KR20230087284A - Nucleic acid xtraction pre-processing automation system - Google Patents

Abstract

An automated system for nucleic acid extraction pretreatment is disclosed. The nucleic acid extraction preprocessing automation system is a preprocessing automation system for extracting purified nucleic acids (DNA, RNA) from a specimen, wherein a syringe column into which the specimen is injected is installed while silica beads for adsorbing nucleic acids are accommodated therein, , A lysis buffer for dissolving the sample in the syringe column is provided, a wash buffer for washing the sample, an elution buffer for purifying the sample, and finally an extraction tube for accommodating the purified sample is provided. A cartridge module including a cartridge to be installed and a cartridge transfer unit for horizontally moving the cartridge on a rail; In a state of being coupled with the syringe column disposed on the cartridge, the lysis buffer and sample are pumped and injected into the syringe column, or the lysis buffer and sample accommodated in the syringe column are discharged, and the wash buffer and elution into the syringe column a syringe pump module including a syringe pump providing a suction force for sucking the buffer, and a syringe pump transfer unit disposed on one side of the syringe pump to vertically move the syringe pump; A fixing frame to which the cartridge module and the syringe pump module are fixed; may include.

Description

Nucleic acid extraction preprocessing automation system {NUCLEIC ACID XTRACTION PRE-PROCESSING AUTOMATION SYSTEM}

The present invention relates to an automated preprocessing system for nucleic acid extraction, and more specifically, as an automated preprocessing system for extracting purified nucleic acids (DNA, RNA) from a specimen, wherein the occurrence of sample contamination is prevented in the process of extracting nucleic acid from a specimen. The present invention relates to an automated system for preprocessing of nucleic acid extraction, in which the nucleic acid extraction process can be performed quickly and accurately, and the nucleic acid extraction process can be performed automatically while ensuring the safety of the inspector.

Recently, in the field of disease diagnosis, treatment strategy establishment, treatment monitoring, etc., the importance of research on the analysis and interpretation of genetic information such as genome or DNA, transcriptome, RNA, etc. is increasing.

Isolation of nucleic acids (DNA and RNA) from biological samples is the most important step in biochemical research and diagnostic processes. This is because the next steps, such as gene detection, gene cloning, gene sequencing, gene amplification, and cDNA synthesis, cannot be performed unless the nucleic acid, which is a genetic material, is isolated from the sample. Effective and reproducible isolation methods are required to differentiate DNA or RNA from different cell mixtures.

As a conventional method for extracting nucleic acids, a method of separating nucleic acids adsorbed by a magnet using magnetic beads, a method of eluting a solution using a column, a method of pushing the solution by applying air to the column, and a method of centrifuging the column itself There was a method of separating and eluting the solution.

However, nucleic acid extraction equipment using the above method has a relatively large size to process many samples, but there is a problem in that the sample processing period is excessively long. In addition, contamination due to each sample may occur in the process of processing many samples, which reduces processing efficiency and causes inconvenience to users.

The most commonly used nucleic acid extraction method is a nucleic acid extraction method using the principle of adsorption chromatography. This enables adsorption by placing a silica membrane on the extraction kit or by coating the magnetic beads with silica. In an extraction solution environment with low pH, high ionic strength, and chaotropic salts, nucleic acids of negative (-) polarity are adsorbed on the silica surface. buffer) is a method of elution.

However, when these chaotropic salts are included in the nucleic acid residue product, they act as reaction inhibitors in the PCR reaction in which the nucleic acid is finally used, so it has to go through several washing steps in the purification process, which makes the process cumbersome.

On the other hand, in the past, a 'syringe-type nucleic acid extraction device, nucleic acid extraction kit, and nucleic acid extraction method using the same (Korean Patent Publication No. 10-2021-0144999)', which can purify nucleic acids simply and quickly without the help of a separate instrument, has been proposed. has been

However, these conventional 'syringe-type nucleic acid extraction devices, nucleic acid extraction kits, and nucleic acid extraction methods using the same'

In order to extract nucleic acids using a syringe-type nucleic acid extraction device,

(a) preparing a sample by dissolving it in a solution

(b) inserting the tip of the syringe-type nucleic acid extraction device into the sample prepared in step (a), and adsorbing the nucleic acid in the prepared sample to the nucleic acid purification filter by reciprocating the piston of the syringe unit up and down

(c) washing the nucleic acid purification filter by injecting the tip part into the washing liquid and reciprocating the piston of the syringe part up and down.

(d) drying the nucleic acid purification filter by reciprocating the piston of the syringe unit up and down (e) injecting the tip unit into the nucleic acid eluate and separating the nucleic acid in the nucleic acid purification filter by reciprocating the piston of the syringe unit up and down contains

As such, in the conventional syringe-type nucleic acid extraction device and method, in carrying out the steps of dissolving the sample and isolating the nucleic acid, the tester directly intervenes to perform the overall process, such as manually preparing the sample and introducing it into the sample. There is a problem that must be done. That is, there is a problem in that it is vulnerable to sample contamination and the test accuracy is low.

