KR102317030B1 - Nucleic acid extraction device and nucleic acid extraction method - Google Patents

Abstract

The present invention relates to a nucleic acid extraction apparatus and a nucleic acid extraction method, in which a sample including magnetic nanoparticles and pathogens is stored, and a concentration unit for separating and concentrating pathogens from the sample, and the concentration unit is rotatably mounted and discharged from the concentration unit A collection unit for processing a sample, a storage unit connected to the collection unit and capable of storing one or more buffers; Including the refining unit, the cost and time required for the diagnostic test can be saved.

Description

Nucleic acid extraction apparatus and nucleic acid extraction method

The present invention relates to a nucleic acid extracting apparatus and a nucleic acid extraction method, and to a nucleic acid extracting apparatus and a nucleic acid extraction method capable of simultaneously concentrating multiple pathogens present in a sample to remove diagnostic inhibitors, and purifying and extracting nucleic acids from the concentrated pathogen. it's about

In general, hospitals incubate a sample of a patient collected for at least 12 hours to detect a low-concentration pathogen. After that, the cultured sample is used for diagnostic testing after the pathogen is separated through a centrifuge. However, this method takes a long time and it is impossible to completely remove the inhibitors of molecular diagnostics, so it adversely affects the detection sensitivity based on molecular diagnostics. In addition, the amount used for molecular diagnosis is about 2 μL compared to the collected sample (2-5 mL).

In addition, there is a problem in that the installation cost increases because the device for concentrating the pathogen, the device for purifying the nucleic acid from the concentrated pathogen, and the device for extracting the nucleic acid are separately installed. Therefore, there is a need to improve it.

Background art of the present invention is disclosed in Korean Patent Application Laid-Open No. 2013-0101606 (published on September 16, 2013, title of the invention: ultrafast nucleic acid extraction apparatus, and nucleic acid extraction method using the same).

The present invention has been devised to improve the above problems, and a nucleic acid extraction apparatus and nucleic acid extraction method capable of simultaneously concentrating multiple pathogens present in a sample to remove diagnostic inhibitors, and purifying and extracting nucleic acids from the concentrated pathogens. Its purpose is to provide

A nucleic acid extraction apparatus according to the present invention includes: a concentration unit for storing a sample including magnetic nanoparticles and a pathogen, and separating and concentrating the pathogen from the sample; a collecting unit which is rotatably mounted to the concentrator and processes the sample discharged from the concentrator; a storage unit connected to the collection unit and capable of storing one or more buffers; and a purification unit connected to the storage unit, receiving the concentrated pathogen supplied from the concentrating unit, purifying it into nucleic acid, and discharging the purified nucleic acid.

The concentrating unit includes a concentrating tube in which a sample containing magnetic nanoparticles and pathogens is stored; a concentration insert extending downward of the enrichment tube, inserted into the collecting unit, and rotatable in the collecting unit; and an enrichment outlet formed in the enrichment insertion unit and discharging a sample containing magnetic nanoparticles and pathogens.

The concentrating unit is detachable from the concentrating tube portion, and the concentrating magnetic unit providing a magnetic force; characterized in that it further comprises.

The collecting unit includes: a collecting receiving unit on which the concentrating unit is rotatably seated; a collection guide unit formed in the collection receiving unit and guiding the sample discharged from the enrichment unit; and a collection storage unit disposed below the collection receiving unit and storing the sample discharged from the concentrating unit.

The collection part is formed in the collection seat part, and a collection passage part for guiding the concentrated pathogen discharged from the concentration part to the purification part; characterized in that it further comprises.

The storage unit is connected to the collection unit, and a storage space for storing one or more buffers is formed; a storage passage for guiding the buffer stored in the storage space to the purification unit; and a storage valve unit for opening and closing the storage passage unit.

The storage valve unit is characterized in that the rotary valve method is applicable.

The purification unit is connected to the lower portion of the storage unit, the pathogen concentrated in the concentration unit is introduced, and a buffer is introduced into the storage unit to purify the nucleic acid; and a tablet discharging unit formed under the tablet cylinder and discharging the purified nucleic acid from the tablet cylinder.

