasket 34 . and a handle 38 .

The piston body 32 is formed longer than the length in the longitudinal direction of the cylinder body 10 so that the front part of the piston body 32 can be moved forward of the cylinder body 10 while being fitted inside the cylinder body 10 . do.

At this time, a gasket 34 is provided at the front end of the piston body 32 . The gasket 34 is in close contact with the inside of the cylinder body 10 to seal the space inside the cylinder body 10 and prevent the fluid accommodated in the cylinder body 10 from flowing out to the rear side.

At this time, the gasket 34 is not limited in material or shape as long as it can seal the inner space of the cylinder body 10 .

On the other hand, as shown in Figures 1 and 2, the piston body 32 is provided with a second screw thread (26). The second thread 26 is screwed into the first thread 36 formed on the inside of the cylinder body 10 . With this screwing, the piston 30 may move forward or backward in the cylinder body 10 according to the rotational direction.

At this time, the first screw thread 36 is formed at the rear end of the cylinder body 10 so as not to interfere with sealing the inside while the gasket 34 is fitted in the cylinder body 10 .

The piston body 32 is provided with a handle 38 at the rear end so that the user can easily rotate the piston. As shown in Fig. 1, the handle 38 is preferably formed with grooves at regular intervals around it so that the user can easily grip it.

Meanwhile, in front of the gasket 34 of the piston 30 , the first packing member 40 is disposed to be in close contact with the inside of the cylinder body 10 .

The first packing member 40 may move forward or rearward in a state in close contact with the inside of the cylinder body 10 according to the movement of the piston 30 . The first packing member 40 is formed of a material, for example, rubber, to which the first packing member 40 can be in close contact with the inside of the cylinder body 10 .

At this time, a space adjacent to the front side of the first packing member 40 partitioned inside the cylinder body 10 by the first packing member 40 , that is, the fluid outlet 12 is defined as the first space 14 . and a space adjacent to the rear side of the first packing member 40, that is, the piston 30, is defined as the second space 16.

A hole 42 is formed in the first packing member 40 to pass through the first space 14 and the second space 16 . When pressure is formed in the first space 14 and the second space 16 , the first packing member 40 is compressed by being pressed to the rear side in the first space 14 and toward the front side in the second space 16 , and the length The hole 42 is tightened while expanding in a direction perpendicular to the direction. Accordingly, the fluid accommodated in the cylinder body 10 cannot flow through the hole 42 formed in the first packing member 40 .

When the user moves the piston 30 forward, the pressure of the second space 16 increases and the first packing member 40 moves forward to maintain the pressure equilibrium state. At this time, when the first packing member 40 reaches the front end inside the cylinder body 10, the pressure of the second space 16 is formed to be greater than the pressure of the first space 14, so that a pressure difference occurs and the first As the packing member 40 is pressed and deformed toward the first space 14 , the hole 42 formed in the first packing member 40 is opened, and the fluid in the second space 16 is discharged through the hole 42 . It can be moved to one space (14).

Hereinafter, a structure of fluid movement through the hole 42 of the first packing member 40 will be described in more detail with reference to FIGS. 3 (a) to (c).

3 (a) to (c), the first packing member 40 is provided with a central portion 44, an edge portion 46, and an inclined surface (48).

As shown in Fig. 3 (a), the central portion 44 is formed in the shape of a disk having an area smaller than the cross-sectional area perpendicular to the longitudinal direction of the cylinder body (10). At this time, the hole 42 formed in the first packing member 40 is formed in the central part 44, and is preferably located in the center of the central part 44 so that the tightening or spreading deformation is easily made.

At this time, as shown in Fig. 3 (a), the periphery of the central portion 44, the edge portion 46 is formed. The edge portion 46 supports the central portion 44 to be vertically disposed in the longitudinal direction inside the cylinder body 10, and is in close contact with the inside of the cylinder body 10 so that the fluid inside the cylinder body 10 is the first packing member It is prevented from leaking from the part except the hole of (40).

At this time, as shown in Fig. 3 (a), the edge portion 46 is inclined surface 48 connecting the circumference of the central portion 44 and the inner circumference of the edge portion 46 in order to support the central portion 44. This may be provided. The inclined surface 48 is formed to form an acute angle with a cross section perpendicular to the longitudinal direction from the edge portion 46 to the rear side, and may be formed at various angles.