Republic of Korea Patent Publication 10-2021-0144999

An object of the present invention is to solve this problem, as an automated preprocessing system for extracting purified nucleic acids (DNA, RNA) from a specimen, which minimizes the intervention of a tester in the process of extracting nucleic acids from a specimen, resulting in sample contamination. Provides an automated system for nucleic acid extraction preprocessing that prevents nucleic acid extraction in advance, enables rapid and accurate nucleic acid extraction, and prevents contact between the tester and the sample so that the nucleic acid extraction process can be performed automatically while ensuring the safety of the tester. is to do

An automated preprocessing system for nucleic acid extraction according to an embodiment of the technical idea of the present invention for achieving the above object is an automated preprocessing system for extracting purified nucleic acids (DNA, RNA) from a specimen by adsorbing the nucleic acid therein. A syringe column into which samples are injected is installed in a state where silica beads for sample are accommodated, a lysis buffer for dissolving the sample in the syringe column, a wash buffer for washing the sample, and an elution buffer for purifying the sample. a cartridge module including a cartridge in which an extraction tube for accommodating a finally purified specimen is installed, and a cartridge transfer unit for horizontally moving the cartridge on a rail; In a state of being coupled with the syringe column disposed on the cartridge, the lysis buffer and sample are pumped and injected into the syringe column, or the lysis buffer and sample accommodated in the syringe column are discharged, and the wash buffer and elution into the syringe column a syringe pump module including a syringe pump providing a suction force for sucking the buffer, and a syringe pump transfer unit disposed on one side of the syringe pump to vertically move the syringe pump; A fixing frame to which the cartridge module and the syringe pump module are fixed; may include.

In addition, the cartridge is made in the form of a rectangular block with a hollow inside, and the syringe column is connected to the column standby part for preparing the syringe column in a state where the lower portion of the syringe column is inserted on the upper surface, and the syringe pump. A column coupling part for combining columns, a lysis buffer chamber accommodating lysis buffer, a groove-type wash buffer chamber accommodating wash buffer, a groove-type elution buffer chamber accommodating elution buffer, and accommodating purified samples. An extraction tube mounting portion in which an extraction tube to be mounted may be provided.

In addition, the cartridge further includes a connection tube having one side connected to a lower end of the column coupling part and the other side connected to a lower end of the lysis buffer chamber, and when the syringe column is coupled to the column coupling unit, the connection tube Through this, the lysis buffer chamber and the syringe column are in a state of communication with each other, and the lysis buffer chamber is fitted into the open top of the lysis buffer chamber to seal the lysis buffer chamber to maintain a vacuum state, and the lysis buffer chamber It may further include a chamber push bar for pressurizing the fluid contained in the syringe column by receiving pressure or sucking the fluid contained in the syringe column into the lysis buffer chamber.

In addition, the cartridge may further include a vortex generator installed at a lower end of the lysis buffer chamber to generate a vortex in the fluid accommodated in the lysis buffer chamber.

In addition, the vortex generator includes a rotational motor generating rotational force; and a vortex generating member coupled to the rotary shaft of the rotary motor to rotate and disposed in the inner space of the lysis buffer chamber to generate a vortex in the fluid accommodated in the lysis buffer chamber.

In addition, the column coupling part may have a groove formed corresponding to the shape of the lower portion of the syringe column, and may further include a closing valve at the lower portion to prevent fluid from flowing in from the lysis buffer chamber.

In addition, the sealing valve is disposed at the lower end, a spring is inserted into the inner space, and a compression space portion into which a 'spherical'-shaped seal is inserted at the upper end of the spring; a spacer coupled to the upper portion of the compression space and penetrating the center; A slide space portion having a structure in close contact with the upper portion of the spacer member and integrated with the lower portion of the column coupling portion and having a space therein, through which the center of the upper surface passes; And a vertical cross section that is accommodated in the slide space and slides in a vertical direction, pressurizes the sealing hole as the syringe column is coupled to allow the fluid contained in the lysis buffer chamber to flow in, and has a 'T' shape. A pressure pin; may be included.

In addition, the cartridge transfer unit, generating a rotational force and a horizontal transfer motor equipped with a gear on the rotation shaft; a rack member having one side coupled to the cartridge, having a toothed rack formed on one side thereof, and engaging the rack member with a gear of the horizontal transfer motor; and a rail disposed under the fixing frame and guiding the cartridge to move horizontally along a predetermined direction in a seated state.

