KR20210032641A - Nucleic acid extraction device and nucleic acid extraction method - Google Patents

Abstract

The present invention relates to a nucleic acid extraction device and a nucleic acid extraction method, wherein the nucleic acid extraction device comprises: a concentration unit which stores samples including magnetic nanoparticles and a pathogen, and separates and concentrates the pathogen from the sample; a collection unit which is mounted to enable the concentration unit to rotate, and treats the sample discharged from the concentration unit; a storage unit which is connected to the collection unit and is capable of storing one or more buffers; and a purification unit which is connected to the storage unit, and receives the concentrated pathogen supplied from the concentrating unit, purifies the same into nucleic acid, and discharges the purified nucleic acid. Therefore, the device of the present invention can save cost and time required for a diagnostic test.

Description

Nucleic acid extraction device and nucleic acid extraction method {NUCLEIC ACID EXTRACTION DEVICE AND NUCLEIC ACID EXTRACTION METHOD}

The present invention relates to a nucleic acid extraction device and a nucleic acid extraction method, wherein pathogens present in a sample are simultaneously multiplexed to remove diagnostic inhibitors, and a nucleic acid extraction device and a nucleic acid extraction method capable of purifying and extracting nucleic acids from the concentrated pathogens. About.

In general, hospitals incubate a sample of a patient for at least 12 hours to detect low concentrations of pathogens. After that, the cultured sample is used for diagnostic testing after separating the pathogen through a centrifuge. However, since this method takes a long time and cannot completely remove the inhibitory factor for molecular diagnosis, it adversely affects the detection sensitivity based on molecular diagnosis. In addition, compared to the collected sample (2-5 mL), the amount used for actual molecular diagnosis is about 2 μL, and since it is difficult to use all pathogens for diagnostic tests, a pathogen concentration technique is essential.

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

The background technology of the present invention is published in Korean Patent Application Publication No. 2013-0101606 (published on September 16, 2013, title of the invention: ultra-high-speed nucleic acid extraction device, and a nucleic acid extraction method using the same).

The present invention was conceived to improve the above problems, and a nucleic acid extraction device and a nucleic acid extraction method capable of removing diagnostic inhibitors by simultaneously concentrating pathogens present in a sample multiplexedly, and purifying and extracting nucleic acids from concentrated pathogens Its purpose is to provide.

The nucleic acid extraction apparatus according to the present invention includes: a concentration unit for storing a sample including magnetic nanoparticles and a pathogen, and for separating and concentrating the pathogen from the sample; A collection unit mounted so as to be rotatable in the concentrating unit and processing the sample discharged from the concentrating unit; 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 concentration unit, and purifying it with nucleic acid to discharge the purified nucleic acid.

The concentrating unit may include a condensing tube for storing a sample containing magnetic nanoparticles and pathogens; A concentration insertion portion extending downward of the concentration cylinder portion, inserted into the collection portion, and rotatable in the collection portion; And a concentration outlet formed in the concentration insertion portion and for discharging a sample containing magnetic nanoparticles and pathogens.

It characterized in that it further comprises a; the concentrating part is detachable to the concentrating cylinder part, and a magnetic concentrating part for providing a magnetic force.

The collection unit includes a collection and seating unit on which the concentration unit is rotatably seated; A collection guide part formed in the collection seating part and guiding the sample discharged from the concentration part; And a collection storage unit disposed below the collection and seating unit and storing the sample discharged from the concentration unit.

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

The storage unit is connected to the collection unit, and a storage cylinder having a storage space for storing at least one buffer; A storage passage unit 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 a 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 the buffer is introduced from the storage unit to purify the nucleic acid; And a purification discharging unit formed below the purification container and discharging the purified nucleic acid from the purification container.

The refining unit may further include a refining magnetic force unit that is detachable to the refining container and provides a magnetic force.