At this time, the inclined surface 48 elastically supports the periphery of the central portion 44 so that the central portion 44 is pressed toward the edge portion 46 to be easily deformed.

As the inclined surface is provided, the central portion 44 is located on the rear side of the edge portion 46 . Accordingly, when the user moves the piston 30 forward, as shown in FIG. 3 ( b ), the rim portion 46 reaches the front end of the cylinder body 10 while being higher than the central portion 44 .

At this time, if the user continuously moves the piston 30 forward, as shown in FIG. 3 ( b ), the rim portion 46 is supported by the inner front end of the cylinder body 10 , and the piston 30 ) compresses the fluid accommodated in the second space 16 , thereby increasing the pressure of the second space 16 . Accordingly, the fluid accommodated in the second space 16 intensively presses the central portion 44 .

At this time, as shown in Fig. 3 (c), the central portion 44 elastically supported by the inclined surface 48 is pressed toward the front side, so that the inclined surface 48 is moved from the edge portion 46 to the front side in the longitudinal direction. It is deformed to form an acute angle with a vertical section. That is, the first packing member 40 is deformed such that the central portion 44 moves forward while the rim portion 46 is supported by the front end inside the cylinder body 10 .

As the first packing member 40 is deformed, as shown in Fig. 3 (c), the hole 42 formed in the central portion 44 is opened to the front side, and the first space 14 and the second space ( 16), the fluid moves to the first space 14 due to the pressure difference.

At this time, when the user moves the piston 30 more forward, the fluid accommodated in the first space 14 is discharged to the outside through the fluid outlet 12 .

In addition, as shown in FIG. 2 , the nucleic acid extraction apparatus 1 according to the first embodiment of the present invention may further include a second packing member 41 .

Like the first packing member 40 , the second packing member 41 may move forward or rearward in a state in close contact with the inside of the cylinder body 10 according to the movement of the piston 30 . In addition, the second packing member 41 is also formed of a material that can be in close contact with the inside of the cylinder body 10, for example, rubber.

At this time, the space adjacent to the front side of the second packing member 41 that is partitioned inside the cylinder body 10 by the second packing member 41 , that is, the first packing member 40 is the second space 16 . and defining a space adjacent to the rear side of the second packing member 41 , that is, the piston 30 , as the third space 18 .

A hole 42 is formed in the second packing member 41 to pass through the second space 16 and the third space 18 like the first packing member 40 . When pressure is formed in the second space 16 and the third space 18, the second packing member 41 is compressed by being pressed to the rear side in the second space 16 and to the front side in the third space 18, and the length The hole 42 is tightened while expanding in a direction perpendicular to the direction. Accordingly, the fluid accommodated in the cylinder body 10 cannot flow through the hole 42 formed in the second packing member 41 .

The second packing member 41 is different from the first packing member 40 only in the position disposed in the interior of the cylinder body 10 and the space partitioned accordingly, and other shapes and functions are the same. Accordingly, in the following description of the second packing member 41 , a detailed description of the same configuration and operation as that of the first packing member 40 is omitted and the description of the first packing member 40 is replaced.

When the user moves the piston 30 forward, the second packing member 41 moves forward together with the first packing member 40 . At this time, the internal pressure of the cylinder body 10 is increased. However, since the first space 14 is capable of fluid communication with the outside by the fluid outlet 12 , the pressure is lower than that of the second space 16 and the third space 18 . According to this pressure difference, the first packing member 40 moves forward faster than the second packing member 41 , and the solution accommodated in the second space 16 while reaching the front end of the cylinder body 10 . It is possible to move to the first space 14 before the solution accommodated in the third space 18 .

On the other hand, the nucleic acid extraction apparatus 1 according to the first embodiment of the present invention has been described as having the first packing member 40 and the second packing member 41 for convenience of description, but if necessary, the first packing member In addition to the (40) and the second packing member (41), at least one packing member having the same shape as the first packing member (40) may be further provided. In addition, the cylinder body 10 may be formed to extend in the longitudinal direction according to the number of added packing members.

1 and 2 , the nucleic acid extraction apparatus 1 according to the first embodiment of the present invention may further include a filter member 20 and a filter support member 22 .

