KR20240019435A - Nucleic acid extraction and transfer apparatus - Google Patents

Abstract

The present invention relates to a nucleic acid extraction and nucleic acid delivery device, which includes a cartridge seating portion on which a plurality of extraction cartridges containing mixed samples and buffers are seated, and a cartridge seating portion provided adjacent to the cartridge seating portion to accommodate a nucleic acid amplification reagent (PCR reaction). A platform having a tube mounting portion on which a nucleic acid amplification reagent tube is mounted; a plunger that raises and lowers to selectively insert or extract one of the plurality of extraction cartridges mounted on the cartridge seating portion and the nucleic acid amplification reagent tube mounted on the tube seating portion; a plunger lifting part that has a plunger mounting part to which the plunger is attached and detached, and that raises and lowers the plunger; a magnetic rod that moves up and down to be inserted into or extracted from the plunger, and attaches a magnetic bead accommodated in the extraction cartridge to the plunger or separates it from the plunger; It includes a rod lifting part that elevates the commercial magnetic rod, and is recessed in the tip of the plunger to enable a certain amount of elution buffer containing nucleic acids to be moved from the extraction cartridge to the nucleic acid amplification reagent tube.

Description

Nucleic acid extraction and nucleic acid delivery device {NUCLEIC ACID EXTRACTION AND TRANSFER APPARATUS}

The present invention relates to a nucleic acid extraction and nucleic acid delivery device, and more specifically, to a nucleic acid extraction and nucleic acid delivery device that can separate and extract nucleic acids in one device and transfer the extracted nucleic acids to a nucleic acid amplification tube.

The procedure of extracting pure nucleic acids without impurities from various biological samples such as blood, plasma, saliva, urine, or tissue, and amplifying them using the extracted pure nucleic acids, is used in various fields such as molecular diagnosis and forensics.

In particular, with the recent outbreak of various infectious diseases such as Covid-19 and monkeypox, fast and simple molecular diagnosis has become increasingly important.

Currently, the most commonly used automatic molecular diagnostic procedure is to dissolve the sample in a lysis buffer capable of dissolving the sample, and then use a magnetic device capable of attaching nucleic acids to the well containing the dissolved sample and the lysis buffer. After inserting the bead and attaching the nucleic acid to the magnetic bead, only the magnetic bead with the nucleic acid attached is pulled out using a plunger with an opening only at the top and a magnet inserted through the top of the plunger, and the magnetic bead is placed in a washing buffer. After moving, remove any foreign substances on the magnetic beads.

Then, the magnetic beads in the washing buffer are recovered using a plunger and a magnet, the magnetic beads are moved to the elution buffer, and pure nucleic acids attached to the magnetic beads are eluted in the elution buffer to extract pure nucleic acids. It's rough.

This extraction process using magnetic beads involves extracting multiple samples using automatic extraction equipment using a disposable plastic plunger and a plastic cartridge or deep well plate pre-filled with various buffers necessary for the extraction process. Extract.

Illusion buffer containing the nucleic acid extracted through this process is injected into the nucleic acid amplification reagent tube through the upper opening of the nucleic acid amplification reagent tube (PCR agent tube) containing the amplification reagent using a single or multi-pipette. The upper opening of the amplification reagent tube is sealed with a lid or film, then transferred to a separately provided nucleic acid amplification device, and the amount of amplified nucleic acid is measured and diagnosed.

In this way, in order to amplify nucleic acids from a sample sample, after extracting nucleic acids from a separately provided nucleic acid extraction device, the operator takes out the extraction cartridge containing the illusion buffer containing the nucleic acids from the nucleic acid extraction device, and then extracts the nucleic acids contained in the extraction cartridge. After dispensing a fixed amount of the illusion buffer with a pipette, it must be manually injected one by one into the nucleic acid amplification reagent tube provided separately from the nucleic acid extraction device. Not only is the transfer process of the illusion buffer containing nucleic acids cumbersome, but the transfer of the illusion buffer containing nucleic acids is also cumbersome. There are problems with errors occurring or contamination during the dispensing process.

US registered patent US 5,200,084 (Title of invention: APPARATUS AND METHODS FOR MAGNETIC SEPARATION, Registration date: 1993.04.06.) PCT International Application Publication WO 2001/68263 A1 (Title of invention: VESSEL AND ROD, Publication date: 2001.09.20.)

