CN114437916A

CN114437916A - Flow direction control system for nucleic acid extraction device

Info

Publication number: CN114437916A
Application number: CN202110276843.4A
Authority: CN
Inventors: 张祥林; 黄华生; 张一桐; 黄天茹
Original assignee: Individual
Current assignee: Individual
Priority date: 2020-11-03
Filing date: 2021-03-15
Publication date: 2022-05-06
Also published as: CN217418702U; CN114437901A; CN216947041U; CN114437900A; CN217418701U

Abstract

The invention relates to the field of nucleic acid extraction devices, in particular to a flow direction control system for a nucleic acid extraction device, which comprises a reaction bin main body, a recovery bin main body and a waste liquid pool main body, wherein the reaction bin main body is communicated with the recovery bin main body through a second flow channel main body, the reaction bin main body is communicated with the waste liquid pool main body through a first flow channel main body, a first air pressure regulation reserved part is arranged on the recovery bin main body, and a second air pressure regulation reserved part is arranged on the waste liquid pool main body. The flow direction control system for the nucleic acid extraction device ensures that the waste liquid does not remain in the reaction bin and flows into the waste liquid pool completely; can avoid the backward flow of reagent to the at utmost, can also be in when retrieving the export in storehouse and the reaction storehouse of waste liquid pond sharing, avoid the waste liquid in waste liquid pond to get into and retrieve the storehouse. The flow direction control system for a nucleic acid isolation apparatus of the present invention can also be applied to a nucleic acid isolation apparatus.

Description

Flow direction control system for nucleic acid extraction device

Technical Field

The invention relates to the field of nucleic acid extraction devices, in particular to a flow direction control system for a nucleic acid extraction device.

Background

The nucleic acid extraction is an important application circle in life science research, gene detection and the like, and the early nucleic acid extraction mainly removes impurities through phenol and chloroform extraction and precipitates nucleic acid through ethanol, but the method is complex to operate. At present, nucleic acid extraction reagents in the market mainly comprise nucleic acid extraction based on an adsorption column method and a magnetic bead method, and 1) the adsorption column method: the method has high extraction efficiency, is widely used at present, but needs repeated centrifugation, and is not beneficial to high-flux and automatic operation; 2) magnetic bead method: the method is based on magnetic adsorption, is easy to realize automatic and high-flux operation, but has higher cost and easily influenced extraction efficiency. In addition, the two methods are complex and tedious to operate, have certain technical operation difficulty, need to match professional technicians and many matched instruments and equipment, are time-consuming and labor-consuming in extraction, have high requirements on laboratory environment (such as cross contamination is easy to generate in the extraction process), especially have the requirement of equipping a special PCR laboratory (requiring a special sample preparation room or even requiring a negative pressure environment) for the clinical infectious sample for clinical examination, are difficult to adapt to the detection, timeliness and biological safety controllability requirements of clinical sample types with diversity, are difficult to deal with emergencies, and are not favorable for the realization of instant nucleic acid detection (POCT). The problems of waste liquid discharge and nucleic acid recovery during nucleic acid extraction have great influence on the real-time detection of nucleic acid, and the flow direction control system for a simple and controllable nucleic acid extraction device cannot be found in the prior art to solve the problems.

Disclosure of Invention

In view of the above-described drawbacks of the prior art, it is an object of the present invention to provide a flow direction control system for a nucleic acid extracting apparatus, which solves the problems of the prior art.

In order to achieve the above and other related objects, the present invention provides a flow direction control system for a nucleic acid extraction device, the flow direction control system for a nucleic acid extraction device includes a reaction bin main body, a recovery bin main body, and a waste liquid pool main body, the reaction bin main body is communicated with the recovery bin main body through a second flow channel main body, the reaction bin main body is communicated with the waste liquid pool main body through a first flow channel, a first air pressure regulation reserved portion is provided on the recovery bin main body, and a second air pressure regulation reserved portion is provided on the waste liquid pool main body.

