CN114276894A - Air pressure driven type rapid nucleic acid separation, purification, concentration and detection device - Google Patents

Air pressure driven type rapid nucleic acid separation, purification, concentration and detection device Download PDF

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Publication number
CN114276894A
CN114276894A CN202011034041.4A CN202011034041A CN114276894A CN 114276894 A CN114276894 A CN 114276894A CN 202011034041 A CN202011034041 A CN 202011034041A CN 114276894 A CN114276894 A CN 114276894A
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sample
pool
nucleic acid
washing liquid
pressurizing
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CN202011034041.4A
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王维鹏
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Individual
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Abstract

The invention provides a device which is not required to be used in a laboratory environment, can be used for driving a sample lysate, a washing solution and an eluent by air pressure, can be used for quickly and effectively removing interfering substances in the sample lysate, purifying and concentrating target nucleic acid, improving the speed and accuracy of nucleic acid detection and realizing the quick detection beside a pathogen nucleic acid bed or the household detection. Including shell body, sealing plug, sample pond, nucleic acid separation post, nucleic acid amplification pipe, side direction chromatography detection examination strip, nucleic acid detection drive liquid bag, slip cutter, eluant pond, washing liquid pond, supercharging device, sample pressurization connection structure, communicating pipe and check valve, in the shell body was arranged in to the sample pond, open at the shell body top has sample lysate to add the hole, sample lysate adds the hole and corresponds the intercommunication with sample pond top, and the nucleic acid separation post is fixed to be arranged in the sample pond, and is close to sample pond bottom, and in the shell body was arranged in to nucleic acid amplification pipe, and was located sample pond bottom under.