The present invention is intended to solve the above problems, and prevents contamination of the nucleic acid to be amplified, safely separates, purifies and extracts the nucleic acid to amplify the nucleic acid, and performs the separation, purification and extraction process of the extract, and the nucleic acid delivery process. It can be implemented in one device and can be automated. In order to amplify the nucleic acid, the operator does not need to do the dispensing and transfer process of the nucleic acid to be amplified, preventing errors or contamination during the dispensing and transfer process of the nucleic acid. The purpose is to provide a nucleic acid extraction and nucleic acid delivery device that can perform nucleic acid extraction and nucleic acid delivery.

Another object of the present invention is to provide a nucleic acid extraction and nucleic acid amplification device that can amplify nucleic acids by using a heater module in the nucleic acid amplification reagent tube after nucleic acid extraction and nucleic acid delivery.

The purpose of the present invention is to provide a cartridge seating portion on which a plurality of extraction cartridges containing a mixture of samples and buffers are seated, and a plurality of nucleic acid amplification reagent tubes provided adjacent to the cart seating portion to accommodate a nucleic acid amplification reagent (PCR reaction). a platform having a tube seating portion on which to seat and amplify; a plunger that raises and lowers to selectively insert or extract one of the plurality of extraction cartridges mounted on the cartridge seating portion and the plurality of nucleic acid amplification reagent tubes mounted on the tube seating portion; a plunger lifting part that has a plunger mounting part to which the plunger is attached and detached, and that raises and lowers the plunger; a magnetic rod that moves up and down to be inserted into or extracted from the plunger, and attaches a magnetic bead accommodated in the extraction cartridge to the plunger or separates it from the plunger; This can be achieved by a nucleic acid extraction and nucleic acid amplification device characterized by comprising a rod lifting unit that raises and lowers the commercial magnetic rod.

Here, the plunger may include a recess groove that is recessed in the tip of the plunger and forms a predetermined amount of droplets of the buffer contained in the extraction cartridge.

The recess groove may have a semi-spherical, spherical or polygonal partial cross-sectional shape.

A heating module provided on the tube mounting portion and heating the nucleic acid amplification reagent tube mounted on the tube mounting portion may be further included to amplify nucleic acids.

The plunger lifting part includes a support bracket for the plunger that supports the plunger mounting part; a plunger screw shaft rotatably coupled to the plunger support bracket to raise and lower the plunger support bracket; It may include a drive motor for the plunger that rotates the screw shaft for the plunger.

The rod lifting unit includes a rod support bracket for supporting the magnetic rod; a rod screw shaft rotatably coupled to the rod support bracket to elevate and lower the rod support bracket; It may include a rod drive motor that rotates the rod screw shaft.

It further includes a reciprocating moving part that reciprocates the plunger lifting part and the rod lifting part in a horizontal direction with respect to the lifting direction of the plunger, wherein the reciprocating moving part includes a main support bracket supporting the plunger lifting part and the rod lifting part; a main screw shaft rotatably coupled to the main support bracket and reciprocating the main support bracket in a horizontal direction with respect to the ascending and descending direction of the plunger; It may include a main drive motor that rotates the main screw shaft.

According to the present invention, a plurality of extraction cartridges containing a mixed sample and a buffer, and a plurality of nucleic acid amplification reagent tubes containing a nucleic acid amplification reagent (PCR reaction) are seated in one device, and the nucleic acid extracted from the extraction cartridge By providing the nucleic acid directly to the nucleic acid amplification reagent tube provided within the device without moving it outside the device, contamination of the nucleic acid to be amplified can be prevented and the nucleic acid can be safely separated and extracted to amplify the nucleic acid, and the separation and extraction process of the extract , the nucleic acid delivery process can be implemented in one device and automation can be achieved. In order to amplify the nucleic acid, the operator does not need to dispense and transfer the nucleic acid to be amplified, eliminating the risk of errors in the process of dispensing and transferring the nucleic acid. It can be prevented from occurring or becoming contaminated.

In addition, by providing a recess groove at the tip of the plunger, the buffer contained in the extraction cartridge is stored in a certain amount in the form of droplets in the recess groove by surface tension without a pipette tip or other additional liquid delivery device. By condensing, the buffer can be dispensed quantitatively and nucleic acids can be precisely extracted and delivered.