As described above, the flow direction control system for a nucleic acid isolation apparatus according to the present invention has the following advantageous effects: ensuring that the waste liquid does not remain in the reaction bin and flows into a waste liquid pool completely; can avoid the backward flow of reagent to the at utmost, can also be in when retrieving the export in storehouse and the reaction storehouse of waste liquid pond sharing, avoid the waste liquid in waste liquid pond to get into and retrieve the storehouse. The method can be applied to a nucleic acid extraction device, and the extracted and recovered nucleic acid can be applied to downstream PCR amplification of different types, rapid detection of pathogenic microorganisms and the like, and has high integration level and easy use. The flow direction control system for the nucleic acid isolation apparatus of the present invention also has a certain revelation effect on the control of the flow direction of a liquid in other fields.

Drawings

FIG. 1 is a front view of a flow direction control system for a nucleic acid isolation apparatus according to the present invention.

FIG. 2 is a front view of the nucleic acid extracting apparatus of the present invention.

FIG. 3 is a bottom view of the nucleic acid isolation apparatus of the present invention.

FIG. 4 is a side view of the nucleic acid isolation apparatus of the present invention.

Description of the element reference numerals

1 reagent storehouse body

2 reaction cabin body

21 reaction chamber pressure regulating part

3 recovery bin main body

31 first air pressure adjusting reserve part

4 waste liquid pool main body

41 second air pressure regulating reserve part

5 support plate

6 first flow passage body

7 second flow passage body

71 reaction bin output section

72 upwarping section

73 recycling bin input section

Included angle between output section and upwarp section of alpha reaction bin

Beta the included angle between the upwarping section and the input section of the recovery bin

Included angle between gamma first flow channel main body and reaction bin output section

Detailed Description

The following description of the embodiments of the present invention is provided for illustrative purposes, and other advantages and effects of the present invention will become apparent to those skilled in the art from the present disclosure.

Please refer to fig. 1 to 4. It should be understood that the structures, ratios, sizes, and the like shown in the drawings and described in the specification are only used for matching with the disclosure of the specification, so as to be understood and read by those skilled in the art, and are not used to limit the conditions under which the present invention can be implemented, so that the present invention has no technical significance, and any structural modification, ratio relationship change, or size adjustment should still fall within the scope of the present invention without affecting the efficacy and the achievable purpose of the present invention. In addition, the terms "upper", "lower", "left", "right", "middle" and "one" used in the present specification are for clarity of description, and are not intended to limit the scope of the present invention, and the relative relationship between the terms and the terms is not to be construed as a scope of the present invention.

As shown in fig. 1, the present invention provides a flow direction control system for a nucleic acid extraction device, the flow direction control system for a nucleic acid extraction device includes a reaction chamber main body 2, a recovery chamber main body 3, and a waste liquid pool main body 4, the reaction chamber main body 2 is communicated with the recovery chamber main body 3 through a second flow channel main body 7, the reaction chamber main body 2 is communicated with the waste liquid pool main body 4 through a first flow channel main body 6, a reaction chamber air pressure adjusting portion 21 is provided on the reaction chamber main body 2, a first air pressure adjusting reserved portion 31 is provided on the recovery chamber main body 3, and a second air pressure adjusting reserved portion 41 is provided on the waste liquid pool main body 4.

Nucleic acid extraction element includes backup pad 5, be formed with reagent storehouse main part 1, reaction storehouse main part 2, recovery storehouse main part 3 and waste liquid pool main part 4 in the backup pad 5, be equipped with the runner of intercommunication between reagent storehouse main part 1 and the reaction storehouse main part 2, reaction storehouse main part 2 all communicates with recovery storehouse main part 3, waste liquid pool main part 4.

In one embodiment, a sealing film is provided on the support plate 5. The sealing film is pasted on the supporting plate 5 and is matched with the supporting plate 5 to form a complete reagent bin, a reaction bin, a recovery bin and a waste liquid pool. The sealing film is bonded and sealed with the support plate 5 by means of laser, asymmetric heat sealing or ultrasonic welding and the like. The material of the sealing film is selected from plastic, aluminum foil or composite materials of the plastic and the aluminum foil. The sealing film is not broken when the reagent cartridge main body 1 is extruded and deformed.