Description

Air pressure driven type rapid nucleic acid separation, purification, concentration and detection device
Technical Field
The invention relates to the field of nucleic acid detection in molecular biology, in particular to a device for rapidly separating, purifying, concentrating and directly detecting a specimen lysate by air pressure driving without a laboratory environment.
Background
The gold standard for detecting human infectious pathogens is culture and nucleic acid detection, and nucleic acid detection is the only gold standard for rapidly detecting infectious pathogens (such as viruses and the like). There is a need for point-of-care testing (POCT) or home testing of pathogens during epidemic infectious diseases due to medical resource limitations, or the disease involves personal privacy, or the disease requires continuous monitoring, among other reasons. Therefore, there is a need to develop a nucleic acid detection device that does not require a laboratory environment, has a shorter detection time, is more convenient to use, has a higher accuracy, and is safer for the iterative technique of nucleic acid detection. The difficulty in realizing the detection of POCT or domestic pathogen nucleic acid lies in the extraction and purification of the pathogen nucleic acid in the sample. Meanwhile, the concentration of the separated and purified sample nucleic acid is a key technology for improving the detection sensitivity.
With the development of science and technology, samples such as nasopharyngeal swab and various secretions can be sampled by a suspected patient or by a family for home assistance, and nucleic acid in the samples can be rapidly cracked and released, so that pathogenic microorganisms can be detected. However, the sample nucleic acid released by rapid lysis often contains various impurities that interfere with the sensitivity and specificity of detection, and needs to be purified and concentrated.
Therefore, the separation, purification and concentration of the sample nucleic acid after lysis and release are key technologies for improving the detection accuracy. Currently, this process can still only be accomplished in a laboratory environment. The method mainly comprises a magnetic bead method, a nucleic acid and nucleic acid separation column and a gel recovery method, wherein the magnetic bead method is difficult to miniaturize, and the gel recovery method is only generally used for scientific research in a laboratory environment.
All the current sample nucleic acid purification and concentration processes have the following disadvantages: 1. needs to be done in a laboratory environment to avoid cross contamination and biosafety risks; 2. a variety of instrumentation is required; 3. a professional technician is required to perform operations of a plurality of steps; 4. requiring a long time to queue up for specimen collection or delivery; 5. it takes a long time (at least 3-4 hours) to wait for the detection result. Therefore, a device for rapidly purifying and concentrating nucleic acid from a sample lysate, which is simple, fast, safe and effective without a laboratory environment, is needed.
Disclosure of Invention
In view of the technical deficiencies, the present invention provides an air pressure driven type rapid nucleic acid separation, purification, concentration and detection device, which is not required to be used in a laboratory environment, and can rapidly and effectively remove interfering substances in a sample lysate, purify and concentrate target nucleic acid by driving the sample lysate, a washing solution and an eluent through air pressure, thereby increasing the nucleic acid detection speed and accuracy, and realizing rapid detection beside a pathogen nucleic acid bed or home detection.
In order to solve the technical problems, the invention adopts the following technical scheme:
a pneumatic-driven type rapid nucleic acid separation, purification, concentration and detection device is characterized by comprising a shell, a sealing plug, a sample pool, a nucleic acid separation column, a nucleic acid amplification tube, a lateral chromatography detection test strip, a nucleic acid detection driving liquid bag, a sliding cutter, an eluent pool, a washing liquid pool, a pressurizing device, a sample pressurizing and communicating structure, a communicating tube and a one-way valve, wherein the sample pool is arranged in the shell, the top of the shell is provided with a sample lysate adding hole, the sample lysate adding hole is correspondingly communicated with the top end of the sample pool, the nucleic acid separation column is fixedly arranged in the sample pool and is close to the bottom end of the sample pool, the nucleic acid amplification tube is arranged in the shell and is positioned right below the bottom end of the sample pool, the lateral chromatography detection test strip is arranged on the bottom surface of the inner side of the shell and corresponds to the nucleic acid amplification tube, the nucleic acid detection driving liquid bag is arranged in the shell and is adjacent to the nucleic acid amplification tube, the nucleic acid detection driving liquid bag is positioned above the head end of the lateral chromatography detection test strip, the sliding cutter is arranged in the outer shell and positioned below the nucleic acid detection driving liquid bag, the lateral chromatography detection test strip is provided with a detection line and a contrast line, the sealing plug penetrates through the sample lysate adding hole and is arranged at the top end of the sample pool, the outer side surface of the sealing plug is provided with a sealing plug clamping block, the position on the sample pool corresponding to the sealing plug clamping block is provided with a sealing plug bayonet, the sealing plug clamping block is clamped in the sealing plug bayonet, the eluent pool and the washing liquid pool are arranged in the outer shell and are respectively communicated with the sample pool through a communication pipe, the communication positions of the sample pool and the communication pipe are provided with one-way valves, the sample pressurization communication structure is communicated with the sample pool, the communication positions are provided with the one-way valves, and the pressurization device is communicated with the sample pressurization communication structure, the eluent pool and the washing liquid pool, the eluent pool is internally provided with eluent, the washing liquid pool is internally provided with washing liquid, and the washing liquid pool is composed of a washing liquid A pool and a washing liquid B pool;