1 is a configuration diagram of a nucleic acid extraction and nucleic acid amplification device according to an embodiment of the present invention;
Figure 2 is a bottom perspective view of the plunger of Figure 1;
Figure 3 is a longitudinal cross-sectional view of the plunger of Figure 2;
Figures 4 and 5 illustrate the process of extracting nucleic acids using the nucleic acid extraction and nucleic acid amplification device of Figure 1;
Figure 6 is a diagram showing the process of dispensing extracted nucleic acid using the nucleic acid extraction and nucleic acid amplification device of Figure 1 and injecting it into a nucleic acid amplification reagent tube;
Figures 7 (a) and (b) illustrate the process of attaching magnetic beads accommodated in a buffer to the surface of a plunger;
Figures 8 (a) and (b) are diagrams showing the process of separating magnetic beads attached to the surface of a plunger with a buffer.

Hereinafter, the present invention will be described in detail with reference to the accompanying drawings.

Figure 1 shows a nucleic acid extraction and nucleic acid amplification device according to an embodiment of the present invention.

As shown in these figures, the nucleic acid extraction and nucleic acid amplification device 1 according to an embodiment of the present invention includes a platform 30, a plunger 50, a plunger lifting part, a magnetic rod 70, Includes a load lift unit.

The platform 30 has a flat shape and includes a cartridge seating portion 31, a tube seating portion 35, and a tube heating module 36.

The cartridge seating portion 31 is provided on the upper surface of the platform 30, so that the sample and buffers 15a, 15b, 15d, lysis buffer, washing buffer, and elution buffer, and magnetic beads can be mixed and accommodated in a plurality of chambers. The extraction cartridge 10 consisting of (10a, 10b, 10c, 10d) is seated. The extraction cartridge 10 having a plurality of chambers 10a, 10b, 10c, and 10d may be fitted into the cartridge seating portion 31.

The chambers 10a, 10b, 10c, and 10d of the extraction cartridge 10 have a hollow circular cross-section tube shape with one side open. In addition, a chamber accommodating a sample or buffer (15a, 15b, 15d) and a magnetic bead (15c) is formed inside the extraction cartridge 10, which is composed of a plurality of chambers (10a, 10b, 10c, and 10d). Here, samples collectively refer to those made up of liquids, solids, or mixtures of liquids and solids in biological or chemical processes. The buffers 15a, 15b, and 15d collectively refer to a lysis buffer, a washing buffer, an elusion buffer, etc. In the drawing, reference numeral 15c refers to a solution containing magnetic beads. The sample is introduced into the chamber 10a containing the lysis buffer 15a and mixed.

The tube seating portion 35 is provided adjacent to the cartridge seating portion 31 on the upper surface of the platform 30, on which the nucleic acid amplification reagent tube 20, which accommodates the nucleic acid amplification reagent 25 (PCR reaction), is seated, and the heating module The nucleic acid in the nucleic acid amplification reagent tube (20) is amplified by (36). The nucleic acid amplification reagent tube 20 may be fitted into the cartridge seating portion 31.

Here, although not shown, the platform 30 horizontally horizontally extracts the extraction cartridge 10, which consists of a plurality of chambers 10a, 10b, 10c, and 10d provided in the cartridge seating portion 31 of the platform 30 during the nucleic acid extraction process. It can be connected to a shaking motor (not shown) that shakes by moving the trajectory. As the shaking motor drives, the sample, buffers 15a, 15b, 15d, and magnetic beads accommodated in the extraction cartridge 10, which consists of a plurality of chambers 10a, 10b, 10c, and 10d mounted on the cartridge seating portion 31. The solution 15c containing flows along the wall of the extraction cartridge 10, allowing the sample, buffers 15a, 15b, 15d, and the solution 15c to be mixed and flowed at high speed. In addition, it is possible to separate and purify high-concentration extracts from large or small amounts of samples contained in containers with large or small diameters.

As shown in FIGS. 2 and 3, the plunger 50 has a hollow circular cross-sectional tube shape with one side open. The plunger 50 has a relatively smaller diameter than the diameters of the extraction cartridge 10 and the nucleic acid amplification reagent tube 20. As a result, the plunger 50 can be inserted into the chambers 10a, 10b, 10c, and 10d of the extraction cartridge 10 and the nucleic acid amplification reagent tube 20, and the chambers 10a, 10b, and 10c, 10d) and each inner periphery of the nucleic acid amplification reagent tube 20, a space through which the sample, buffers 15a, 15b, 15d, and solution 15c can flow is formed. Additionally, a magnetic rod 70 is inserted and extracted inside the plunger 50.

Meanwhile, a recess groove 55 is formed recessed at the tip of the plunger 50, for example, at the lower end of the plunger 50.