In addition, the syringe pump, the housing is provided with a space inside and forming an external shape; a drive motor disposed on an upper side of the housing, providing a rotational force, and having a drive pulley on a rotation shaft; A subordinate pulley disposed on a lower side spaced apart from the driving pulley; a belt wound around the drive pulley and the slave pulley; a column coupling member fixed to one side of the belt and having an end coupled to an upper portion of the syringe column; and a pump push bar fixed to the other side of the belt and compressing the lysis buffer chamber.

In addition, the column coupling member is formed in a cylindrical shape with a hollow inside, and an end portion of the cylinder is formed with a coupling groove for coupling the upper portion of the syringe column; and a piston inserted into the cylinder.

In addition, the pump push bar further includes a push bar coupling member installed at an end of the bar-shaped push rod, and the push bar coupling member has a vertical through hole formed therein, and a spring having elasticity in the through hole. It is built in, and the push rod can be inserted and coupled to slide on the push bar coupling member.

In addition, the fixing frame, the bottom portion to which the cartridge and the cartridge transfer unit is installed; 'B'-shaped side portions disposed on both sides of the cartridge module and the syringe pump module and erected vertically; and a connecting portion disposed between the bottom portion and the side portion.

The automated system for preprocessing of nucleic acid extraction according to the present invention minimizes the intervention of the inspector in the process of extracting nucleic acid from a specimen to prevent specimen contamination in advance, and simplifies the process of extracting nucleic acid from a specimen to quickly and accurately extract nucleic acids. This can be achieved, and the nucleic acid extraction process can be performed automatically in a state in which the tester's safety is secured by blocking contact between the tester and the sample.

1 is a perspective view of an automated system for nucleic acid extraction pretreatment according to an embodiment of the present invention.
2 to 10 are diagrams showing the operating state of the automated nucleic acid extraction pretreatment system according to an embodiment of the present invention.
11 and 12 are enlarged views of the main part of the closing valve showing the state before and after the syringe column is coupled to the column coupling part.

In order to fully understand the present invention and its operational advantages and objectives achieved by the practice of the present invention, reference should be made to the accompanying drawings illustrating preferred embodiments of the present invention and the description in the drawings.

Hereinafter, the present invention will be described in detail by describing preferred embodiments of the present invention with reference to the accompanying drawings. Like reference numerals in each figure indicate like elements.

An automated nucleic acid extraction preprocessing system according to an embodiment of the technical idea of the present invention is an automated preprocessing system for extracting purified nucleic acids (DNA, RNA) from a specimen, and minimizes the intervention of a tester in the process of extracting nucleic acids from a specimen. Preprocessing automation of nucleic acid extraction to prevent sample contamination in advance, ensure rapid and accurate nucleic acid extraction, and ensure safety of the tester by blocking contact between the tester and the sample so that the nucleic acid extraction process can be performed automatically It's about the system.

1 is a perspective view of an automated system for preprocessing of nucleic acid extraction according to an embodiment of the present invention, and FIGS. 2 to 10 are diagrams showing an operating state of the automated system for preprocessing of nucleic acid extraction according to an embodiment of the present invention, and FIGS. 12 is an enlarged view of the main part of the closing valve showing the state before and after the syringe column is coupled to the column coupling part.

1 to 12, the automated system for nucleic acid extraction preprocessing according to an embodiment of the present invention may largely include a cartridge module 10, a syringe pump module 20, and a fixing frame 30.

First, in the cartridge module 10, a syringe column (S) into which a sample is injected may be installed in a state where silica beads (not shown) for adsorbing nucleic acids (DNA, RNA) are accommodated therein.

The syringe column (S) may be formed of a cylindrical body (1) with a hollow inside, an upper end is open, and a nozzle (2) is formed with a narrower diameter at the lower end. In the syringe column (S), samples are delivered and accommodated, and nucleic acids can be extracted by adsorbing silica beads (not shown) provided therein.

Here, the silica beads are fine particle beads (small beads), and are used to adsorb nucleic acids instead of extracting nucleic acids using a conventional membrane-type filter, and a predetermined amount is accommodated in the syringe column (S). It plays a role in adsorbing nucleic acids.

On the other hand, the cartridge module 10 is provided with a lysis buffer (dissolution buffer) for dissolving the specimen in the syringe column (S) and a wash buffer (wash buffer) for washing the specimen, and for purifying the specimen. A cartridge 100 equipped with an elution buffer (elution buffer) for final purification and an extraction tube (T) for accommodating the finally purified sample, and a cartridge transfer unit for horizontally moving the cartridge 100 on a rail 230 ( 200) may be included.

That is, the cartridge module 10 can be provided with each buffer required for the dissolution-washing-elution process of extracting nucleic acids from a specimen.

The cartridge 100 is made in the form of a square block with a hollow inside, a column standby unit 110 for preparing the syringe column (S) with the lower portion of the syringe column (S) inserted into the upper surface, A column coupling part 120 for coupling the syringe column S while being coupled with the syringe pump S, a lysis buffer chamber 140 for accommodating a lysis buffer, and a groove for accommodating a wash buffer. A wash buffer chamber 160, an illusion buffer chamber 170 in the form of a groove in which an illusion buffer is accommodated, and an extraction tube holder 180 in which an extraction tube T accommodating a purified specimen are mounted may be provided.