The tablet unit may further include a tablet magnetic force unit that is detachable from the tablet cylinder and provides a magnetic force.

A nucleic acid extraction method according to the present invention comprises: a sample preparation step of mixing magnetic nanoparticles with a sample containing a pathogen; A sample supply step of supplying a sample including pathogens and magnetic nanoparticles to the concentration unit; a concentration step in which pathogens and magnetic nanoparticles are separated from the sample and concentrated by the concentration unit; a transfer step of transferring the pathogen concentrated in the concentrating unit to the refining unit; an input step of selectively injecting the buffer stored in the storage into the refining unit; an extraction step of extracting nucleic acids from the pathogen transferred to the purification unit; and a purification step of purifying the extracted nucleic acid.

The magnetic nanoparticles are characterized in that by coating probes that bind to a specific layer on the surface of the pathogen, Gram-positive bacteria and Gram-negative bacteria can be simultaneously concentrated in the sample.

The concentration step is characterized in that the magnetic nanoparticles are separated from the sample by the magnetic force provided to the concentration part and concentrated.

The transferring step is characterized in that after removing the magnetic force provided to the concentrating unit and providing the magnetic force to the refining unit, the concentrated pathogen is transferred to the refining unit.

The nucleic acid extraction apparatus and nucleic acid extraction method according to the present invention are required for diagnostic testing because a low-concentration pathogen present in a large-capacity sample can be rapidly separated from the sample using magnetism, and then concentrated to purify and isolate the nucleic acid in the purification unit. can save cost and time.

Since the nucleic acid extraction apparatus and the nucleic acid extraction method according to the present invention use magnetic nanoparticles coated with probes that bind to a specific layer on the surface of the pathogen, multiple concentration of pathogens in the sample is possible.

In the nucleic acid extraction apparatus and the nucleic acid extraction method according to the present invention, all buffers necessary for the storage unit integrally provided in the nucleic acid extraction apparatus can be stored, so that workability can be improved.

The nucleic acid extraction apparatus and nucleic acid extraction method according to the present invention can make pathogens in a sample in a range not detected by molecular diagnosis into a detectable range.

1 is a diagram schematically showing a nucleic acid extracting apparatus according to an embodiment of the present invention.
2 is a view schematically showing a concentrator according to an embodiment of the present invention.
3 is a diagram schematically showing a collection unit according to an embodiment of the present invention.
4 is a diagram schematically illustrating a storage unit according to an embodiment of the present invention.
5 is a view schematically showing a tablet unit according to an embodiment of the present invention.
6 is a flowchart schematically illustrating a nucleic acid extraction method according to an embodiment of the present invention.

Hereinafter, embodiments of a nucleic acid extraction apparatus and a nucleic acid extraction method according to the present invention will be described with reference to the accompanying drawings. In this process, the thickness of the lines or the size of the components shown in the drawings may be exaggerated for clarity and convenience of explanation. In addition, the terms to be described later are terms defined in consideration of functions in the present invention, which may vary according to intentions or customs of users and operators. Therefore, definitions of these terms should be made based on the content throughout this specification.

1 is a diagram schematically showing a nucleic acid extracting apparatus according to an embodiment of the present invention. Referring to FIG. 1 , the nucleic acid extraction apparatus 1 according to an embodiment of the present invention includes a concentrating unit 10 , a collecting unit 20 , a storage unit 30 , and a purification unit 40 . .

A sample including magnetic nanoparticles and a pathogen is stored in the concentration unit 10, and the pathogen is separated from the sample and concentrated. The collecting unit 20 on which the concentrating unit 10 is rotatably mounted processes the sample discharged from the concentrating unit 10 . The storage unit 30 is connected to the collection unit 20 and stores one or more buffers. The purification unit 40 is connected to the storage unit 30, receives the concentrated pathogen supplied from the concentration unit 10, purifies it into nucleic acid, and discharges the purified nucleic acid.