In this embodiment, the recess groove 55 is shown as having a semi-spherical cross-sectional shape, but the present invention is not limited thereto, and the recess groove 55 may have a partial spherical or polygonal cross-sectional shape. The diameter of the lower end of the plunger 50 is smaller than the diameter of the nucleic acid amplification tube 20, so that the liquid droplets in the recess groove 55 can be mixed with the nucleic acid amplification reagent 25 in the nucleic acid amplification tube 20. .

In this way, the recess groove 55 is provided in the plunger 50, so that the recess groove 55 is immersed in the illusion buffer 15d accommodated in the chamber 10d of the extraction cartridge 10, and then the extraction cartridge ( A certain amount of the illusion buffer 15d accommodated in the chamber 10d of 10 is formed in the form of droplets in the recess groove 55 due to surface tension, and the illusion buffer 15d is quantitatively dispensed to produce nucleic acids. It can be precisely extracted and amplified.

The plunger lifting unit elevates the plunger (50). The plunger lifting part includes a support bracket 61 for the plunger, a screw shaft 65 for the plunger, and a drive motor 67 for the plunger.

The plunger support bracket 61 has an arm shape and is provided with a plunger mounting portion 63 to which the plunger 50 is attached and detached. Accordingly, the plunger 50 is suspended and supported on the plunger support bracket 61.

The plunger screw shaft 65 has a rod shape of a certain length, and a male thread is formed on the outer periphery of the plunger screw shaft 65 to be screwed to the plunger support bracket 61. The plunger screw shaft 65 is rotatably coupled to the plunger support bracket 61 to raise and lower the plunger support bracket 61.

The plunger drive motor 67 is directly connected to the plunger screw shaft 65 through a coupling (not shown), and rotates the plunger screw shaft 65 forward and backward. As the plunger screw shaft 65 rotates forward and backward, the plunger support bracket 61 moves up and down toward the extraction cartridge 10, and the plunger 50 disposed above the extraction cartridge 10 moves the extraction cartridge 10. ) is raised to the chambers 10a, 10b, 10c, and 10d.

The magnetic rod 70 has a rod shape of a certain length and is inserted into and extracted from the plunger 50. The magnetic rod 70 is disposed above the plunger 50. The tip of the magnetic rod 70, for example, at one end of the magnetic rod 70, is provided with a magnetic material 75 that generates a magnetic force for magnetically coupling to the magnetic bead 5. Accordingly, when the magnetic rod 70 is inserted into the plunger 50, the magnetic beads 5 accommodated in the chambers 10a, 10b, 10c, and 10d of the extraction cartridge 10 are moved to the plunger by the magnetic force of the magnetic rod 70. It is attached (binding) to the surface of (50). Conversely, when the magnetic rod 70 is pulled out from the plunger 50, the magnetic bead 5 attached to the surface of the plunger 50 is separated from the surface of the plunger 50 as the magnetic force of the magnetic rod 70 is lost. and is released into the buffers 15a, 15b, 15d and the solution 15c of the chambers 10a, 10b, 10c, and 10d of the extraction cartridge 10.

The rod lifting unit inserts and withdraws the magnetic rod 70 into and out of the plunger 50. The rod lifting unit includes a rod support bracket 81, a rod screw shaft 85, and a rod drive motor 87.

The rod support bracket 81 has an arm shape and suspends and supports the magnetic rod 70.

The rod screw shaft 85 has a rod shape of a certain length, and a male thread is formed on the outer periphery of the rod screw shaft 85 to be screwed to the rod support bracket 81. The rod screw shaft 85 is rotatably coupled to the rod support bracket 81 to raise and lower the rod support bracket 81.

The rod drive motor 87 is directly connected to the rod screw shaft 85 through a coupling (not shown) and rotates the rod screw shaft 85 forward and backward. As the rod screw shaft 85 rotates forward and backward, the rod support bracket 81 is raised and lowered toward the chambers 10a, 10b, 10c, and 10d of the extraction cartridge 10, and is disposed above the plunger 50. The magnetic rod 70 is raised and lowered inside the plunger 50.

In addition, the nucleic acid extraction and nucleic acid amplification device 1 according to an embodiment of the present invention further includes a reciprocating moving part that reciprocates the plunger 50 elevating part and the rod elevating part in the horizontal direction with respect to the elevating direction of the plunger 50. do.

The reciprocating portion includes a main support bracket 91, a main screw shaft 95, and a main drive motor 97.

The main support bracket 91 supports the plunger elevation portion and the rod elevation portion. Specifically, one end of the main support bracket 91 in this embodiment supports the plunger drive motor 67 and the rod drive motor 87, and at the same time includes the plunger screw shaft 65 and the rod screw. The shaft 85 penetrates and is screwed together. In addition, the main screw shaft 95 penetrates and is screwed to the other end of the main support bracket 91.