The filter member 20 is disposed at the front end of the fluid outlet 12 to filter the fluid discharged from the cylinder body 10 to the fluid outlet 12 .

The filter member 20 has a porous structure to filter out nucleic acids and cellular foreign substances. The size of the hole formed in the filter member 20 may be changed according to the size of the filtering target. At this time, since the filter member 20 having various types of porous structures is known, a detailed description thereof will be omitted.

In this case, the cellular foreign matter filtered by the filter member 20 together with the nucleic acid refers to a composition of the destroyed cells included in the cell lysate except for the nucleic acid to be detected.

The filter member 20 has a mesh support (not shown) on the front side of the filter member 20 so as to filter nucleic acids and cellular foreign substances without being damaged by high pressure generated when the fluid is discharged through the fluid outlet 12 . and on the rear side.

At this time, the filter support member 22 is coupled to the fluid outlet 12 so that the filter member 20 is fixed to the front end of the fluid outlet 12 .

More specifically, the rear end of the filter support member 22 is formed to have a hole having the same area as a cross section perpendicular to the longitudinal direction of the fluid outlet 12 in order to be coupled to the fluid outlet 12 . Accordingly, the front portion of the fluid outlet 12 is coupled to the rear end of the filter support member 22 .

At this time, if the fluid discharged from the fluid outlet 12 through the coupling between the filter support member 22 and the fluid outlet 12 can be discharged through the filter member 20, various coupling methods, for example, It may be coupled by screw coupling, fitting coupling, or the like.

At this time, the inner diameter of the front portion of the filter support member 22 is formed to be smaller than the inner diameter of the rear portion of the filter support member 22 . A filter member 20 is disposed on a jaw formed according to the difference in inner diameter between the front and rear portions of the filter support member 22 . Accordingly, even if the filter member 20 receives an external force toward the front by the fluid discharged from the fluid outlet 12 , the filter member 20 may be fixed to the front end of the fluid outlet 12 .

In addition, the inner diameter of the front portion of the filter support member 22 is preferably formed to be the same as the inner diameter of the fluid outlet (12). Accordingly, the fluid can be discharged without interference due to a difference in cross-sectional area of the flow path from the fluid outlet 12 to the filter support member 22 .

Hereinafter, a method for extracting nucleic acids from the cell lysate 74 using the nucleic acid extraction apparatus 1 according to the first embodiment of the present invention will be described with reference to FIGS. 4 to 7 .

4 (a) and (b) are diagrams showing a cell lysate preparation state and a cell lysate inflow state of the nucleic acid extraction apparatus according to the first embodiment of the present invention, respectively. 5 (a) and (b) are views showing a state in which the filter member of the nucleic acid extraction apparatus according to the first embodiment of the present invention is coupled and a state in which the cell lysate is filtered, respectively. 6 (a) to (c) are views showing a state in which the first solution is discharged, the state in which the second solution is discharged, and the state in which the nucleic acid solution is discharged, respectively, of the nucleic acid extraction apparatus according to the first embodiment of the present invention; to be. 7 is a flowchart illustrating a method for extracting nucleic acids using the nucleic acid extraction apparatus according to the first embodiment of the present invention.

Meanwhile, in the following, the first solution 70 is accommodated in the second space 16 of the nucleic acid extraction apparatus 1 according to the first embodiment of the present invention, and the second solution 72 is accommodated in the third space 18 . A nucleic acid extraction method will be described with the premise that this will be accepted.

At this time, the first solution 70 is a wash buffer for washing the rest except for nucleic acids. Accordingly, cellular foreign substances other than nucleic acids of the cells to be detected filtered by the filter member 20 are dissolved in the first solution 70 .

The wash buffer used as the first solution 70 may be made of various compositions depending on the cell having the nucleic acid to be extracted, and the embodiment is not limited thereto. At this time, since the working principle of the wash buffer is known, a detailed description thereof will be omitted.

The second solution 72 is an elution buffer for dissolving nucleic acids. Accordingly, the nucleic acids of the cells to be detected are dissolved in the second solution 72 , except for foreign substances dissolved and discharged by the first solution 70 .