The nucleic acid extracting apparatus 1 having the above configuration simultaneously multi-concentrates low-concentration pathogens present in the sample (2-10 mL) to remove diagnostic inhibitors, and purifies and extracts nucleic acids from the concentrated pathogens. can Since the nucleic acid extraction device 1 provides an integrated cartridge in which the above components are combined, all pathogens are used for molecular diagnostic testing, thereby improving detection sensitivity and enabling rapid diagnostic testing.

On the other hand, in the nucleic acid extraction apparatus 1, the collecting unit 20, the storage unit 30, and the purification unit 40 are integrally formed, and the concentrator 10 is rotatably mounted to the collecting unit 20. can be The nucleic acid extracting device 1 is produced by a 3D printer or injection molding, and when the bacterial pathogen is concentrated in the concentration unit 10, the concentrated pathogen is transferred to the purification unit 40, and a washing process is performed. Including nucleic acid purification and extraction process.

In this case, nucleic acid means an acidic substance in the nucleus, and DNA and RNA are called nucleic acids. Nucleic acids are composed of numerous nucleotides linked together, and these nucleotides are the basic units of nucleic acids and consist of three parts: phosphate, sugar, and base.

And, DNA is a polymer complex composed of four nucleotides A, G, C, and T. It is a negatively charged biopolymer material that contains the genetic information of living things. Therefore, in order to analyze the composition, content, and properties of genes, high-purity DNA must first be isolated. Nucleic acids are released as pathogens, which are bacteria, are dissolved, and the surface of the nucleic acids is negatively charged.

2 is a view schematically showing a concentrator according to an embodiment of the present invention. Referring to FIG. 2 , the concentrating unit 10 according to an embodiment of the present invention includes a concentrating tube 11 , a concentrating inserting unit 13 , and a concentrating outlet 14 .

The concentrating tube 11 stores a sample containing magnetic nanoparticles and pathogens. As an example, the enrichment cylinder part 11 may have a cylindrical shape and an upper open shape. The sample input to the concentrating tube 11 uses magnetic nanoparticles coated with probes such as vancomycin and polymyxin that bind to a specific layer on the surface of a pathogen rather than a specific antigen. . Due to this, Gram-positive bacteria and Gram-negative bacteria can be simultaneously concentrated in the sample.

On the other hand, the enrichment cover portion 12 may open and close the opened region of the enrichment tube portion (11). For this reason, the sample containing the magnetic nanoparticles and the pathogen may be introduced into the area opened through the concentration cover part 12 .

The enrichment insertion unit 13 extends downward of the enrichment tube 11 , is inserted into the collecting unit 20 , and is rotatable in the collecting unit 20 . For example, the enrichment insert 13 may have a smaller diameter than the enrichment tube 11 so that the bottom surface of the enrichment tube 11 may be seated on the collecting unit 20 .

The enrichment outlet 14 is formed in the enrichment insertion unit 13, and discharges a sample containing magnetic nanoparticles and pathogens. For example, the enrichment outlet 14 may be a hole or a pipe formed in the lower portion of the enrichment inserting unit 13 .

The concentrating unit 10 according to an embodiment of the present invention may further include a concentrating magnetic force unit 15 that is detachable from the concentrating tube unit 11 and provides a magnetic force. As an example, the concentrating magnetic force unit 15 may be a permanent magnet or an electromagnet, and may be mounted on the outer circumferential surface of the concentrating tube unit 11 . The pathogen connected to the magnetic nanoparticles by the concentrating magnetic unit 15 may be fixed to the inside of the concentrating tube 11 and concentrated.

3 is a diagram schematically showing a collection unit according to an embodiment of the present invention. Referring to FIG. 3 , the collection unit 20 according to an embodiment of the present invention includes a collection seating unit 21 , a collection guide unit 22 , and a collection storage unit 23 .

The collection seating part 21 is seated so that the enrichment part 10 is rotatable. As an example, the collection receiving part 21 may have a collecting groove 211 formed therein so that the concentration inserting part 13 is inserted in the central portion to be rotatable. Due to this, the edge of the concentrating tube 11 may be seated on the edge of the collection seating part 21 .