The main screw shaft 95 has a rod shape of a certain length, and a male screw that is screwed to the main support bracket 91 is formed on the outer circumference of the main screw shaft 95. The main screw shaft 95 is rotatably coupled to the main support bracket 91, and moves the main support bracket 91 horizontally in the ascending and descending direction of the plunger 50. For example, the plunger 50 and the magnetic The rod 70 is moved back and forth in the horizontal direction with respect to the lifting direction of the plunger 50.

The main drive motor 97 is directly connected to the main screw shaft 95 through a coupling (not shown) and rotates the main screw shaft 95 forward and backward. As the main screw shaft 95 rotates forward and backward, the main support bracket 91 reciprocates in the horizontal direction with respect to the ascending and descending direction of the plunger 50, thereby extracting the plunger 50 and the magnetic rod 70. 10) or placed above the chambers 10a, 10b, 10c, and 10d, or spaced apart from the chambers 10a, 10b, 10c, and 10d of the extraction cartridge 10. This main drive motor 97 is supported on the platform 30.

With this configuration, the process of separating, purifying and extracting nucleic acids from a sample, and amplifying nucleic acids using the nucleic acid extraction and nucleic acid amplification device 1 according to an embodiment of the present invention will be described as follows.

First, the process of separating, purifying, and extracting nucleic acids from the sample 15a will be described using FIGS. 4 to 8.

Prior to the explanation, hereinafter, for convenience of explanation, the chamber located on the rightmost side of the extraction cartridge 10 in FIG. 4 is referred to as the first chamber 10a, and is sequentially referred to as the second chamber 10b and the third chamber 10c. ), is called the fourth chamber (10d).

In order to separate, purify, and extract nucleic acids from a sample using the nucleic acid extraction and nucleic acid amplification device 1 according to the present invention, the magnetic beads 5 contained in the buffer 15b and the solution 15c are separated using a plunger ( The process of binding to the surface of the plunger 50 is representatively explained using the first chamber 10a as shown in FIG. 7, and the magnetic bead 5 attached to the surface of the plunger 50 is buffered. The process of releasing (15a, 15b, 15d) will be representatively explained using the second chamber (10b) as shown in FIG. 8.

First, it consists of a plurality of chambers (10a, 10b, 10c, 10d) containing magnetic beads containing the sample introduced for extraction, lysis buffer (15a), other buffers (15b, 15d), and solution (15c). The extraction cartridge 10 is seated on the cartridge seating portion 31, and at the same time, the nucleic acid amplification reagent tube 20 containing the nucleic acid amplification reagent 25 is seated on the tube seating portion 35.

Magnetic beads 5 are accommodated in the first chamber 10a, which accommodates a sample containing nucleic acid and a lysis buffer 15a. Then, the platform 30 is moved in a horizontal trajectory to cause mutual collision (bead beating) between the magnetic beads 5 and the dissolved nucleic acids in the sample and lysis buffer 15a, thereby causing the nucleic acids to hit the magnetic beads 5. Attach it.

Subsequently, as shown in FIG. 4, the main driving unit is driven to position the plunger 50 and the magnetic rod 70 above the first chamber 10a.

Then, as shown in (a) of FIG. 7, the rod drive motor 87 is driven so that the magnetic rod 70 is inserted into the plunger 50.

Next, the shaking motor is driven to move the extraction cartridge 10 mounted on the platform 30 in a horizontal trajectory.

At the same time, the plunger drive motor 67 and the rod drive motor 87 are synchronized and rotated forward and backward together so that the plunger 50 and the magnetic rod 70 are raised and lowered together in the first chamber 10a.

In this way, the sample and the lysis buffer 15a accommodated in the first chamber 10a are moved along the wall of the first chamber 10a along with numerous magnetic beads 5 by the horizontal trajectory movement of the platform 30. It flows and mixes at high speed, destroying the cell wall or mantle surrounding biological substances such as nucleic acids or antibodies contained in the sample, and biological substances such as nucleic acids and antibodies are separated from the sample and then attached to the magnetic beads (5).

In addition, the magnetic beads 5 on which biological substances such as nucleic acids and antibodies are adsorbed are attached to the plunger 50 by the magnetic force of the magnetic rod 70 inserted into the plunger 50, as shown in (b) of FIG. 7. It is attached to the surface, for example, of the plunger 50 located in the first chamber 10a. At this time, the sample flowing in the vertical direction along the wall of the first chamber 10a by the magnetic force of the magnetic rod 70 that rises and falls in the chamber together with the plunger 50 and the magnetic beads mixed in the lysis buffer 15a (5) can be more evenly attached to the surface of the plunger (50).