The solution used as the second solution 72 may be composed of various compositions depending on the cell having the nucleic acid to be extracted, and the embodiment is not limited thereto. At this time, since the working principle of the elution buffer is known, a detailed description thereof will be omitted.

7, the method for extracting nucleic acids using the nucleic acid extraction apparatus according to the first embodiment of the present invention includes a cell lysate inflow step (S10), a cell lysate filtering step (S20), the first It includes a solution discharging step (S30), a second solution discharging step (S40) and a nucleic acid solution discharging step (S50).

At this time, hereinafter, the rotation direction for moving the piston 30 forward is defined as the forward direction, and the rotation direction for moving the piston 30 backward is defined and described as the reverse direction.

As shown in Fig. 4 (a), the user separates the filter support member 22 of the nucleic acid extraction device 1 and inserts the fluid outlet 12 into a separate container in which the target cell lysate 74 is accommodated. insert

When the user rotates the handle 38 of the piston 30 in the reverse direction to move the piston 30 backward in the inserted state, the pressure in the first space 14 is lowered as shown in Fig. 4 (b). The cell lysate 74 is introduced into the first space 14 through the fluid outlet 12 . (S10)

At this time, the user fastens the separated filter support member 22 to the fluid outlet 12 while the cell lysate 74 is accommodated in the first space 14, as shown in FIG. 5 (a). . When the user slowly rotates the handle 38 in the forward direction to advance the piston 30 while the filter support member 22 is fastened, the first space 14 and the second space 16 within the cylinder body 10 And the pressure of the third space 18 is increased.

At this time, the pressure of the first space 14, the second space 16, and the third space 18 moves to the front side so that the first packing member 40 and the second packing member 41 are balanced with each other. While doing this, the cell lysate 74 accommodated in the first space 14 is discharged to the fluid outlet 12 .

At this time, the user can move the piston 30 forward by slowly rotating the piston 30 in the forward direction while looking at the inside of the cylinder body 10 with the naked eye so that the first packing member 40 and the second packing member 41 are not deformed. That is, the user visually determines the first packing member 40 according to the state of the nucleic acid extracting device 1 and the amount of contents accommodated in the first space 14 , the second space 16 and the third space 18 . And while observing the movement of the second packing member 41, the forward speed and the moving distance of the piston 30 can be easily adjusted.

Accordingly, as shown in FIG. 5 (b), the cell lysate 74 passes through the filter member 20 and is discharged to the outside, and is filtered on the cross section of the first space 14 side of the filter member 20. Nucleic acids and cellular debris are located. (S20)

When the user continuously and slowly advances the piston 30, the periphery of the first packing member 40 reaches the front portion of the cylinder body 10, and the first solution 70 accommodated in the second space 16 is 1 The central portion 44 of the packing member 40 is pressed.

At this time, as the central portion 44 of the first packing member 40 moves to the front side, the first packing member 40 is deformed, and a hole 42 formed in the first packing member 40 is shown in FIG. 6 (a). The first solution 70 is discharged into the first space 14 as it opens. (S30)

Accordingly, the filtered cells come into contact with the first solution 70 on the cross section on the side of the first space 14 of the filter member 20 , and cellular foreign substances excluding nucleic acids are dissolved in the first solution 70 .

At this time, when the user advances the piston 30 a little more and discharges the first solution 70 introduced into the first space 14 through the fluid outlet 12, the cellular foreign matter excluding the nucleic acid is removed from the first solution ( 70), the nucleic acid to be extracted is disposed on the cross-section of the first space 14 side of the filter member 20 .

When the user continuously advances the piston 30 slowly, the periphery of the second packing member 41 also reaches the front part of the cylinder body 10 in which the first packing member 40 is located, and enters the third space 18 . The received second solution 72 presses the central portion 44 of the second packing member 41 .

At this time, as the central portion 44 of the second packing member 41 moves forward, the second packing member 41 is deformed, and the hole 42 formed in the second packing member 41 is shown in FIG. 6(b) . The second solution 72 is discharged to the first space 14 through the second space 16 as it opens. (S40)

Accordingly, the filtered nucleic acid is brought into contact with the second solution 72 on the cross section of the first space 14 side of the filter member 20 , and the nucleic acid is dissolved in the second solution 72 .