That is, on the upper surface of the cartridge 100, the column standby unit 110, the column coupling unit 120, the lysis buffer chamber 140, the wash buffer chamber 160, the illusion buffer chamber 170 and the extraction tube holder ( 180) may be provided to be positioned adjacently.

The column waiting part 110 is provided on one side of the upper surface of the cartridge 100, and the lower part of the syringe column S can be inserted and fixed, that is, the nozzle 2 The groove is formed to correspond to the shape of the syringe column (S) so that the portion including the portion and the body portion (1) can be inserted and fixed.

The column standby unit 110 is installed by an operator to be prepared in a standby state before combining with the syringe pump module 20 to be described later, and the syringe column (S) coupled to the column standby unit 110 As the syringe pump module 20 operates, the silver may be drawn out from the column waiting unit 110 and moved.

The column coupling part 120 is a part where the syringe column S coupled to the column waiting part 110 is moved by the syringe pump module 20 and coupled to inject a sample.

The column coupling part 120 may be formed in the form of a groove similar to the shape of the column waiting part 110, but the end of the groove may be formed in an open form.

The lysis buffer chamber 140 is installed at a position adjacent to the column coupling part 120, and is a part where the lysis buffer and the sample are accommodated.

That is, the lysis buffer chamber 140 is a part where a liquid or particulate lysis buffer and a sample are accommodated, and the received lysis buffer and a sample are directed into the syringe column (S) coupled to the column coupling part 120. It is passed on.

To this end, the lysis buffer chamber 140 further includes a connection pipe 141 having one side connected to the lower end of the column coupling part 120 and the other side connected to the lower end of the lysis buffer chamber 140, , When the syringe column (S) is coupled to the column coupling part 120, the lysis buffer chamber 140 and the syringe column (S) can be in a state of mutual communication through the connection pipe 141. .

Meanwhile, the lysis buffer chamber 140 is inserted into the open upper portion of the lysis buffer chamber 140 to seal the lysis buffer chamber 140 to maintain a vacuum state, and the lysis buffer chamber 140 A chamber push for pumping fluids (lysis buffer and sample) contained in ) to the syringe column (S) under pressure or sucking the fluid contained in the syringe column (S) into the lysis buffer chamber (140). A bar 142 may be further included.

The chamber push bar 142 seals the open top of the lysis buffer chamber 140 and serves as a kind of piston for pressurizing the fluid contained in the lysis buffer chamber 140 to the syringe column (S) As such, it may be formed in a rod shape that is closely fitted to the inner circumferential surface of the lysis buffer chamber 140 and protrudes upward.

At this time, as shown in FIGS. 11 and 12, the column coupling part 120 has a groove formed corresponding to the lower shape of the syringe column S, and the syringe column S ) may further include a sealing valve 130 for regulating fluid flow from the lysis buffer chamber 140 in a state in which it is not coupled.

The sealing valve 130 is disposed at the lower end of the groove of the column coupling part 120, a spring 131 is inserted into the inner space, and a 'spherical' shaped closing hole 132 is inserted at the upper end of the spring 131. A compression space part 133, a spacer 134 closely coupled to the upper part of the compression space part 133 and having a through hole 135 through the center thereof, and a spacer 134 in close contact with the upper part of the spacer 134, but the column It is made of a structure integrated with the lower part of the coupling part 120, and a space is provided inside, and the slide space part 136 through which the center of the upper surface penetrates and is accommodated in the slide space part 136 and slides in the vertical direction. And, as the syringe column (S) is coupled, the sealing hole 132 is pressurized to open the fluid accommodated in the lysis buffer chamber 140 to flow in. A pressing pin 137 having a 'T' shape ) may be included.

That is, the sealing valve 130 is a fastening means (eg, a bolt, not shown) in a state in which the compression space portion 133, the spacer member 134, the slide space portion 136, and the pressure pin 137 are in close contact with each other. It can be coupled to the lower end of the column coupling part 120 by.

The compression space 133 accommodates the spring 131 and the seal 132 in the through-hole, and one side of the connection pipe 141 can be fitted to an end having a smaller diameter than the central diameter. . The compression space portion 133 has a flange shape having a larger diameter than the center portion, and a hole for inserting a fastening means such as a bolt may be formed in the flange portion.

The spacer 134 has a circular plate structure, and a through hole 135 is formed in the center portion, and the press pin 137 is inserted into the upper and lower sides of the through hole 135 to provide airtightness when sliding. A sealing member (unsigned) may be installed to maintain. In addition, a hole for inserting a fastening means such as a bolt may be formed in the rim portion.