In one embodiment, the support plate, the reaction chamber body, the recovery chamber body and the waste liquid tank body may be formed separately and then bonded together by laser, thermal or sonic welding, etc. In another embodiment, the support plate 5 is integrally formed with the reagent cartridge body 1, the reaction cartridge body 2, the recovery cartridge body 3, and the waste liquid tank body 4. A reagent bin main body 1, a reaction bin main body 2, a recovery bin main body 3 and a waste liquid pool main body 4 are formed on the supporting plate 5. In one embodiment, the reagent bin main body 1, the reaction bin main body 2, the recovery bin main body 3 and the waste liquid pool main body 4 are all open bin bodies formed by a supporting plate 5 through a plastic suction or blow molding process and protruding out of the supporting plate 5, and after the supporting plate 5 is covered by a sealing film, the reagent bin main body 1, the reaction bin main body 2, the recovery bin main body 3 and the waste liquid pool main body 4 form a closed bin body. The reagent cabin main body 1, the reaction cabin main body 2, the recovery cabin main body 3, the waste liquid pool main body 4, the first flow channel main body 6 and the second flow channel main body 7 are all protruded from the supporting plate 5 on one side.

The material of the supporting plate 5 is selected from flexible materials. The material is selected to be effective in preventing volatilization of the reagent and to be non-reactive with the pre-loaded reagent. In one embodiment selected from flexible plastic films. Such as polyester, polyethylene terephthalate (PET), polycarbonate, polypropylene, polymethyl methacrylate, and mixtures thereof. In another embodiment, the material of the supporting plate 5 is selected from aluminum films. The supporting plate 5 can be formed by bonding two or more layers of materials, so that each chamber and flow channel formed by the supporting plate can be used as a carrier for liquid mixing, transferring and reacting.

The wall thickness of the supporting plate 5 except for the bin bodies is 0.8-2.0 mm.

The reaction bin air pressure adjusting part 21 is arranged at the top of the reaction bin. The arrangement position of the reaction chamber air pressure adjusting part 21 ensures that the substance added into the reaction chamber does not flow out. The number of the reaction chamber air pressure regulating parts 21 is one or more. In one embodiment, there is one reaction chamber pressure adjusting unit 21. The reaction chamber air pressure adjusting part 21 is used for air intake and adding a sample of nucleic acid to be extracted. The reaction chamber air pressure adjusting part 21 can be used in cooperation with a pressurizing member. The pressurizing component can blow and pressurize the air pressure adjusting part 21 of the reaction bin after the magnetic bead cleaning is finished so as to air-dry the magnetic beads and the residual cleaning solution; the reaction chamber pressure adjusting section 21 may be inflated and pressurized during nucleic acid recovery to allow the product to flow into the recovery chamber. In one embodiment, the pressurizing member is selected from a pressurizing bladder or air pump that can provide pneumatic pressure. The pressing member includes a pressing member head and a pressing member body. The pressing member body may be manufactured by any means known in the art, such as, but not limited to, injection molding, die casting, 3D printing, or machining. The compression member body may be made of any durable material known in the art, such as, but not limited to, plastics (e.g., polyethylene, polypropylene, polystyrene, polycarbonate, ABS, PEEK, etc.), metals (e.g., aluminum alloys, copper-lead alloys, etc.), and metal and plastic composites. The head of the pressurizing member can be made of elastic materials, so that the head of the pressurizing member is consistent with the air pressure adjusting part of the reaction bin in a sealing fit manner when air pressure is applied, and air pressure leakage is avoided. Alternative resilient materials that may be used for the head of the compression member include, but are not limited to, neoprene, polyurethane foam, polyimide foam, foam rubber, and the like.

Magnetic beads are added to the reaction chamber during nucleic acid extraction for recovery and purification of nucleic acids from the lysate. The use of magnetic beads requires the nucleic acid extraction device to be fitted with a suitable magnet drive system. The magnet driving system is, for example, a strong permanent magnet device, an electromagnet or a magnet array composed of permanent magnet and electromagnet. Preferably, the magnet driving system is an electromagnet. Can realize being higher than the adjustable magnetic force of permanent magnetism through increaseing the electric current, can realize 95% nanometer magnetic bead absorption in 30 seconds, consequently be favorable to improving the magnetic adsorption efficiency in the nucleic acid extraction, avoid magnetic force to attenuate along with the extension of distance fast.

In one embodiment, the reaction chamber has a flat profile rather than a profile with a curved surface. Namely, all the surfaces of the reaction bin are planes. The flat design enables the reaction chamber to be used with a magnet drive system. In one embodiment, the wall thickness of the reaction bin is 0.5-1.5 mm.