At this time, when the user advances the piston 30 a little more and discharges the second solution 72 introduced into the first space 14 through the fluid outlet 12 , the nucleic acid is dissolved in the second solution 72 . is discharged, and the user can obtain the target nucleic acid by accommodating the extracted nucleic acid lysate in a separate container. (S50)

As described above, the process of extracting nucleic acids using the nucleic acid extraction apparatus 1 according to the first embodiment of the present invention is simple, and anyone can easily extract nucleic acids using the nucleic acid extraction apparatus 1 without special education or skill. There is an advantage that

Hereinafter, a nucleic acid extraction apparatus 1 ′ according to a second embodiment of the present invention will be described with reference to FIGS. 8 and 9 . However, in the following description of the second embodiment, a detailed description of the same configuration as that of the first embodiment will be omitted, and a configuration different from the first embodiment will be mainly described.

8 is a cross-sectional view of a nucleic acid extracting apparatus according to a second embodiment of the present invention, and FIG. 9 is an enlarged perspective view of a cylinder case and a cover part of the nucleic acid extracting apparatus according to a second embodiment of the present invention.

The nucleic acid extraction device 1 ′ according to the second embodiment of the present invention includes a cylinder body 10 , a first packing member 40 , a second packing member 41 , a piston 30 , a cylinder case 50 and A cover portion (60) is provided.

The inside of the cylinder body 10 of the nucleic acid extracting apparatus 1' according to the second embodiment of the present invention is different from the nucleic acid extracting apparatus 1' according to the first embodiment, the second screw thread 26 is not provided. and the first screw thread 36 is not provided on the piston body 32 .

On the other hand, as shown in FIGS. 8 and 9 , the nucleic acid extraction device 1 ′ according to the second embodiment of the present invention includes a cylinder case 50 into which the cylinder body 10 is fitted. Therefore, the shape of the cross section perpendicular to the longitudinal direction of the cylinder case 50 is the same as the shape of the cross section perpendicular to the longitudinal direction of the cylinder body 10 .

At this time, the cylinder case 50 is formed to extend to the rear by the same length as the length of the cylinder body 10, and the rear surface is opened so that the cylinder body 10 can be fitted.

The front portion of the cylinder case 50 is formed to protrude to the outside so that the user can easily grip it, and the circumference of the front portion may be formed at an angle at regular intervals. At this time, if the user can easily grip it, it may be formed in various shapes, such as grooves formed at regular intervals.

On the other hand, even in a state in which the cylinder body 10 is accommodated in the cylinder case 50, the front surface of the cylinder case 50 so that the fluid outlet 12 formed on the front surface of the cylinder body 10 accommodated therein protrudes to the outside. A hole is formed in the hole, and the fluid outlet 12 is fitted into the hole.

As shown in FIG. 8 , a third screw thread 52 is provided on the outside of the body of the cylinder case 50 . The third screw thread 52 may be formed on the outside of the rear portion of the cylinder case 50, but it is preferable that the third screw thread 52 extends to the front side in order to be stably screwed with the cover part 60 to be described later. .

At this time, a groove 54 is formed in the cylinder case 50 in the longitudinal direction on one side of the body of the cylinder case 50 so that the cylinder body 10 coupled therein is visible to the naked eye.

The user can adjust the piston 30 while visually confirming that the first packing member 40 or the second packing member 41 inside the cylinder body 10 moves through the groove 54 .

On the other hand, the cover portion 60 is formed such that a portion of the rear portion of the cylinder case 50 is coupled therein. At this time, the third screw thread 52 and the fourth screw thread 62 screw-coupled to the inner side of the cover part 60 is formed.

At this time, unlike the nucleic acid extracting apparatus 1' according to the first embodiment, in the nucleic acid extracting apparatus 1' according to the second embodiment of the present invention, according to the rotation of the cover part 60, the piston 30 is A support member 39 is formed at the rear side end of the piston body 32 to move forward.

The support member 39 is in contact with the inside of the rear surface of the cover part 60 , and as the cover part 60 advances, it is pressed to the front side so that the piston 30 advances.