The slide space part 136 is in close contact with the upper part of the spacer member 134 but has a structure integrated with the lower part of the column coupling part 120, and has an internal space, but the upper part and the lower part are opened respectively. , and the diameter of the open hole is smaller in the upper part than in the lower part, so that the pressing pin 137, which will be described later, slides vertically in the inner space of the slide space part 136, the upper part of the slide space part 136. You can get caught in it and prevent it from escaping. Accordingly, the upper hole diameter of the slide space portion 136 may be formed smaller than the upper diameter of the pressing pin 137 . In addition, a hole for inserting a fastening means such as a bolt may be formed in the rim portion.

The pressure pin 137 allows the fluid contained in the lysis buffer chamber 140 to flow into the syringe column S in a state in which the syringe column S is coupled to the column coupling part 120. As a part for opening the passage, a flange 138 having a larger upper diameter than the lower portion is provided, and the lower flange 138 has a relatively small diameter and is inserted into the through hole 135 of the spacer 134. In this state, a sliding motion may be performed vertically within the slide space 136 . That is, when the nozzle 2 of the syringe column S presses the upper surface of the pressure pin 137, the pressure pin 137 moves downward, and accordingly, the lower end of the pressure pin 137 moves upward. When the sealing port 132 is pushed downward, the space between the inner space accommodating the sealing port 132 and the sealing port 132 is opened, and when suction force on the syringe column (S) side is generated, the lysis buffer chamber 140 The fluid contained in can be transferred to the syringe column (S).

At this time, an O-ring 139 is provided at the lower end of the pressure pin 137 at a portion in contact with the sealing hole 132 to maintain watertightness.

Meanwhile, the cartridge 100 may further include a vortex generating device 150 installed at a lower end of the lysis buffer chamber 140 and generating a vortex in the fluid accommodated in the lysis buffer chamber 140. .

The vortex generating device 150 generates vortexes in the sample space accommodated in the lysis bumper chamber 140 to facilitate separation of nucleic acid from the sample accommodated in the lysis bumper chamber 140 .

Here, the sample initially accommodated in the lysis bumper chamber 140 means everything including the sample material adsorbed to the sample object (eg, cotton swab, cotton, etc.) for collecting the first sample material, lysis bumper After inserting the sample material adsorbed into the lysis bumper chamber 140 containing the sample material, the sample material can be easily separated from the sample material through the vortex generator 150.

The vortex generator 150 is coupled to a rotational motor 151 generating rotational force and a rotational shaft of the rotational motor 151 to rotate, and is disposed in the inner space of the lysis buffer chamber 140 to form the lysis buffer chamber It may include a vortex generating member 152 that generates a vortex in the fluid accommodated in 140.

The rotary motor 151 may be disposed below the lysis bumper chamber 140 , and the vortex generating member 152 may be installed on a rotation shaft of the rotary motor 151 .

At this time, the vortex generating member 152, although not specifically shown in the drawing, rotates according to the rotational force of the rotation motor 151, and the lower bottom surface of the lysis bumper chamber 140 is separated from the outer circumferential portion and is rotatable It may be a structural configuration, and at this time, a vane may protrude on the bottom surface to form a vortex of fluid.

In addition, a separate cylindrical component may be inserted into the bottom surface and the inner circumferential surface of the lysis bumper chamber 140 to rotate together with the rotation of the rotary motor 151 to generate eddies.

That is, the vortex generating device 150 may be applied in various forms as long as it has a configuration capable of generating vortex in the inner space of the lysis bumper chamber 140 by using the rotational force of the rotation motor 151 .

Meanwhile, the wash buffer chamber 160 is formed on the upper surface of the cartridge 100 adjacent to the column standby part 110, and has a volume corresponding to the volume of the wash buffer to be injected into the syringe column S. It can be formed in the shape of a groove.

Meanwhile, the illusion buffer chamber 170 is formed on the upper surface of the cartridge 100 adjacent to the wash buffer chamber 160, and has a volume corresponding to the volume of the illusion buffer to be injected into the syringe column S. shape can be formed.

On the other hand, the extraction tube holder 180 is formed on the upper surface of the cartridge 100 adjacent to the illusion buffer chamber 170, and the extraction tube (T) accommodates the purified sample stored in the syringe column (S). ) is the part to be mounted. That is, the extraction tube T, which is a container for receiving the finally extracted nucleic acid and transferring it to a separate testing facility (not shown), is prepared.

On the other hand, the cartridge transfer unit 200 generates a rotational force and a horizontal transfer motor 210 in the form of a stepping motor equipped with a gear on a rotating shaft, one side is coupled to the cartridge 100, a sawtooth rack is formed on one side, , The rack is disposed under the rack member 220 coupled to engage with the gear of the horizontal transfer motor 210 and the fixed frame 30 to be described later, and moves horizontally along a predetermined direction while the cartridge 100 is seated. It may include a rail 230 for guiding to do so.