preferably, the pressurizing device is a built-in pressurizing air bag which is positioned in the outer shell;
preferably, three built-in pressurizing air bags are arranged in the outer shell corresponding to the eluent pool, the washing liquid pool and the sample pressurizing and communicating structure, the washing liquid A pool and the washing liquid B pool are communicated with one built-in pressurizing air bag through a three-way pipe, and an air inlet pipe is arranged on each built-in pressurizing air bag and extends to penetrate through the wall of the sample pool;
preferably, an inner pressurizing air bag is arranged in the outer shell corresponding to the washing liquid A pool, the washing liquid B pool and the eluent pool, and an air inlet pipe is arranged on the inner pressurizing air bag and extends to penetrate through the wall of the sample pool;
preferably, the pressurizing device is an external pressurizing air bag, the external pressurizing air bag is positioned outside the outer shell, an inflation tube is arranged on the external pressurizing air bag and is communicated with the specimen pressurizing and communicating structure, the washing liquid A pool, the washing liquid B pool and the eluent pool;
preferably, the specimen pressurizing communicating structure is a specimen pressurizing pipeline;
preferably, the specimen pressurizing and communicating structure is a one-way through groove, the one-way through groove is arranged in the middle of the sealing plug and longitudinally penetrates through the sealing plug, and a one-way valve is arranged at the bottom end of the one-way through groove;
preferably, the bottom end of the specimen pool is of a conical structure;
preferably, the specimen pool consists of an upper part and a lower part, the diameter of the upper part of the specimen pool is larger than that of the lower part of the specimen pool, and the bottom end of the upper part of the specimen pool is of a conical structure;
preferably, the diameter of the nucleic acid amplification tube is gradually reduced from top to bottom.
The invention has the beneficial effects that: 1. the method is not required to be carried out in a laboratory environment, and the rapid detection beside a pathogen nucleic acid bed or the home detection is realized.
2. The use is simple and quick, safe and effective, and professional training is not needed.
3. The sample lysate, the washing solution and the eluent are driven by air pressure, so that interfering substances in the sample lysate can be quickly and effectively removed, the target nucleic acid is purified and concentrated, and the nucleic acid detection speed and accuracy are improved.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below, it is obvious that the drawings in the following description are only some embodiments of the present invention, and for those skilled in the art, other drawings can be obtained according to the drawings without creative efforts.
FIG. 1 is a schematic structural diagram of a pneumatic-driven rapid nucleic acid separation, purification, concentration and detection device provided in embodiment 1 of the present invention;
FIG. 2 is a schematic structural diagram of a pneumatic-driven apparatus for rapid separation, purification, concentration and detection of nucleic acid according to embodiment 2 of the present invention;
FIG. 3 is a schematic structural diagram of a pneumatic-driven apparatus for rapid separation, purification, concentration and detection of nucleic acid according to embodiment 3 of the present invention;
description of reference numerals:
the device comprises an outer shell 1, a sealing plug 2, a sealing plug bayonet 3, a sample cell 4, a nucleic acid separation column 5, a detection line 6, a comparison line 7, a nucleic acid amplification tube 8, a lateral chromatography detection test strip 9, a nucleic acid detection driving liquid bag 10, a sliding cutter 11, an eluent cell 12, a washing liquid B cell 13, a washing liquid A cell 14, an external pressurizing air bag 15, a communicating tube 16, a sample pressurizing pipeline 17, a one-way valve 18, a one-way through groove 19, a sample lysate adding hole 20, an internal pressurizing air bag 21, a three-way pipe 22 and an air inlet pipe 23.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
Example 1
As shown in FIG. 1, a pneumatic-driven rapid nucleic acid separation, purification, concentration and detection device comprises an outer shell 11, a sealing plug 22, a sample cell 44, a nucleic acid separation column 55, a nucleic acid amplification tube 88, a lateral chromatography detection strip 99, a nucleic acid detection driving liquid bag 1010, a sliding cutter 1111, an eluent cell 1212, a washing liquid cell, a pressurizing device, a sample pressurizing and communicating structure, a communicating tube 1616 and a one-way valve 1818, wherein the sample cell 4 is arranged in the outer shell 1, the top of the outer shell 1 is provided with a sample lysate adding hole 20, the sample lysate adding hole 20 is correspondingly communicated with the top end of the sample cell 4, the nucleic acid separation column 5 is fixedly arranged in the sample cell 4 and close to the bottom end of the sample cell 4, a nucleic acid amplification tube 8 is arranged in the outer shell 1 and is positioned right below the bottom end of the sample cell 4, the lateral chromatography detection strip 9 is arranged on the bottom surface of the inner side of the outer shell 1 and corresponds to the nucleic acid amplification tube 8, the nucleic acid detection driving liquid bag 10 is arranged in the outer shell 1 and is adjacent to the nucleic acid amplification tube 8, the nucleic acid detection driving liquid bag 10 is positioned above the head end of a lateral chromatography detection test strip 9, the sliding cutter 11 is arranged in the outer shell 1 and is positioned below the nucleic acid detection driving liquid bag 10, a detection line 6 and a contrast line 7 are arranged on the lateral chromatography detection test strip 9, the sealing plug 2 penetrates through a sample lysate adding hole 20 and is arranged at the top end of the sample cell 4, a sealing plug 2 clamping block is arranged on the outer side surface of the sealing plug 2, a sealing plug bayonet 3 is arranged on the sample cell 4 corresponding to the sealing plug 2 clamping block, the sealing plug 2 clamping block is arranged in the bayonet 3, the eluent cell 12 and the washing liquid cell are arranged in the outer shell 1 and are respectively communicated with the sample cell 4 through a communication tube 16, and a one-way valve 18 is arranged at the communication position of the sample cell 4 and the communication tube 16, sample pressurization extending structure and sample pond 4 are linked together, and the intercommunication position has put check valve 18, supercharging device and sample pressurization extending structure, eluant pond 12, washing liquid pond are linked together, eluant pond 12 embeds there is the eluant, the washing liquid pond embeds there is the washing liquid, the washing liquid pond comprises washing liquid A pond 14 and washing liquid B pond 13, the supercharging device is external pressure boost gasbag 15, external pressure boost gasbag 15 is located the shell body 1 outside, be provided with an gas tube on the external pressure boost gasbag 15, and all communicate with sample pressurization extending structure, washing liquid A pond 14, washing liquid B pond 13, elution liquid pond 12, sample pressurization extending structure is sample pressurization pipeline 17.