The collection guide 22 is formed on the collection seat 21 , and guides the sample discharged from the concentrator 10 . As an example, the collection guide 22 may be a hole or a pipe formed in the collection seat 21 , and the collection guide 22 and the concentration outlet 14 are connected by the rotation of the concentration inserting part 13 . When communicating, the sample excluding the pathogen connected to the magnetic nanoparticles stored in the enrichment tube 11 and the enrichment insertion unit 13 may be discharged.

The collection storage unit 23 is disposed below the collection seating unit 21 , and stores the sample discharged from the concentration unit 10 . For example, the collection storage unit 23 is detachable from the lower portion of the collection mounting unit 21 , and when a sample is stored in the collection storage unit 23 , the collection storage unit 23 is separated from the collection mounting unit 21 to After discarding the sample, the emptied collection storage unit 23 may be remounted to the collection mounting unit 21 .

The collection unit 20 according to an embodiment of the present invention may further include a collection passage unit 24 . The collection passage part 24 is formed in the collection seating part 21 , and guides the concentrated pathogen discharged from the concentration part 10 to the purification part 40 . As an example, when the collection flow path part 24 and the concentration outlet part 14 communicate with the rotation of the concentration insertion part 13, the concentrated pathogens stored in the concentration tube part 11 and the concentration insert part 13 can be guided. have. The collection passage part 24 may pass through the storage part 30 and communicate with the purification part 40 .

4 is a diagram schematically illustrating a storage unit according to an embodiment of the present invention. Referring to FIG. 4 , the storage unit 30 according to an embodiment of the present invention includes a storage cylinder unit 31 , a storage flow path unit 32 , and a storage valve unit 33 .

The storage container 31 is connected to the collection unit 20 and a storage space 313 for storing one or more buffers is formed. For example, the storage barrel 31 may include a storage shaft portion 311 having a cylindrical shape, and a storage plate portion 312 in a disk shape extending to the outer circumferential surface of the storage shaft portion 311 and having a vertical thickness. In addition, a plurality of storage spaces 313 are formed in the circumferential direction of the storage plate unit 312 to store various types of buffers.

The storage flow path unit 32 guides the buffer stored in the storage space 313 to the purification unit 40 . For example, the storage passage 32 may have an upper end in communication with each storage space 313 , and a lower end communicating with the storage shaft 311 .

The storage valve part 33 opens and closes the storage passage part 32 to selectively discharge the buffer stored in the storage space 313 . This storage valve part 33 is applicable to the rotary valve method. As an example, the storage valve part 33 includes a valve shaft part 331 passing through the storage shaft part 311, a valve operation part 332 protruding laterally from the upper part of the valve shaft part 331 and operable, and a valve shaft part ( It may include a valve flow path part 333 formed in 331 and selectively communicating with the storage flow path part 32 . For this reason, when the valve shaft part 331 is rotated by manual or automatic operation of the valve operation part 332, the valve flow path part 333 is selectively communicated with any one of the plurality of storage flow path parts 32 to convert the buffer into the purification unit ( 40) can be guided.

5 is a view schematically showing a tablet unit according to an embodiment of the present invention. Referring to FIG. 5 , the tablet unit 40 according to an embodiment of the present invention includes a tablet cylinder unit 41 and a tablet discharge unit 42 .

The purification barrel 41 is connected to the lower portion of the storage unit 30 , the pathogen concentrated in the concentration unit 10 is introduced, and a buffer is introduced in the storage unit 30 to purify the nucleic acid. For example, the tablet cylinder part 41 may be connected to the lower end of the storage shaft part 311 , communicate with the collection passage part 24 , and have a shape in which the inner diameter decreases toward the bottom.

The tablet discharge unit 42 is formed at the lower portion of the tablet cylinder unit 41 , and discharges the purified nucleic acid from the tablet cylinder unit 41 . For example, the tablet discharge unit 42 may include a discharge pipe unit 421 connected to the lower portion of the tablet cylinder unit 41 and a discharge valve unit 422 for opening and closing the discharge pipe unit 421 .