Then, when the attachment of the magnetic bead 5 to the surface of the plunger 50 is completed, the plunger drive motor 67 and the rod drive motor 87 are driven to connect the plunger 50 and the magnetic rod 70. Together, they are spaced apart from the first chamber 10a.

As a result, the process of nucleic acid being separated from the sample and adsorbed to the magnetic beads 5 is completed.

Below, the process of washing and eluting the magnetic beads 5 to which nucleic acids are adsorbed, separated from the sample and the lysis buffer 15a as described above, will be described.

Prior to explanation, the washing process of the magnetic beads 5 to which nucleic acids are adsorbed is performed in the second chamber 10b mounted on the cartridge seating portion 31.

First, the main drive motor 97 is driven to the upper part of the second chamber 10b, which accommodates the washing buffer 15b and is seated on the cartridge seating portion 31 of the platform 30, as shown in FIG. 8. As shown in (a), the plunger 50 and the magnetic rod 70 with the magnetic bead 5 attached to the surface of the plunger 50 are moved.

Next, the plunger drive motor 67 and the rod drive motor 87 are driven to accommodate the plunger 50 and the magnetic rod 70 together in the second chamber 10b.

Next, as shown in (b) of FIG. 8, the rod driving motor 87 is driven so that the magnetic rod 70 is spaced apart from the plunger 50.

As a result, the magnetic beads 5 to which nucleic acids are adsorbed can be separated from the surface of the plunger 50 as the magnetic force of the magnetic rod 70 is lost, and can be dispersed into the washing buffer 15b of the second chamber 10b. You can.

Then, the shaking motor is driven to move the extraction cartridge 10 along a horizontal trajectory to separate the magnetic bead 5 accommodated in the second chamber 10b from the plunger 50 and wash the magnetic bead 5. . At this time, foreign substances adsorbed together with the nucleic acid on the magnetic beads 5 are washed by the washing buffer 15b, are separated from the magnetic beads 5, and remain in the washing buffer 15b of the second chamber 10b.

When washing is completed, the plunger 50 and the magnetic rod 70 are accommodated in the second chamber 10b with the magnetic rod 70 inserted into the plunger 50 again, and the magnetic force of the magnetic rod 70 is applied. The washed magnetic beads 5 are attached to the surface of the plunger 50.

In addition, the process of attaching the magnetic beads 5 to the surface of the plunger 50 from the solution 15c containing the magnetic beads contained in the third chamber 10c is the same as the washing process described above, and will be described below. The detailed explanation is omitted.

Meanwhile, the magnetic beads 5 recovered through the third chamber 10c are moved to the fourth chamber 10d where the illusion buffer 15c (elusion buffer) is accommodated, and the final process, the illusion process, is performed.

By driving the main drive motor 97, the plunger 50 and the magnetic rod 70 with the washed magnetic beads 5 attached to the surface of the plunger 50 are placed in the fourth chamber 10d where the illusion buffer 15c is accommodated. ) moves upward.

Next, the plunger drive motor 67 and the rod drive motor 87 are driven to accommodate the plunger 50 and the magnetic rod 70 together in the fourth chamber 10d.

Next, the rod driving motor 87 is driven so that the magnetic rod 70 is spaced apart from the plunger 50.

At this time, as the magnetic rod 70 moves away from the plunger 50, the magnetic force of the magnetic rod 70 is lost, and the magnetic bead 5 to which the nucleic acid is adsorbed can be separated from the surface of the plunger 50, It may be distributed to the illusion buffer 15c of the fourth chamber 10d.

Then, the shaking motor is driven to move the extraction cartridge 10 along a horizontal trajectory to disperse the magnetic beads 5 from the surface of the plunger 50 into the illusion buffer 15d accommodated in the fourth chamber 10d. The nucleic acid adsorbed on the beads (5) is released and the nucleic acids are separated from the magnetic beads (5).

When the illusion is completed, although not shown, the main drive unit is driven to position the plunger 50 and the magnetic rod 70 above the fourth chamber 10d.

Then, the rod drive motor 87 is driven so that the magnetic rod 70 is inserted into the plunger 50.

Next, the plunger drive motor 67 and the rod drive motor 87 are driven so that the plunger 50 and the magnetic rod 70 are accommodated together in the fourth chamber 10d.