The flat design of reaction storehouse can also make the reaction storehouse cooperate the high frequency oscillation device to use, and accessible high frequency vibration reduces the wall built-up of liquid, avoids liquid to concentrate on the bottom of bag chamber under the action of gravity when, because the liquid bead wall built-up phenomenon that appears in the existence of surface tension, reduces the error, improves the rate of recovery of nucleic acid.

In one embodiment, the reaction chamber is prismatic in shape. The preferred shape is a triangular prism. The shape of triangular prism is favorable to the liquid flow in reagent storehouse to react the storehouse, thereby is favorable to also reacting the liquid gathering in the storehouse and discharging the liquid in the storehouse more fast to an arris.

The reaction bin can also be used with a heater. The heater is used for controlling the reaction temperature in the reaction bin. The heater may be a peltier device, a resistive heater, an electromagnetic heater, a thin film heater, and a positive temperature coefficient heater or other heater known in the art.

The bottom of the reaction bin is also provided with an outlet 22. And the outlet 22 is provided at the distal end of the reaction chamber air pressure adjusting portion 21. The liquid in the reaction bin can not flow out randomly. The outlet 22 is in sealed connection with the second flow passage.

The reaction bin main body is communicated with the recovery bin main body through a second flow passage main body 7.

In one embodiment, the second flow channel main body 7 comprises a reaction bin output section 71, an upturned section 72 and a recovery bin input section 73 which are sequentially communicated. The included angle alpha between the reaction bin output section 71 and the upwarping section 72 is an acute angle. The included angle beta between the upwarp section 72 and the recovery bin input section 73 is an acute angle.

The design of acute angle can avoid the backward flow of reagent in the at utmost between 7 sections of second runner main part, can also avoid when retrieving the export in storehouse and the reaction storehouse of waste liquid pond sharing, the waste liquid in waste liquid pond gets into retrieves the storehouse.

The recovery bin main body 3 is provided with a first air pressure adjusting reserved part 31. The machine can pierce through the first air pressure regulation reserve part 31 to make the gas enter the recovery bin, and the pressure in the recovery bin is balanced with the atmospheric pressure. Before the nucleic acid extraction device is not used, the first air pressure regulation reservation part 31 is in a closed state, and liquid does not flow to the recovery bin at the moment because the interior of the recovery bin is not balanced with the atmospheric pressure. The first pneumatic adjustment reserve 31 may be pierced to be opened according to experimental needs. For example, when the nucleic acid extraction is completed and the extracted nucleic acid is to be recovered, the first air pressure regulating reserve part 31 is opened to guide the nucleic acid to the recovery chamber. The first air pressure adjustment allowance part 31 may also be used in cooperation with a pressing member. The first air pressure adjusting reservation part 31 may be a reservation hole on the recovery bin, or may be a branch pipe protruding from the recovery bin as shown in fig. 1.

The waste liquid pool main body is communicated with the reaction bin main body through a first flow passage main body. The first flow passage main body is a linear flow passage. An included angle gamma between the first flow channel main body and the reaction bin output section 71 is an obtuse angle. Namely, the first flow channel main body is of a sinking type design. When the flow direction control system for the nucleic acid extraction device is in a use state, the first flow channel and the waste liquid pool are arranged below the reaction bin, so that no waste liquid such as cleaning liquid of magnetic beads and the like is left in the reaction bin.

And a second air pressure adjusting reserved part 41 is arranged on the waste liquid tank main body 4. The second air pressure adjustment reservation part 41 may be opened or closed as needed. When the reaction bin needs to discharge the waste liquid, the second air pressure adjusting reserved part 41 is opened, the air pressure in the waste liquid pool 4 is balanced with the atmospheric pressure, the second flow channel main body 7 is provided with the upwarping section 72, and at the moment, the first air pressure adjusting reserved part 31 is in a closed state, no air circulation exists in the recovery bin, so that the waste liquid in the reaction bin only enters the waste liquid pool 4 through the first flow channel and cannot enter the second flow channel. Similarly, after the extraction of nucleic acid is completed and the extracted nucleic acid needs to be recovered, the first air pressure regulation reservation part 31 is opened, the second air pressure regulation reservation part 41 is closed, the recovery bin enters air at the moment, the interior of the recovery bin is balanced with the atmospheric pressure, and the extracted nucleic acid enters the recovery bin through the second flow channel. The main purpose of the opening or closing of the second pneumatic adjustment allowance part 41 is to form a pressure difference inside the flow direction control system, and the liquid flows in different directions due to the pressure difference. The second air pressure adjustment allowance 41 may also be used in cooperation with a pressurizing member. The second air pressure regulating reserved part 41 may be a reserved hole on the waste liquid tank main body 4, or may be a branch pipe protruding from the waste liquid tank main body 4 as shown in fig. 1.