The refining unit 40 according to an embodiment of the present invention may further include a refining magnetic force unit 43 . The tablet magnetic force part 43 is detachable from the tablet cylinder part 41 and provides a magnetic force. For example, the tablet magnetic force unit 43 may be a permanent magnet or an electromagnet, and may be mounted on the outer peripheral surface of the tablet cylinder unit 41 .

6 is a flowchart schematically illustrating a nucleic acid extraction method according to an embodiment of the present invention. A nucleic acid extraction method according to an embodiment of the present invention will be described with reference to FIG. 6 as follows.

First, it has a sample preparation step of mixing the magnetic nanoparticles to the sample containing the pathogen (S10). These magnetic nanoparticles are coated with probes such as vancomycin and polymyxin that bind to a specific layer on the surface of a pathogen rather than a specific antigen. The magnetic nanoparticles can simultaneously concentrate Gram-positive bacteria and Gram-negative bacteria in the sample.

When the sample preparation is completed, a sample supply step of supplying a sample including pathogens and magnetic nanoparticles to the concentrator 10 is performed (S20). For example, when the enrichment cover 12 opens the enrichment tube 11 , a sample including pathogens and magnetic nanoparticles is supplied to the enrichment tube 11 . In addition, the sample may be supplied through a supply passage connected to the concentrating tube 11 .

When the sample is supplied to the concentration unit 10, the concentration unit 10 has a concentration step of separating the pathogen and the magnetic nanoparticles from the sample and concentrating (S30). That is, the magnetic nanoparticles and pathogens connected to the magnetic nanoparticles are separated from the sample and concentrated by the magnetic force provided to the concentrator 10 . In this concentration step, the sample separated from the pathogen may be collected by the collecting unit 20 and then discarded.

When the pathogen is concentrated through the concentration step, there is a transfer step of transferring the pathogen concentrated in the concentration unit 10 to the purification unit 40 (S40). In the transfer step, the magnetic force provided to the concentrating unit 10 is blocked and the magnetic force is provided to the refining unit 40 , and then the concentrated pathogen is transferred to the refining unit 40 .

That is, when the magnetic force provided to the enrichment unit 10 is blocked, the magnetic nanoparticles and pathogens concentrated in the enrichment unit 10 are in a state in which movement is possible. Then, using 1 mL of phosphate buffered saline (PBS), the concentrated pathogens and magnetic nanoparticles are moved in the direction of the purification unit 40 . At this time, by rotating the enrichment insertion part 13, the enrichment outlet part 14 and the collection passage part 24 communicate,

PBS is a phosphate buffer and is the most used buffer in biological experiments and is used as a buffer solution. A buffer solution is a solution that tries to maintain a constant hydrogen ion concentration without being affected by any acid or base added from the outside.

In addition, in the transfer step, a solution other than phosphate buffered saline may be used within the technical concept of supplying a washing solution after blocking the supply of magnetic force in the concentrating unit 10 to transfer the concentrated pathogen to the refining unit 40 .

When the concentrated pathogen is transferred to the purification unit 40, an input step of selectively inputting the buffer stored in the storage unit 30 to the purification unit 40 is performed (S50).

That is, various types of buffers are respectively stored in the plurality of storage spaces 313 formed in the storage barrel 31 , and the buffers are selectively introduced into the purification unit 40 by the operation of the storage valve unit 33 . can

For example, 100 μL of Lysis buffer and 100 μL of Binding buffer may be stored in the plurality of storage spaces 313 .

Lysis buffer is a buffer used to disrupt open cells for use in molecular biology experiments that analyze cellular compounds. Most lysis buffers contain salts to control the acidity and osmotic pressure of the lysate. Sometimes detergents (eg Triton X-100 or SDS) can be added to dissolve the membrane structure, and these lysis buffers can be used in both animal and plant tissue cells. Lysis buffer is used to separate nucleic acids by dissolving pathogens bound to magnetic nanoparticles.