At this time, the pure magnetic beads 5 from which the nucleic acid has been separated by the illusion buffer 15d are attached to the surface of one end of the plunger 50 by the magnetic force of the magnetic rod 70.

Then, the plunger drive motor 67 and the rod drive motor 87 are driven so that the plunger 50 and the magnetic rod 70 are spaced apart from the fourth chamber 10d.

As a result, pure nucleic acid and the illusion buffer (15d) remain in the fourth chamber (10d), and the purification, separation, and extraction process of nucleic acid as an extract is completed.

Hereinafter, the process of transferring the nucleic acid to the illusion buffer 15c containing the extracted nucleic acid through the extraction cartridge 10 will be described.

After the nucleic acid extraction process is completed, with the illusion buffer 15d containing the nucleic acid contained in the fourth chamber 10d, the drive motor 67 for the plunger is driven to drive the recess groove 55 of the plunger 50. ) is lowered so that the plunger 50 is immersed in the illusion buffer 15d and accommodated in the fourth chamber 10d.

When immersion of the plunger 50 is completed, the plunger drive motor 67 is driven so that the plunger 50 rises, thereby separating the plunger 50 from the fourth chamber 10d, as shown in FIG. 5. At this time. At the tip of the plunger 50, a certain amount of the illusion buffer 15d containing the nucleic acid contained in the fourth chamber 10d is deposited in the recess groove 55 in the form of droplets due to surface tension.

Next, the main driver is driven to position the plunger 50 above the nucleic acid amplification reagent tube 20 mounted on the tube seating portion 35.

Then, the plunger 50 is driven so that the tip of the plunger 50 is immersed in the nucleic acid amplification reagent 25 accommodated inside the nucleic acid amplification reagent tube 20, so that the tip of the plunger 50 is immersed in the nucleic acid amplification reagent 25 as shown in FIG. 6. The plunger 50 is lowered so that the illusion buffer 15d containing the nucleic acid formed in the form of a droplet in the access groove 55 is immersed in the nucleic acid amplification reagent 25.

When immersion of the plunger 50 is completed, the plunger drive motor 67 is driven so that the plunger 50 rises, thereby separating the plunger 50 from the nucleic acid amplification reagent tube 20. At this time, the mixture of the nucleic acid amplification reagent 25 and the illusion buffer 15d containing the nucleic acid contained in the nucleic acid amplification reagent tube 20 is placed on the tip of the plunger 50 by surface tension in the recess groove ( 55), a certain amount of droplets are discharged from the nucleic acid amplification reagent tube 20, making it possible to quantitatively transfer the illusion buffer 15d containing nucleic acids. The nucleic acid amplification reagent 25 including the illusion buffer 15c may be amplified by increasing the temperature of the nucleic acid amplification reagent 25 in the heating module 36 located at the bottom of the nucleic acid amplification reagent tube seating portion 35. At this time, the heating module 36 can increase the temperature using a Peltier element.

Of course, the nucleic acid amplification tube 20 may be separated before nucleic acid amplification and performed in a separate nucleic acid amplification device (PCR Instrument).

In addition, since the nucleic acid purification, separation, and extraction process and the nucleic acid delivery process are performed in one device without exposure to the outside, contamination of the nucleic acid to be amplified is prevented and the nucleic acid is safely separated, purified, and extracted. It is possible to amplify the extract, purification and extraction process, and automation of the nucleic acid amplification process. In order to amplify the nucleic acid, the operator does not need to dispense and transport the nucleic acid to be amplified, thereby eliminating the need for the nucleic acid amplification process. It is possible to prevent errors or contamination during dispensing and transfer.

Meanwhile, in the above-described embodiment, one extraction cartridge composed of a plurality of chambers arranged in a row, one plunger, and one magnetic rod are described as being provided, but the present invention is not limited thereto, and the plurality of extraction cartridges are provided in a row. and are arranged in a row in a row, and correspondingly, plungers and magnetic rods are provided in quantities corresponding to the quantity arranged in the row or column direction of a plurality of extraction cartridges, allowing for simultaneous separation, purification, and extraction of a large number of samples. , a nucleic acid amplification process may be performed.

In addition, in the above-described embodiment, the support bracket for the plunger and the screw shaft for the plunger are screwed together, and the plunger is shown to be raised and lowered by driving the drive motor for the plunger. However, this is not limited to this, and the linear reciprocating motion of the plunger is not limited thereto. The plunger can be raised and lowered by directly connecting to a motor, or the plunger can be raised and lowered by directly connecting to the cylinder rod of a fluid pressure cylinder that extends and shortens the plunger.