The 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 a 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 concentration unit to a purification unit; An input step of selectively inputting the buffer stored in the storage unit to the purification 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 gram-positive bacteria and gram-negative bacteria can be simultaneously concentrated in a sample by coating probes that bind to a specific layer on the surface of the pathogen.

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

The transfer step is characterized in that the magnetic force provided to the concentration unit is removed, the magnetic force is provided to the purification unit, and then the concentrated pathogen is transferred to the purification unit.

The nucleic acid extraction device and the nucleic acid extraction method according to the present invention are required for diagnostic tests because the low concentration pathogens present in a large volume sample can be rapidly separated from the sample by using magnetism and then concentrated to purify and separate the nucleic acid in the purification unit. You can save time and cost.

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 concentrations of pathogens are possible in a sample.

The nucleic acid extraction apparatus and the nucleic acid extraction method according to the present invention can store all buffers necessary for a storage unit integrally provided with the nucleic acid extraction apparatus, so that workability can be improved.

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

1 is a diagram schematically showing a nucleic acid extraction apparatus according to an embodiment of the present invention.
2 is a view schematically showing a concentration unit according to an embodiment of the present invention.
3 is a diagram schematically illustrating 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 purification unit according to an embodiment of the present invention.
6 is a flowchart schematically showing a nucleic acid extraction method according to an embodiment of the present invention.

Hereinafter, embodiments of a nucleic acid extracting apparatus and a nucleic acid extracting 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 components shown in the drawings may be exaggerated for clarity and convenience of description. In addition, terms to be described later are terms defined in consideration of functions in the present invention, which may vary according to the intention or custom of users or operators. Therefore, definitions of these terms should be made based on the contents throughout the present specification.

1 is a diagram schematically showing a nucleic acid extraction apparatus according to an embodiment of the present invention. Referring to FIG. 1, a nucleic acid extracting apparatus 1 according to an embodiment of the present invention includes a concentration unit 10, a collection 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 collection unit 20 on which the concentration unit 10 is rotatably mounted processes the sample discharged from the concentration 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, and purifies it with nucleic acid to discharge the purified nucleic acid.

The nucleic acid extracting device 1 having the above configuration can simultaneously multiplely concentrate pathogens of low concentration present in a sample (2-10 mL) to remove diagnostic inhibitors, and purify and extract nucleic acids from the concentrated pathogens. I can. Since the nucleic acid extracting apparatus 1 provides an integrated cartridge in which the above-described 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 device 1, the collection unit 20, the storage unit 30, and the purification unit 40 are integrally formed, and the concentration unit 10 is rotatably mounted on the collection unit 20. Can be. This nucleic acid extraction device 1 is produced by a 3D printer or injection molding, and when pathogens, which are bacteria, are concentrated in the concentration unit 10, the concentrated pathogens are transferred to the purification unit 40, and the washing process is performed. It proceeds with the purification and extraction process of the nucleic acid containing it.

At this time, nucleic acid means an acidic substance in the nucleus, and DNA and RNA are called nucleic acids. Nucleic acid is composed of a number of nucleotides linked together, and these nucleotides are the basic units that make up a nucleic acid and are composed of three parts: phosphoric acid, sugar, and base.

In addition, 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 organisms. Therefore, in order to analyze the composition, content, and properties of a gene, high-purity DNA must first be isolated. As the bacterial pathogen is dissolved, the nucleic acid is released, and the surface of the nucleic acid is negatively charged.

2 is a view schematically showing a concentration unit according to an embodiment of the present invention. Referring to Figure 2, the thickening unit 10 according to an embodiment of the present invention includes a thickening tube 11, a thickening insertion part 13, and a thickening outlet 14.

The concentrated container 11 stores samples containing magnetic nanoparticles and pathogens. For example, the thickening cylinder 11 may have a cylindrical shape and have an open upper side. The sample injected into the concentrated cylinder 11 uses magnetic nanoparticles coated with probes such as vancomycin and polymyxin that bind to a specific layer on the surface of the pathogen rather than a specific antigen. . Due to this, gram-positive bacteria and gram-negative bacteria may be simultaneously concentrated in the sample.