When in use, firstly, the collected specimen, such as a nasopharyngeal swab, a pharyngeal swab and the like, is put into a specimen lysate (usually 1-3 ml), and is properly stirred and uniformly mixed; then sucking the sample lysate (about 1-2 ml) by using a plastic straw, and adding the sample lysate into the sample pool 4 through a sample lysate adding hole 20; then pushing the sealing plug 2 into the opening 4 of the specimen pool and clamping the sealing plug 2 by the clamping block 3;
firstly, pressing a pressurizing device to pressurize the sample tank 4 by gas through a sample pressurizing and communicating structure, driving the sample lysate to pass through a nucleic acid separation column 5, leaving nucleic acid and partial impurities in the sample on the nucleic acid separation column 5, and enabling the waste liquid to flow out of the sample tank 4 to enter a waste liquid tank;
then the washing liquid in the cavity A of the washing liquid and the cavity B of the washing liquid are sequentially driven by the supercharging device to enter the sample tank 4, the washing liquid passes through the nucleic acid separation column 5 to take away impurities in the nucleic acid separation column 5, and the washing liquid flows out of the sample tank 4 to enter the waste liquid tank; the pressurizing structure can be repeatedly pressed for multiple times, so that the washing liquid completely flows through the nucleic acid separation column 5, and the residual washing liquid is fully volatilized; and the washing steps can be repeated or increased or the types of washing solutions can be increased according to the components of the lysis solution used by the specimen lysis solution or the standard of the nucleic acid purity required to be obtained;
finally, aligning the sample pool 4 with the nucleic acid amplification tube 8, pressing a pressurizing device to drive eluent (about 30-100 microliters) to pass through the nucleic acid separation column 5, dissolving and eluting nucleic acid in the nucleic acid separation column 5 to obtain about 30-100 microliters of purified and concentrated nucleic acid solution, and flowing the purified and concentrated nucleic acid solution into the nucleic acid amplification tube 8;
after the amplification is finished, the sliding cutter 11 is pushed to cut open the nucleic acid detection driving liquid bag 10 and the amplification tube base, the nucleic acid amplification product flows to the lateral chromatography detection test strip 9, the driving liquid flows to the lateral chromatography detection test strip 9, the motion of the amplified product liquid on the lateral chromatography detection test strip 9 is pushed, the driving liquid drives the nucleic acid amplification product to move on the lateral chromatography detection test strip, and therefore the result display is achieved.
The shape of the device can be designed according to the needs, and can be irregular, elliptical, square and the like;
example 2
The difference between this example and example 1 is: the pressurizing device is a built-in pressurizing air bag 21, the built-in pressurizing air bag 21 is positioned in the outer shell 1, an inner pressurizing air bag is arranged in the outer shell 1 corresponding to the washing liquid A pool 14, the washing liquid B pool 13 and the eluent pool 12, an air inlet pipe 23 is arranged on the inner pressurizing air bag and extends to penetrate through the wall of the specimen pool 4, the specimen pressurizing and communicating structure is a one-way through groove 19, the one-way through groove 19 is arranged in the middle of the sealing plug 2 and longitudinally penetrates through the sealing plug 2, and a one-way valve 18 is arranged at the bottom end of the one-way through groove 19; when in use, the washing liquid and the eluent are pushed one by one through the inner pressurizing air bags by the pressurizing devices such as the air pipes, and the pressurizing devices blow air to the one-way through groove 19 to pressurize the specimen pool 4;
example 3
The difference between this example and example 2 is: the washing liquid A pool 14 and the washing liquid B pool 13 are communicated with the corresponding built-in pressurizing air bag 21 through a three-way pipe 22, and an air inlet pipe 23 is arranged on the built-in pressurizing air bag 21 and extends to penetrate through the wall of the sample pool 4; when in use, the three-way pipe 22 is rotated to change the communicated washing liquid A pool 14 and washing liquid B pool 13, so that the number of the check valves 18 is reduced;
further, the bottom end of the specimen pool 4 is of a conical structure;
further, the specimen pool 4 consists of an upper part and a lower part, the diameter of the upper part of the specimen pool 4 is larger than that of the lower part, and the bottom end of the upper part of the specimen pool 4 is of a conical structure;
further, the diameter of the nucleic acid amplification tube 8 is gradually reduced from the top to the bottom.
The design operator can operate without professional technicians or special training by referring to a simple picture instruction sheet, for example, 1-2 keys can be continuously pressed according to the instruction sheet, and the processes of nucleic acid separation, purification, concentration and detection can be completed in a totally closed environment.
It will be apparent to those skilled in the art that various changes and modifications may be made in the present invention without departing from the spirit and scope of the invention. Thus, if such modifications and variations of the present invention fall within the scope of the claims of the present invention and their equivalents, the present invention is also intended to include such modifications and variations.