That is, the user screws the cover part 60 with the rear part of the cylinder case 50 while holding the front part of the cylinder case 50 , and then moves the piston 30 forward by rotating the cover part 60 . can do it

In the method of extracting nucleic acids using the nucleic acid extraction apparatus 1' according to the second embodiment of the present invention, the first implementation is performed in the step (S10) of introducing the cell lysate 74 into the first space 14. Unlike the nucleic acid extraction device 1 according to the example, there is a difference in that the user must pull the support member 39 of the piston 30 rearward to introduce the cell lysate 74 into the first space 14 . .

However, the steps other than the cell lysate introduction step (S10) are the same as the process of extracting nucleic acids using the nucleic acid extraction apparatus 1 according to the first embodiment, or at least to the extent that those skilled in the art can easily understand through this. Therefore, detailed description is omitted.

Hereinafter, a nucleic acid extraction apparatus 1" according to a third embodiment of the present invention will be described with reference to FIGS. 10 and 11. However, in the description of the third embodiment, the same A detailed description of the configuration will be omitted, and description will be made focusing on a configuration distinct from the first embodiment.

10 is a cross-sectional view of a nucleic acid extracting apparatus according to a third embodiment of the present invention, and FIG. 11 is an enlarged perspective view of a piston fixing member of the nucleic acid extracting apparatus according to a third embodiment of the present invention.

The nucleic acid extraction device (1") according to the third embodiment of the present invention includes a cylinder body 10, a first packing member 40, a second packing member 41, a piston 30, and a piston fixing member 80. to provide

The piston fixing member 80 is connected with a predetermined interval so that one end of the upper plate 84 and the lower plate 86 having a rectangular shape is formed with a flange coupling groove 89 between the upper plate 84 and the lower plate 86. 11, it is formed in a three-figure shape.

In addition, an opening 85 is formed at the other end of the upper plate 84 and the lower plate 86 of the piston fixing member 80 so that the cylinder body 10 can be fitted.

At this time, as shown in FIG. 10 , the cylinder body 10 is provided with a cylinder flange 13 extending outwardly around the rear end of the cylinder body 10 . The cylinder flange 13 is fitted into the flange coupling groove 89 and disposed between the upper plate 84 and the lower plate 86, and accordingly, the cylinder body 10 is also fitted into the opening 85 and the piston fixing member. (80) is fixed.

At this time, so that the cylinder flange 13 is not separated from the piston fixing member 80, the locking protrusion 87 protrudes from the other end of the lower plate 86 toward the upper plate 84 or the lower plate 86 at the other end of the upper plate 84. It may be formed to protrude to the side. However, in the present specification, for convenience of explanation, as shown in FIG. 10 , it will be described that the locking jaw is formed at one end of the lower plate 86 .

Accordingly, the user can open the upper plate 84 and the lower plate 86 to couple or separate the cylinder flange 13 to the flange coupling groove 89 .

Meanwhile, the piston fixing part 81 is connected to the upper plate 84 and the lower plate 86 . In more detail, the piston fixing part 81 extends from one end of the upper plate 84 and the lower plate 86 toward the front side of the cylinder body 10 and again extends to the rear of the cylinder body 10 to have a U-shape. connected by part 83 . Accordingly, the piston fixing part 81 is elastically supported by the bending part 83 .

As shown in FIG. 10 , the piston fixing part 81 is fitted into a plurality of piston fixing grooves 33 formed in the longitudinal direction of the piston body 32 . At this time, the plurality of piston fixing grooves 33 may be arranged at regular intervals.

In addition, the plurality of piston fixing grooves 33 may be formed to have different specific intervals. In particular, when a predetermined interval is required for the piston 30 to discharge the first solution 70 or the second solution 72 , a plurality of piston fixing grooves 33 may be formed accordingly. Through this, there is an advantage that the piston 30 can be moved by the same length no matter what user uses it.

The plurality of piston fixing grooves 33 cannot be moved backward by the piston fixing member 80 as shown in FIG. 10, but are formed to have a rear inclined surface so as to be movable forward.

At this time, as the piston 30 moves forward, the pressure inside the cylinder body 10 increases to generate an external force for pressing the piston 30 rearward, so that it is more stably fixed by the piston fixing member 80 .

Meanwhile, since the user is elastically supported by the bending part 83 , the piston 30 can be moved backward in a state in which the piston fixing part 81 is separated from the plurality of piston fixing grooves 33 .