In the above-described embodiment, the support bracket for the rod and the screw shaft for the rod are screwed together, and the magnetic rod is shown to be raised and lowered by driving the rod drive motor. However, this is not limited to this, and the magnetic rod is moved in a linear reciprocating motion. The magnetic rod can be raised and lowered by directly connecting to a linear motor that extends or shortens the magnetic rod, or it can be directly connected to the cylinder rod of a fluid pressure cylinder that extends and shortens the magnetic rod.

In addition, in the above-described embodiment, the main support bracket and the main screw shaft are screwed together, and the plunger lifting part and the rod lifting part are shown to be raised and lowered by the driving of the main drive motor, but the present invention is not limited to this, and the plunger lifting part and the rod are not limited to this. The lifting part is directly connected to a linear motor that moves in a linear motion to raise and lower the plunger lifting part and the rod lifting part, or the plunger lifting part and the rod lifting part are raised and lowering by directly connecting to the cylinder rod of a fluid pressure cylinder that extends and shortens the length of the plunger lifting part and the rod lifting part. Wealth can also be promoted.

1: Nucleic acid extraction and nucleic acid delivery device
5: magnetic bead
10: Extraction cartridge
10a, 10b, 10c, 10d: Chamber
20: Nucleic acid amplification reagent tube
30: platform
31: Cartridge seating portion
35: Tube seating portion
36: Heating module
50: Plunger
55: Recess groove
61: Support bracket for plunger
63: Plunger mounting part
65: Screw shaft for plunger
67: Drive motor for plunger
70: magnetic rod
75: magnetic material
81: Support bracket for rod
85: Screw shaft for rod
87: Driving motor for rod
90: Reciprocating moving part
91: Main support bracket
95: Main screw shaft
97: Main driving motor

Claims (7)

It has a cartridge seating portion on which a plurality of extraction cartridges containing a mixed sample and a buffer are seated, and a tube seating portion provided adjacent to the cartridge seating portion on which a plurality of nucleic acid amplification reagent tubes containing a nucleic acid amplification reagent (PCR reaction) are seated. platform;
a plunger that raises and lowers to selectively insert or extract one of the plurality of extraction cartridges mounted on the cartridge seating portion and the plurality of nucleic acid amplification reagent tubes mounted on the tube seating portion;
a plunger lifting part that has a plunger mounting part to which the plunger is attached and detached, and that raises and lowers the plunger;
a magnetic rod that moves up and down to be inserted into or extracted from the plunger, and attaches a magnetic bead accommodated in the extraction cartridge to the plunger or separates it from the plunger;
A nucleic acid extraction and nucleic acid delivery device comprising a rod lifting unit that elevates a commercial magnetic rod. According to paragraph 1,
The plunger,
A nucleic acid extraction and nucleic acid delivery device comprising a recess groove formed in the tip of the plunger into which a certain amount of droplets of the buffer contained in the extraction cartridge are formed. According to paragraph 2,
A nucleic acid extraction and nucleic acid delivery device, wherein the recess groove has a semi-spherical or partially spherical and polygonal cross-sectional shape. According to paragraph 1,
A nucleic acid extraction and nucleic acid delivery device further comprising a heating module provided on the tube mounting portion and heating the nucleic acid amplification reagent tube mounted on the tube mounting portion. According to paragraph 1,
The plunger lifting part,
a support bracket for the plunger supporting the plunger mounting portion;
a plunger screw shaft rotatably coupled to the plunger support bracket to raise and lower the plunger support bracket;
A nucleic acid extraction and nucleic acid delivery device comprising a drive motor for the plunger that rotates the screw shaft for the plunger. According to paragraph 1,
The load lifting unit,
a rod support bracket supporting the magnetic rod;
a rod screw shaft rotatably coupled to the rod support bracket to elevate and lower the rod support bracket;
A nucleic acid extraction and nucleic acid delivery device comprising a rod driving motor that rotates the rod screw shaft. According to paragraph 1,
It further includes a reciprocating moving part that reciprocates the plunger lifting part and the rod lifting part in a horizontal direction with respect to the lifting direction of the plunger,
The reciprocating moving part,
a main support bracket supporting the plunger lifting part and the rod lifting part;
a main screw shaft rotatably coupled to the main support bracket and reciprocating the main support bracket in a horizontal direction with respect to the ascending and descending direction of the plunger;
A nucleic acid extraction and nucleic acid delivery device comprising a main driving motor that rotates the main screw shaft.