On the other hand, the thickening cover 12 may open and close the opened area of the thickening cylinder 11. For this reason, a sample including magnetic nanoparticles and pathogens can be injected into the open area through the condensing cover unit 12.

The concentration insertion portion 13 extends downward from the concentration cylinder portion 11, is inserted into the collection portion 20, and is rotatable in the collection portion 20. As an example, the concentration insertion portion 13 is smaller in diameter than the concentration cylinder portion 11 and the bottom portion of the concentration cylinder portion 11 may be seated on the collection portion 20.

The concentration outlet portion 14 is formed in the concentration insertion portion 13, and discharges a sample containing magnetic nanoparticles and pathogens. As an example, the concentration outlet 14 may be a hole or a pipe formed in the lower portion of the concentration insertion portion 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 to the condensing cylinder unit 11 and provides magnetic force. As an example, the magnetic enrichment unit 15 may be a permanent magnet or an electromagnet applied, and may be mounted on the outer circumferential surface of the enrichment cylinder 11. Pathogens connected to the magnetic nanoparticles by the magnetic enrichment unit 15 may be fixed to the inside of the enrichment cylinder 11 to be concentrated.

3 is a diagram schematically illustrating 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 and seating portion 21 is seated so that the concentration portion 10 is rotatable. As an example, the collection and seating portion 21 may be formed with a collection groove 211 so as to be rotatable by inserting the concentration insertion portion 13 in the central portion. Due to this, the edge of the thickening tube 11 may be seated at the edge of the collection and seating portion 21.

The collection guide part 22 is formed in the collection and seating part 21 and guides the sample discharged from the concentration part 10. As an example, the collection guide part 22 may be a hole or a pipe formed in the collection seating part 21, and the collection guide part 22 and the concentration outlet part 14 When communicated, samples excluding pathogens connected to the magnetic nanoparticles stored in the concentration tube part 11 and the concentration insertion part 13 may be discharged.

The collection and storage unit 23 is disposed below the collection and seating unit 21 and stores samples discharged from the concentration unit 10. As an example, the collection and storage unit 23 is detachable from the bottom of the collection and storage unit 21, and when a sample is stored in the collection and storage unit 23, the collection and storage unit 23 is separated from the collection and storage unit 21 After discarding the sample, the emptied collection and storage unit 23 may be remounted on the collection and seating 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 and 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 passage part 24 and the concentration outlet part 14 communicate with each other by the rotation of the concentration insertion part 13, the concentrated pathogen stored in the concentration cylinder part 11 and the concentration insertion 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, a storage unit 30 according to an embodiment of the present invention includes a storage cylinder 31, a storage passage 32, and a storage valve 33.

The storage cylinder 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 cylinder part 31 may include a storage shaft part 311 having a cylindrical shape, and a disk-shaped storage plate part 312 extending to an outer circumferential surface of the storage shaft part 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 passage unit 32 guides the buffer stored in the storage space 313 to the purification unit 40. For example, the storage passage unit 32 may have an upper end communicating with each storage space 313 and a lower end communicating with the storage shaft 311.

The storage valve unit 33 opens and closes the storage passage unit 32 to selectively discharge the buffer stored in the storage space 313. The storage valve unit 33 can be applied to a rotary valve method. As an example, the storage valve part 33 includes a valve shaft part 331 penetrating through the storage shaft part 311, a valve operation part 332 protruding laterally from the top of the valve shaft part 331 and operable, and a valve shaft part ( It may include a valve flow passage portion 333 formed in the 331 and selectively communicated with the storage passage portion 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 so that the buffer is converted into a purification part ( 40).

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

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

The purification discharge part 42 is formed under the purification container part 41 and discharges the purified nucleic acid from the purification container part 41. As an example, the tablet discharge unit 42 may include a discharge pipe part 421 connected to the lower portion of the tablet cylinder part 41 and a discharge valve part 422 that opens and closes the discharge pipe part 421.