At this time, guides may be provided on both sides of the piston fixing part 81 so that the piston body 32 does not shake as shown in FIG. 11 .

On the other hand, the method of extracting a nucleic acid using the nucleic acid extraction apparatus 1" according to the third embodiment of the present invention includes the process of extracting the nucleic acid using the nucleic acid extraction apparatus 1 according to the first embodiment and the second method. Since a person skilled in the art can easily understand the process of extracting nucleic acids using the nucleic acid extraction apparatus 1' according to the second embodiment, detailed descriptions will be omitted.

Although the nucleic acid extraction apparatus according to various embodiments of the present invention has been described above, the nucleic acid extraction apparatus according to this embodiment is not applied only to the field of extracting nucleic acids, and the type of filter member 20 or the first solution ( 70) or that the second solution 72 can be used as a device for extracting extracellular vesicles, specific substances, substances with different molecular weights, etc. in addition to nucleic acids in addition to nucleic acids, common knowledge in the art to which the present invention belongs Those who have it will be able to understand it clearly.

Furthermore, as the syringe device 1 from which the filter member 20 is removed, a device for stepwise dispensing of an injection solution in a necessary field, for example, in the medical field, when discharging various types of fluids step by step using a syringe, the same amount of fluid Those of ordinary skill in the art to which the present invention pertains will clearly understand that it may also be used as a device for sequentially dispensing by accommodating the first space 14 to the third space 18 .

As described above, preferred embodiments according to the present invention have been reviewed, and the fact that the present invention can be embodied in other specific forms without departing from the spirit or scope of the present invention in addition to the above-described embodiments is one of ordinary skill in the art. It is obvious to them. Therefore, the above-described embodiments are to be regarded as illustrative rather than restrictive, and accordingly, the present invention is not limited to the above description, but may be modified within the scope of the appended claims and their equivalents.

1 Syringe device 44 Central part
2 Nucleic acid extraction apparatus 46 frame
10 Cylinder body 48 Inclined surface
12 Fluid inlet 50 Cylinder case
13 Cylinder flange 52 Third thread
14 first space 54 groove
16 second space 60 cover part
18 3rd space 62 4th thread
20 filter element 70 first solution
22 filter support member 72 second solution
26 Second thread 74 Cell lysate
30 Piston 80 Piston fixing member
32 Piston body 81 Piston fixing part
34 gasket 82 guide
36 First Thread 83 Bend
38 Handle 84 Top
39 Support member 85 Opening
40 First packing member 86 Lower plate
41 Second packing member 87 Claw
42 hole 89 flange mating groove

Claims (11)