The binding buffer functions as a buffer to allow the magnetic silica particles to be supplied to the purification barrel 41 and nucleic acids to bind thereafter.

In addition, 10 μL of Proteinase K and 5 μL of RNase A may be stored in the storage space 313 .

Proteinase K is a proteolytic enzyme as a reagent for increasing the purification efficiency of nucleic acids. Proteinase K removes histone proteins from DNA. Histone proteins are attached to DNA to maintain stability. So, if Proteinase K is added, the purity will decrease when DNA is extracted.

RNase (ribonuclease) is an enzyme used to cut RNA. The reason that RNase is used for DNA purification is that when DNA is usually extracted, RNA is also extracted in the same way, and what is needed is DNA.

When the buffer is added to the purification unit 40, an extraction step of extracting nucleic acids from the pathogen transferred to the purification unit 40 is performed (S60).

That is, after incubation while maintaining a constant temperature for 2 minutes at room temperature, 25 μL of magnetic silica particles and 200 μL of isopropanol are added to the purification unit 40, and vortex is performed for 5 minutes.

Nucleic acid is extracted by dissolving the concentrated pathogen in this process, and the extracted nucleic acid is electrostatically bonded to magnetic silica particles due to cation bridge. Since cationic crosslinking has a cation, magnetic silica particles and nucleic acids have anions, magnetic silica particles and nucleic acids are bonded to both sides of cationic crosslinking.

Isopropanol is an aliphatic saturated alcohol that is an isomer of propanol. The reason for vortexing is to mix well with the substance supplied for DNA purification.

Isopropanol and ethanol are the same alcohols and are used to increase the precipitation efficiency of DNA neutralized by salt.

And after stopping providing the magnetic force to the refining unit 40, and vortexing for 5 minutes, a magnetic force is provided to the lower portion of the refining cylinder 41 to discharge impurities.

When the nucleic acid extraction is completed, a purification step of purifying the extracted nucleic acid is performed (S70). That is, the process of injecting 350 μL of washing buffer through the storage unit 30 is repeated three times, and all impurities in the tablet cylinder unit 41 are discharged through the tablet discharge unit 42 . And the operation of removing the internal fluid in the tablet cylinder 41 is repeated three times.

Then, 100 μL of elution buffer is supplied to the tablet tub 41 through the storage 30 , and the magnetic force provided to the tablet tub 41 is blocked. Thereafter, after heating the purification cylinder 41 at a temperature of 65° C. with a heater for 5 minutes, the nucleic acids are separated from the magnetic silica particles. When the separated liquid sample is obtained, it is discharged to the outside through the purification discharge unit 42 .

The nucleic acid extraction apparatus and nucleic acid extraction method according to an embodiment of the present invention isolate a low-concentration pathogen present in a large-capacity sample from the sample using magnetism, and then concentrate it to purify and separate the nucleic acid in the purification unit 40 . Therefore, the cost and time required for the diagnostic test can be saved.

In addition, since the present invention uses magnetic nanoparticles coated with probes that bind to a specific layer on the pathogen surface, multiple concentration of pathogens in the sample is possible.

In addition, in the present invention, since all buffers necessary for the storage unit 30 integrally provided in the nucleic acid extraction apparatus 1 can be stored, workability can be improved.

Although the present invention has been described with reference to the embodiment shown in the drawings, this is merely exemplary, and it is understood that various modifications and equivalent other embodiments are possible by those of ordinary skill in the art. will understand Accordingly, the true technical protection scope of the present invention should be defined by the following claims.