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

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

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

When the sample preparation is complete, a sample supply step of supplying a sample including pathogens and magnetic nanoparticles to the concentration unit 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 condensing cylinder 11.

When a sample is supplied to the concentrating unit 10, the concentrating unit 10 has a concentration step of separating and concentrating pathogens and magnetic nanoparticles from the sample (S30). That is, 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 in the collection unit 20 and then discarded.

When the pathogen is concentrated through the concentration step, it has 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 concentration unit 10 is blocked, the magnetic force is provided to the purification unit 40, and then the concentrated pathogen is transferred to the purification unit 40.

That is, when the magnetic force provided to the concentrating unit 10 is blocked, the magnetic nanoparticles and pathogens concentrated in the concentrating unit 10 become movable. In addition, 1 mL of phosphate buffered saline (PBS) is used to move the concentrated pathogens and magnetic nanoparticles toward the purification unit 40. At this time, by rotating the concentration insertion portion 13, the concentration outlet portion 14 and the collection passage portion 24 are in communication,

PBS is a phosphate buffer, which is the most used buffer in biology-related 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 the addition of a certain amount of acid or base from the outside.

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

When the concentrated pathogen is transferred to the purification unit 40, the buffer stored in the storage unit 30 is selectively injected into the purification unit 40 (S50).

That is, various types of buffers are stored in the plurality of storage spaces 313 formed in the storage cylinder 31, and the buffers are selectively inputted to the purification unit 40 by the operation of the storage valve unit 33. I can.

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

Lysis buffer is a buffer used to destroy open cells for use in molecular biology experiments to 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 disintegrate the membrane structure, and this lysis buffer 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 cylinder 41 to bind to the nucleic acid.

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

Proteinase K is a proteolytic enzyme as a reagent to increase the purification efficiency of nucleic acids. Proteinase K is responsible for removing the histone protein attached to DNA. DNA has a histone protein attached to it to maintain stability. So, if Proteinase K is added, the purity drops when DNA is extracted.

RNase (ribonuclease) is an enzyme used to cut RNA. The reason why RNase is used for DNA purification is that when DNA is extracted, RNA is usually extracted using the same principle, and since DNA is needed, RNase is supplied to cut and remove unnecessary RNA, so pure DNA can be obtained.

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 is performed 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 then vortex is performed for 5 minutes.

Nucleic acid is extracted by dissolving the concentrated pathogen in this process, and the extracted nucleic acid binds to magnetic silica particles in an electrostatic bond form due to a cation bridge. Cationic crosslinking has a cation, and magnetic silica particles and nucleic acids have anions, so magnetic silica particles and nucleic acids are bound to both sides of the cation crosslinking.

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

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

Then, the magnetic force to the refining unit 40 is stopped and the vortex is performed for 5 minutes, and then the magnetic force is provided to the lower portion of the refining barrel 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 the washing buffer of 350 μL through the storage unit 30 is repeated three times, and all impurities in the purification cylinder 41 are discharged through the purification discharge unit 42. And the operation of removing the internal fluid in the refining cylinder part 41 is repeated three times.

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

In the nucleic acid extraction apparatus and nucleic acid extraction method according to an embodiment of the present invention, a low concentration pathogen present in a large-capacity sample is rapidly separated from the sample by using magnetism, and then concentrated to purify and separate the nucleic acid in the purification unit 40 Because it can be done, 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 surface of pathogens, multiple concentrations of pathogens in a sample are possible.

In addition, in the present invention, all buffers required for the storage unit 30 integrally provided with the nucleic acid extracting apparatus 1 can be stored, so that workability can be improved.

The present invention has been described with reference to the embodiments shown in the drawings, but these are only exemplary, and those of ordinary skill in the field to which the technology pertains, various modifications and other equivalent embodiments are possible. I will understand. Therefore, the true technical protection scope of the present invention should be determined by the following claims.