That is, when the horizontal transfer motor 210 rotates, the rack member 220 moves horizontally in one direction, and accordingly, the cartridge 100 can move horizontally together. Thus, the cartridge 100 can be moved to a set position through the cartridge transfer unit 200 .

Next, the syringe pump module 20 may largely include a syringe pump 300 and a syringe pump transfer unit 300 .

The syringe pump 300 has a housing 310 having a space inside and forming an external shape, a driving motor disposed on an upper inside of the housing 310 to provide a rotational force, and a driving pulley 321 provided on a rotational shaft. 320, a slave pulley 330 disposed on a lower side spaced apart from the drive pulley 321, a belt 340 wound around the drive pulley 321 and the slave pulley 330, one side of the belt 340 and a column coupling member 350 having an end coupled to an upper portion of the syringe column S and a pump push bar 360 fixed to the other side of the belt 340 and compressing the lysis buffer chamber 140. can do.

That is, the syringe pump 300 pressurizes the fluid in the lysis buffer chamber 140 to the syringe column (S) by using the rotational force of the driving motor 320 or, conversely, to the syringe column (S). It serves to pressurize the received fluid into the lysis buffer chamber 140, and when the drive motor 320 rotates in one direction, the pump push bar 360 moves the chamber of the lysis buffer chamber 140. The fluid can be pressurized by pressing the push bar 142, and when the drive motor 320 reversely rotates in the other direction, the column coupling member 350 transfers the fluid in the syringe column S to the lysis buffer chamber. It becomes possible to pressurize to the (140) side.

Here, the column coupling member 350 is formed in a cylindrical shape with a hollow inside, and a cylinder 351 having a coupling groove formed at an end for coupling the upper portion of the syringe column (S) and inserted into the cylinder 351 A piston 352 may be included.

That is, as the drive motor 320 rotates, the belt 340 moves and the piston 352 moves up and down, so that the fluid in the syringe column S can be pumped or sucked.

In addition, the pump push bar 360 further includes a push bar coupling member 362 installed at an end of the bar-shaped push rod 361, and the push bar coupling member 362 has a vertical through hole formed therein. And, a spring having elasticity is built into the through hole, and the push rod 361 is inserted on the push bar coupling member 362 and coupled to slide.

On the other hand, the syringe pump transfer unit 400 is a part for vertically transferring the syringe pump 300, and receives power from a power source such as a motor while being coupled to one side of the syringe pump 300 to move up and down. Part. Although not specifically shown in the drawings, various shapes may be applied as long as they are configured to move up and down by driving a normal motor, and the shapes are not limited.

Finally, the fixing frame 30 is a part to which the cartridge module 10 and the syringe pump module 20 are fixed.

The fixing frame 30 is disposed on both sides of the bottom part 31 where the cartridge 100 and the cartridge transfer unit 200 are installed, the cartridge module 10 and the syringe pump module 20, and is erected vertically. A 'B' shaped side part 32 and a connection part 33 disposed between the bottom part 31 and the side part 32 may be included.

That is, the fixing frame 30 may serve as a cover for fixing the cartridge module 10 and the syringe pump module 20 and protecting them from the outside, and the side part 32 may have a plurality of Holes can be drilled.

Hereinafter, referring to FIGS. 2 to 10, the operating state of the automated nucleic acid extraction pretreatment system according to the present invention will be described.

First, the syringe column S is inserted into the column standby part 110 of the cartridge 100 in a state where the cartridge 100 and the syringe pump 300 are initially separated, and the lysis buffer chamber 140 contains the lysis buffer chamber 140. In the state where the buffer and the specimen are accommodated, the wash buffer is injected into the wash buffer chamber 160, the illusion buffer is injected into the illusion buffer chamber 170, and the extraction tube T is inserted into the extraction tube holder 180. It is set up and ready to go.

Then, the cartridge 100 is transported to the position of the syringe pump 300 and coupled with the syringe column S inserted into the column waiting unit 110, and the syringe column S moves as the syringe pump transfer unit 400 operates. In this coupled state, the syringe pump 300 moves upward.

Then, when the cartridge 100 is horizontally retracted and transported to the position of the column coupling part 120, the syringe pump 300 vertically moves downward through the syringe pump transfer part 400 so that the syringe column S becomes a column. It is inserted into and coupled to the coupling part 120 .

Then, when the driving motor 320 is operated and the pump push bar 360 moves downward, the chamber push bar 142 of the lysis buffer chamber 140 moves downward, and at the same time the column coupling member 350 As the piston 352 of the rises, the fluid accommodated in the lysis buffer chamber 140 is pressurized into the syringe column (S), and a suction process is performed.

Then, when the driving motor 320 operates in the reverse direction, the piston 352 of the column coupling member 350 descends, and at the same time, when the pump push bar 360 moves upward, the fluid accommodated in the syringe column S It is pressurized into the lysis buffer chamber 140 and discharged.