Claims (10)

1. A pneumatic drive type rapid nucleic acid separation, purification, concentration and detection device is characterized by comprising a shell, a sealing plug, a sample pool, a nucleic acid separation column, a nucleic acid amplification tube, a lateral chromatography detection test strip, a nucleic acid detection driving liquid bag, a sliding cutter, an eluent pool, a washing liquid pool, a pressurizing device, a sample pressurizing and communicating structure, a communicating tube and a one-way valve, wherein the sample pool is arranged in the shell, the top of the shell is provided with a sample lysate adding hole, the sample lysate adding hole is correspondingly communicated with the top end of the sample pool, the nucleic acid separation column is fixedly arranged in the sample pool and is close to the bottom end of the sample pool, the nucleic acid amplification tube is arranged in the shell and is positioned right below the bottom end of the sample pool, the lateral chromatography detection test strip is arranged on the bottom surface of the inner side of the shell and corresponds to the nucleic acid amplification tube, and the nucleic acid detection driving liquid bag is arranged in the shell, the lateral chromatography detection test strip is provided with a detection line and a contrast line, a sealing plug penetrates through a sample lysate adding hole and is arranged at the top end of the sample pool, the outer side surface of the sealing plug is provided with a sealing plug clamping block, the position on the sample pool corresponding to the sealing plug clamping block is provided with a sealing plug bayonet, the sealing plug clamping block is clamped in the sealing plug bayonet, the eluent pool and the washing liquid pool are arranged in the outer shell and are respectively communicated with the sample pool through a communication pipe, the communication position of the sample pool and the communication pipe is provided with a one-way valve, the sample pressurization communication structure is communicated with the sample pool, the communication position is provided with a one-way valve, and the pressurization device and the sample pressurization communication structure, The elution liquid pool is communicated with the washing liquid pool, the elution liquid is placed in the elution liquid pool, the washing liquid pool is placed with the washing liquid, and the washing liquid pool is composed of a washing liquid pool A and a washing liquid pool B.
2. The apparatus according to claim 1, wherein the pressurizing means is a built-in pressurizing air bag, and the built-in pressurizing air bag is located in the outer housing.
3. The apparatus according to claim 2, wherein three internal pressurizing air bags are provided in the outer housing corresponding to the eluent pool, the washing liquid pool and the sample pressurizing and communicating structure, the washing liquid A pool and the washing liquid B pool are communicated with a corresponding one of the internal pressurizing air bags through a three-way pipe, and an air inlet pipe is provided on the internal pressurizing air bag and extends to penetrate through the wall of the sample pool.
4. The apparatus according to claim 2, wherein an inner pressurizing air bag is disposed in the outer casing corresponding to the washing liquid A pool, the washing liquid B pool, and the elution liquid pool, and an air inlet tube is disposed on the inner pressurizing air bag and extends to penetrate the wall of the sample pool.
5. The apparatus according to claim 3 or 4, wherein the sample pressurizing and communicating structure is a sample pressurizing tube.
6. The apparatus according to claim 1, wherein the pressurizing device is an external pressurizing air bag, the external pressurizing air bag is located outside the outer casing, and an air tube is disposed on the external pressurizing air bag and is in communication with the specimen pressurizing and communicating structure, the washing liquid A pool, the washing liquid B pool, and the elution liquid pool.
7. The apparatus according to claim 6, wherein the sample pressurizing and communicating structure is a one-way channel, the one-way channel is disposed in the middle of the sealing plug and longitudinally penetrates the sealing plug, and a one-way valve is disposed at the bottom of the one-way channel.
8. The apparatus according to claim 1, wherein the bottom of the sample cell is of a cone-shaped structure.
9. The apparatus according to claim 1 or 8, wherein the sample cell is composed of an upper portion and a lower portion, the diameter of the upper portion of the sample cell is larger than that of the lower portion, and the bottom end of the upper portion of the sample cell is a cone-shaped structure.
10. The apparatus of claim 1, wherein the diameter of the nucleic acid amplification tube decreases from top to bottom.
CN202011034041.4A 2020-09-27 2020-09-27 Air pressure driven type rapid nucleic acid separation, purification, concentration and detection device Pending CN114276894A (en)

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CN202011034041.4A CN114276894A (en) 2020-09-27 2020-09-27 Air pressure driven type rapid nucleic acid separation, purification, concentration and detection device

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Application Number Priority Date Filing Date Title
CN202011034041.4A CN114276894A (en) 2020-09-27 2020-09-27 Air pressure driven type rapid nucleic acid separation, purification, concentration and detection device

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CN114276894A true CN114276894A (en) 2022-04-05

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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN114653093A (en) * 2022-04-19 2022-06-24 河南农业大学 Extraction and purification device of polysaccharide

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN114653093A (en) * 2022-04-19 2022-06-24 河南农业大学 Extraction and purification device of polysaccharide
CN114653093B (en) * 2022-04-19 2023-02-28 河南农业大学 Extraction and purification device of polysaccharide

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