10: concentrated portion 11: concentrated barrel portion
12: thickening cover part 13: thickening insertion part
14: Concentration outlet 15: Concentration magnetic power section
20: collection unit 21: collection and seating unit
22: collection information unit 23: collection storage unit
24: collection flow section 30: storage section
31: storage container 32: storage passage
33: storage valve unit 40: purification unit
41: tablet container 42: tablet discharging part
43: refining magnetic field

Claims (13)

A concentration unit for storing a sample including magnetic nanoparticles and a pathogen, and for separating and concentrating the pathogen from the sample;
A collection unit mounted to be rotatable in the concentrating unit and processing the sample discharged from the concentrating unit;
A storage unit connected to the collection unit and capable of storing one or more buffers; And
And a purification unit connected to the storage unit and receiving the concentrated pathogen supplied from the concentration unit and purifying it with nucleic acid to discharge the purified nucleic acid.
The method of claim 1, wherein the concentration unit
A condensing tube unit in which a sample containing magnetic nanoparticles and pathogens is stored;
A concentration insertion portion extending downwardly from the concentration cylinder portion, inserted into the collection portion, and rotatable in the collection portion; And
A nucleic acid extraction apparatus comprising: a concentration outlet formed in the concentration insertion portion and for discharging a sample containing magnetic nanoparticles and pathogens.
The method of claim 2, wherein the concentration unit
Nucleic acid extraction apparatus further comprising a; detachable to the concentrated cylinder portion, the magnetic enrichment unit for providing a magnetic force.
The method of claim 1, wherein the collection unit
A collection and seating unit on which the concentration unit is rotatably seated;
A collection guide part formed in the collection seating part and guiding the sample discharged from the concentration part; And
And a collection storage unit disposed below the collection and seating unit and storing the sample discharged from the concentration unit.
The method of claim 4, wherein the collection unit
A nucleic acid extraction apparatus comprising: a collection passage formed in the collection and seating portion and guiding the concentrated pathogen discharged from the concentration portion to the purification portion.
The method of claim 1, wherein the storage unit
A storage cylinder connected to the collection unit and having a storage space for storing at least one buffer;
A storage passage unit 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 method of claim 6,
The storage valve unit, a nucleic acid extraction device, characterized in that the rotary valve method is applicable.
The method of claim 1, wherein the purification unit
A purification cylinder connected to a lower portion of the storage unit, through which pathogens concentrated in the concentration unit are introduced, and a buffer is introduced from the storage unit to purify nucleic acids; And
A nucleic acid extraction apparatus comprising: a purification discharge unit formed under the purification cylinder unit and for discharging the purified nucleic acid from the purification cylinder unit.
The method of claim 1, wherein the purification unit
A nucleic acid extracting apparatus further comprising a; a purification magnetic force portion detachable to the purification cylinder portion and providing a magnetic force.
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 a 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 concentration unit to a purification unit;
An input step of selectively inputting the buffer stored in the storage unit to the purification unit;
An extraction step of extracting nucleic acids from the pathogen transferred to the purification unit; And
A method for extracting nucleic acids comprising a; purification step of purifying the extracted nucleic acid.
The method of claim 10,
The magnetic nanoparticle is a nucleic acid extraction method, characterized in that the gram-positive bacteria and gram-negative bacteria can be simultaneously concentrated in the sample by coating probes that bind to a specific layer on the surface of the pathogen.
The method of claim 10, wherein the concentration step
Nucleic acid extraction method, characterized in that the magnetic nanoparticles are separated from the sample and concentrated by the magnetic force provided to the concentration unit.
The method of claim 10, wherein the transferring step
A nucleic acid extraction method comprising removing the magnetic force provided to the concentration unit, providing the magnetic force to the purification unit, and transferring the concentrated pathogen to the purification unit.

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