10: enrichment unit 11: enrichment tube unit
12: enrichment cover part 13: enrichment insertion part
14: concentrating outlet 15: concentrating magnetic unit
20: collection unit 21: collection mounting unit
22: collection guide 23: collection storage unit
24: collection passage 30: storage
31: storage tube portion 32: storage passage portion
33: storage valve unit 40: refining unit
41: tablet cylinder part 42: tablet discharge part
43: refining magnetic force unit

Claims (13)

a concentration unit storing a sample including magnetic nanoparticles and pathogens, and separating and concentrating the pathogen from the sample;
a collecting unit which is rotatably mounted to the concentrator and processes the sample discharged from the concentrator;
a storage unit connected to the collection unit and capable of storing one or more buffers; and
a purification unit connected to the storage unit, receiving the concentrated pathogen supplied from the concentrating unit, purifying it into nucleic acid, and discharging the purified nucleic acid;
The collecting unit, the storage unit, and the refining unit are integrally formed,
the concentrator
a concentrating tube in which a sample containing magnetic nanoparticles and pathogens is stored;
a concentration insert extending downward of the enrichment tube, inserted into the collecting unit, and rotatable in the collecting unit; and
A nucleic acid extracting apparatus comprising a; is formed in the concentration insert, the concentration outlet for discharging the sample containing the magnetic nanoparticles and pathogens.
delete The method of claim 1, wherein the concentrating unit
A nucleic acid extracting device comprising a; detachable and detachable concentrator to the concentrating tube and providing a magnetic force.
According to claim 1, wherein the collection unit
a collection seating part on which the concentrating part is rotatably seated;
a collection guide unit formed in the collection receiving unit and guiding the sample discharged from the enrichment unit; and
A nucleic acid extracting apparatus comprising a; a collection storage unit disposed below the collection receiving part and storing the sample discharged from the concentration part.
5. The method of claim 4, wherein the collection unit
The nucleic acid extracting apparatus further comprising a; a collection passage formed in the collection receiving part and guiding the concentrated pathogen discharged from the concentration part to the purification part.
The method of claim 1, wherein the storage unit
a storage container connected to the collection unit and having a storage space for storing one or more buffers;
a storage passage for guiding the buffer stored in the storage space to the purification unit; and
A nucleic acid extraction device comprising a; a storage valve for opening and closing the storage passage.
7. The method of claim 6,
The storage valve unit is a nucleic acid extraction device, characterized in that the rotary valve method is applicable.
According to claim 1, wherein the purification unit
a purification barrel connected to the lower part of the storage unit, into which pathogens concentrated in the concentration unit are introduced, and a buffer is introduced from the storage unit to purify the nucleic acid; and
A nucleic acid extracting apparatus comprising a; a purification discharge unit formed at the lower portion of the purification barrel and discharging the purified nucleic acid from the purification barrel.
The method of claim 8, wherein the purification unit
The nucleic acid extracting apparatus further comprising a; a purification magnetic force unit detachable from the tablet barrel and providing a magnetic force.
A sample preparation step of mixing magnetic nanoparticles with a sample containing a pathogen;
A sample supply step of supplying a sample including pathogens and magnetic nanoparticles to the concentration unit;
a concentration step in which pathogens and magnetic nanoparticles are separated from the sample by the concentration unit and concentrated;
a transfer step of transferring the pathogen concentrated in the concentrating unit to the refining unit;
an input step of selectively injecting the buffer stored in the storage into the refining unit;
an extraction step of extracting nucleic acids from the pathogen transferred to the purification unit; and
Including; a purification step of purifying the extracted nucleic acid;
A nucleic acid extraction method, characterized in that the collecting unit for processing the sample discharged from the concentrating unit, the storage unit, and the purification unit are integrally formed, and the concentrator is rotatably mounted to the collecting unit.
11. The method of claim 10,
The magnetic nanoparticles are coated with probes that bind to a specific layer on the surface of the pathogen, so that Gram-positive and Gram-negative bacteria can be simultaneously concentrated in the sample.
11. The method of claim 10, wherein the concentration step
A nucleic acid extraction method, characterized in that magnetic nanoparticles are separated from the sample and concentrated by the magnetic force provided to the enrichment unit.
11. The method of claim 10, wherein the transferring step
A nucleic acid extraction method, characterized in that after removing the magnetic force provided to the enrichment unit and providing the magnetic force to the purification unit, the concentrated pathogen is transferred to the purification unit.

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