In the process of inhaling and discharging the sample and the lysis buffer, the sample introduced into the syringe column (S) is sucked in and discharged together with the lysis buffer in a vacuum state, thereby dissolving the sample.

Then, the syringe pump transfer unit 400 operates to transfer the syringe pump 300 upward with the series column S coupled thereto, and then the cartridge transfer unit 200 operates so that the cartridge 100 becomes a wash buffer. It is transported to the position of the accommodated wash buffer chamber 160, and the syringe pump transfer unit 400 operates again to transport the syringe pump 300 downward.

Then, while the syringe column (S) is located in the wash buffer chamber 160, the driving motor 320 operates to suck the wash buffer into the syringe column (S).

Then, the syringe pump 300 and the cartridge 100 move vertically and horizontally through the syringe pump transfer unit 400 and the cartridge transfer unit 200 so that the syringe column S moves to the column coupling unit 120 and is coupled. After reaching the state, the wash buffer in the syringe column (S) is discharged to the lysis buffer chamber (140) by the operation of the syringe pump (300).

In this way, the washing process is performed through the suction and discharge process of the injected wash buffer in the state where the sample is accommodated in the syringe column (S) and the nucleic acid is adsorbed on the surface of the silica beads.

Then, the syringe pump 300 and the cartridge 100 move vertically and horizontally through the syringe pump transfer unit 400 and the cartridge transfer unit 200 so that the syringe column S moves to the elution buffer chamber 170 in which the elution buffer is accommodated. After moving, the driving motor 320 operates to suck the elution buffer into the syringe column (S).

Then, the syringe pump 300 and the cartridge 100 move vertically and horizontally through the syringe pump transfer unit 400 and the cartridge transfer unit 200, so that the syringe column (S) moves to the position of the extraction tube (T), and then the syringe The elution buffer containing the nucleic acid of the sample in the column (S) is discharged into the extraction tube (T).

The nucleic acid finally accommodated in the extraction tube T is transferred to a separate testing facility for additional testing.

Therefore, it is possible to quickly and accurately perform the dissolution-washing-elution process of the sample through the automated system for nucleic acid extraction pretreatment of the present invention.

The automated nucleic acid extraction preprocessing system according to various embodiments according to the technical idea of the present invention minimizes the intervention of a tester in the process of extracting nucleic acid from a sample to prevent sample contamination in advance and extracts nucleic acid from the sample. The nucleic acid extraction process can be performed quickly and accurately by simplifying the process, and the nucleic acid extraction process can be performed automatically while ensuring the safety of the inspector by blocking contact between the tester and the sample.

As described above, the optimum embodiment has been disclosed in the drawings and specifications. Although specific terms are used herein, they are only used for the purpose of describing the present invention and are not used to limit the meaning or the scope of the present invention described in the claims. Therefore, those of ordinary skill in the art will understand that various modifications and equivalent other embodiments are possible therefrom. Therefore, the true technical protection scope of the present invention should be determined by the technical spirit of the appended claims.

S: syringe column 1: body part
2: nozzle T: extraction tube
10: cartridge module 100: cartridge
110: column standby unit 120: column coupling unit
130: sealing valve 131: spring
132: sealing port 133: compression space
134: spacer member 135: through hole
136: slide space portion 137: pressure pin
138: flange 139: O-ring
140: lysis buffer chamber 141: connector
142: chamber push bar 150: vortex generator
151: rotation motor 152: vortex generating member
160: wash buffer chamber 170: illusion buffer chamber
180: extraction tube holder 200: cartridge transfer unit
210: horizontal transfer motor 220: rack member
230: rail 20: syringe pump module
300: syringe pump 310: housing
320: drive motor 321: drive pulley
330: dependent pulley 340: belt
350: column coupling member 351: cylinder
352: piston 360: pump push bar
361: push rod 362: push bar coupling member
400: syringe pump transfer unit 30: fixed frame
31: bottom part 32: side part
33: connection part

Claims (12)