a cylinder body having a fluid outlet on the front side and a piston installed on the rear side;
a first packing member partitioning the inner space of the cylinder body into a first space adjacent to the fluid outlet and a second space adjacent to the piston rather than the first space, the first packing member being movable forward and backward in the inner space of the cylinder body;
including,
The first packing member is
a central portion formed in a disk shape having an area smaller than a cross-section perpendicular to a longitudinal direction of the inner space of the cylinder body;
a hole formed in the center of the central portion of the first packing member; and
a ring-shaped rim portion formed around the central portion of the first packing member and in close contact with the inner circumferential surface of the cylinder body; including,
The rim portion of the first packing member is moved forward in a state where the central portion of the first packing member is located behind the rim portion of the first packing member to be disposed inside the rim portion of the first packing member. 1 having an inclined surface that elastically supports the central portion of the packing member,
In the hole of the first packing member, as the piston moves forward, the fluid located in the first space is discharged to the outside through the fluid outlet, and the edge portion of the first packing member is the front end of the cylinder body. After reaching , the central portion of the first packing member is disposed inside the edge portion of the first packing member so that the fluid located in the second space is opened to be discharged to the outside through the fluid outlet through the first space Being a syringe device. The method of claim 1,
It is installed between the first packing member and the piston to partition the inner space of the cylinder body into the second space and a third space adjacent to the piston rather than the second space, to the front and rear sides in the inner space of the cylinder body a movable second packing member; further comprising,
The second packing member is
a central portion formed in a disk shape having an area smaller than a cross-section perpendicular to a longitudinal direction of the inner space of the cylinder body;
a hole formed in the center of the central portion of the second packing member; and
a ring-shaped rim portion formed around the central portion of the second packing member and in close contact with the inner circumferential surface of the cylinder body; including,
The rim portion of the second packing member is moved forward in a state where the central portion of the second packing member is located behind the rim portion of the second packing member to be disposed inside the rim portion of the second packing member. 2 and an inclined surface that elastically supports the central portion of the packing member,
In the hole of the second packing member, as the piston moves forward, the fluid located in the second space is discharged to the outside through the fluid outlet, and the edge of the second packing member is the edge of the first packing member After reaching the portion, the central portion of the second packing member is disposed inside the edge portion of the second packing member, so that the fluid located in the third space passes through the second space and the first space to the fluid outlet. A syringe device, which is open to be discharged to the outside through. delete The method of claim 1,
the piston is
a piston body fitted in the inner space of the cylinder body and movable forward or backward;
Doedoe in close contact with the inside of the cylinder body, including a gasket coupled to the front end of the piston body, the syringe device. 5. The method of claim 4,
The cylinder body is
and a first screw thread formed toward the inner side of the cylinder body at the rear portion of the cylinder body,
the piston is
a second screw thread formed on the piston body to be screw-coupled to the first screw thread; and
a handle formed at the other end of the piston body;
A syringe device comprising a. 5. The method of claim 4,
The cylinder body is coupled therein, the rear surface is opened, a hole is formed in the front surface so that the front end of the cylinder body protrudes to the outside in a state in which the cylinder body is coupled, the cylinder having a third screw thread on the outside case;
including,
a cover part having a fourth screw thread formed therein so as to be inserted into the rear part of the cylinder case so as to be screw-coupled to the third screw thread;
including,
the piston is
a support member supported inside the cover part and formed at the other end of the piston body so that the piston is pressed forward according to the rotation of the cover part;
A syringe device comprising a. 7. The method of claim 6,
The cylinder case
A syringe device in which a groove is formed in the longitudinal direction so as to confirm a position in which the gasket moves in the cylinder body. 5. The method of claim 4,
One end of the upper plate and the lower plate is connected while maintaining a predetermined distance so that a flange coupling groove is formed therebetween, and an opening is formed in the upper plate and the lower plate so that the cylinder body is fitted to the other end piston fixing member;
including,
The cylinder body is
and a cylinder flange extending outwardly around the rear end of the cylinder body so as to be fitted into the flange coupling groove,
the piston body
and a plurality of piston fixing grooves formed in the piston body at predetermined intervals in the longitudinal direction,
The piston fixing member is
a piston fixing part coupled to the plurality of piston fixing grooves to support the piston body;
a bending part connecting the piston fixing part to one end of the upper plate and the lower plate but elastically supporting the piston fixing part;
comprising, a syringe device. 3. The method according to claim 1 or 2,
a filter member connectable to the cylinder body to filter the fluid passing through the fluid outlet;
including,
wherein the filter member is coupled to a filter support member that can be coupled to the front of the cylinder body such that the filter member is disposed on the front face of the fluid outlet. A nucleic acid extraction device using the syringe device according to claim 2, comprising:
a filter member formed to filter the cell lysate containing the nucleic acid accommodated in the first space through the fluid outlet to leave the nucleic acid and cellular foreign matter in the first space;
and a filter support member coupled to the front of the cylinder body while being coupled to the filter member so that the filter member is disposed on the front surface of the fluid outlet;
The first solution as a wash buffer is accommodated in the second space,
In the third space, a second solution as an elution buffer is accommodated, a nucleic acid extraction device. As a nucleic acid extraction method using the nucleic acid extraction apparatus according to claim 10,
a cell lysate inflow step of separating the filter member and reversing the piston to introduce the cell lysate into the first space through the fluid outlet;
a cell lysate filtering step of coupling the filter member to the fluid outlet, and discharging the cell lysate accommodated in the first space by charging the piston;
a first solution discharging step of advancing the piston to discharge the first solution accommodated in the second space through the first space through the fluid outlet;
a second solution discharging step of advancing the piston to discharge the second solution accommodated in the third space into the first space through the second space; and
a nucleic acid solution discharging step of advancing the piston to discharge the nucleic acid solution accommodated in the first space through the fluid outlet;
A nucleic acid extraction method comprising a.

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