In the preprocessing automation system for extracting purified nucleic acid (DNA, RNA) from a specimen,
A syringe column into which a sample is injected is installed in a state where silica beads for adsorbing nucleic acids are accommodated therein, a lysis buffer for dissolving the sample in the syringe column, and a wash buffer for washing the sample. A cartridge module including an illusion buffer for purifying the cartridge, a cartridge in which an extraction tube for accommodating the finally purified sample is installed, and a cartridge transfer unit for horizontally moving the cartridge on a rail;
In a state of being coupled with the syringe column disposed on the cartridge, the lysis buffer and sample are pumped and injected into the syringe column, or the lysis buffer and sample accommodated in the syringe column are discharged, and the wash buffer and elution into the syringe column a syringe pump module including a syringe pump providing a suction force for sucking the buffer, and a syringe pump transfer unit disposed on one side of the syringe pump to vertically move the syringe pump;
A nucleic acid extraction preprocessing automation system comprising a; fixing frame to which the cartridge module and the syringe pump module are fixed. According to claim 1,
the cartridge,
It is made in the form of a square block with a hollow inside,
A column standby part for preparing the syringe column with the lower part of the syringe column inserted into the upper surface, a column coupling part for combining the syringe column in a state coupled with the syringe pump, and a lysis buffer for receiving the lysis buffer. Nucleic acid extraction characterized in that a chamber, a wash buffer chamber in the form of a groove accommodating the wash buffer, an elution buffer chamber in the form of a groove accommodating the elution buffer, and an extraction tube holding portion in which the extraction tube accommodating the purified specimen is mounted Preprocessing automation system. According to claim 2,
the cartridge,
A connection pipe having one side connected to a lower end of the column coupling part and the other side connected to a lower end of the lysis buffer chamber, and when the syringe column is coupled to the column coupling unit, the lysis buffer through the connection tube The chamber and the syringe column are in communication with each other,
It is inserted into the open top of the lysis buffer chamber to seal the lysis buffer chamber to maintain a vacuum state, and pressurize the fluid contained in the lysis buffer chamber to the syringe column by receiving pressure, or Nucleic acid extraction preprocessing automation system, characterized in that it further comprises a chamber push bar for sucking the fluid into the lysis buffer chamber. According to claim 3,
the cartridge,
The nucleic acid extraction preprocessing automation system further comprising a vortex generator installed at the bottom of the lysis buffer chamber and generating a vortex in the fluid accommodated in the lysis buffer chamber. According to claim 4,
The vortex generator,
a rotational motor generating rotational force; and
A nucleic acid extraction preprocessing automation system comprising a; vortex generating member coupled to the rotating shaft of the rotary motor and rotating and disposed in the inner space of the lysis buffer chamber to generate vortex in the fluid accommodated in the lysis buffer chamber. According to claim 2,
The column coupling part,
A groove is formed to correspond to the lower shape of the syringe column,
The nucleic acid extraction pre-processing automation system further comprises a sealing valve at the bottom to prevent the fluid from flowing in from the lysis buffer chamber. According to claim 6,
The sealing valve is
a compression space portion disposed at a lower end, into which a spring is inserted into an inner space, and into which a 'spherical'-shaped seal is inserted at an upper end of the spring;
a spacer coupled to the upper portion of the compression space and penetrating the center;
A slide space portion having a structure in close contact with the upper portion of the spacer member and integrated with the lower portion of the column coupling portion and having a space therein, through which the center of the upper surface passes; and
It is accommodated in the slide space and slides in a vertical direction, and as the syringe column is coupled, the sealing hole is pressurized so that the fluid contained in the lysis buffer chamber is opened so that the fluid contained in the lysis buffer chamber is introduced. Nucleic acid extraction pre-processing automation system comprising a; pin. According to claim 1,
The cartridge transfer unit,
A horizontal transfer motor generating rotational force and having a gear on the rotational shaft;
a rack member having one side coupled to the cartridge, having a toothed rack formed on one side thereof, and engaging the rack member with a gear of the horizontal transfer motor; and
A nucleic acid extraction preprocessing automation system comprising a; rail disposed under the fixing frame and guiding the cartridge to move horizontally along a predetermined direction in a seated state. According to claim 2,
The syringe pump,
A housing having a space inside and forming an external shape;
a drive motor disposed on an upper side of the housing, providing a rotational force, and having a drive pulley on a rotation shaft;
A subordinate pulley disposed on a lower side spaced apart from the driving pulley;
a belt wound around the drive pulley and the slave pulley;
a column coupling member fixed to one side of the belt and having an end coupled to an upper portion of the syringe column; and
A nucleic acid extraction preprocessing automation system comprising a; pump push bar fixed to the other side of the belt and compressing the lysis buffer chamber. According to claim 9,
The column coupling member,
a cylinder having a hollow cylinder shape and having a coupling groove at an end portion for coupling an upper portion of the syringe column; and
A nucleic acid extraction pre-processing automation system comprising a; piston inserted into the cylinder. According to claim 9,
The pump push bar,
Further comprising a push bar coupling member installed at the end of the bar-shaped push rod,
The push bar coupling member,
A nucleic acid extraction preprocessing automation system, characterized in that a vertical through hole is formed therein, a spring having elasticity is built in the through hole, and the push rod is inserted and coupled to slide on the push bar coupling member. According to claim 1,
The fixed frame,
a bottom portion in which the cartridge and the cartridge transfer unit are installed;
'B'-shaped side portions disposed on both sides of the cartridge module and the syringe pump module and erected vertically; and
A nucleic acid extraction preprocessing automation system comprising a; connecting portion disposed between the bottom portion and the side portion.

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