The top of the recovery bin main body 3 is provided with a first air pressure adjusting reserved part 31, and/or the top of the waste liquid pool main body 4 is provided with a second air pressure adjusting reserved part 41. The reason that the top was all located to two atmospheric pressure regulation preformed portion is that guarantee still can ventilate after there is liquid in recovery storehouse and waste liquid pond, and atmospheric pressure regulation preformed portion does not influence the liquid storage in retrieving storehouse and the waste liquid pond moreover, can not lead to the weeping.

The flow direction control system for the nucleic acid extraction device of the present invention can be used to constitute a nucleic acid extraction device.

Specifically, as shown in fig. 2 to 4, the nucleic acid extraction device is formed by matching a flow direction control system and a reagent bin.

In one embodiment, the nucleic acid extraction device is a disposable consumable with a flow direction control system.

In one embodiment, the method of using the flow direction control system for a nucleic acid isolation apparatus of the present invention is as follows:

sample adding: the sample of nucleic acid to be extracted is added to the reaction chamber through the reaction chamber air pressure adjusting section 21.

Cracking: the nucleic acid cracking reagent flows to the reaction bin, and is incubated with the sample in the reaction bin for mixed reaction, so that the sample is cracked by wall breaking.

Combining: the nucleic acid precipitation reagent and the magnetic bead suspension flow to the reaction chamber for reaction and combination for a certain time. The second air pressure regulation reserve part 41 is opened, the pressurizing member supplies air to the reaction chamber air pressure regulation part 21, and the generated waste liquid flows to the waste liquid pool through the first flow channel.

Washing: the rinsing liquid flows to the reaction chamber, the second air pressure regulation reserve part 41 is opened, the pressurizing member supplies air to the reaction chamber air pressure regulation part 21, and the generated waste liquid flows to the waste liquid tank through the first flow channel. The washing step is repeated.

Elution and recovery: the eluent 6 flows to the reaction chamber to elute the nucleic acid from the magnetic beads, the pressurizing member supplies air to the reaction chamber air pressure regulating portion 21, the first air pressure regulating reserve portion 31 is pierced, and the extracted nucleic acid flows into the recovery chamber through the second flow channel.

In conclusion, the invention is suitable for extracting and purifying nucleic acid from various samples of different types, has high integration level and easy use, effectively overcomes various defects in the prior art and has high industrial utilization value.

The foregoing embodiments are merely illustrative of the principles and utilities of the present invention and are not intended to limit the invention. Any person skilled in the art can modify or change the above-mentioned embodiments without departing from the spirit and scope of the present invention. Accordingly, it is intended that all equivalent modifications or changes which can be made by those skilled in the art without departing from the spirit and technical spirit of the present invention be covered by the claims of the present invention.

Claims

1. The utility model provides a nucleic acid is flow direction control system for extraction device, its characterized in that, flow direction control system for extraction device includes reaction storehouse main part (2), retrieves storehouse main part (3) and waste liquid pond main part (4), reaction storehouse main part (2) are through first runner main part (6) and waste liquid pond main part (4) intercommunication, reaction storehouse main part (2) are through second runner main part (7) and retrieve storehouse main part (3) intercommunication, be equipped with reaction storehouse atmospheric pressure regulating part (21) on reaction storehouse main part (2), retrieve storehouse main part (3) intercommunication first atmospheric pressure and adjust reservation portion (31), waste liquid pond main part (4) intercommunication second atmospheric pressure adjusts reservation portion (41).

2. The flow direction control system for a nucleic acid isolation apparatus according to claim 1, further comprising a support plate (5), wherein the reaction chamber body (2), the recovery chamber body (3), the waste liquid pool body (4), the first flow path body (6), and the second flow path body (7) are formed on the support plate (5).

3. The flow direction control system for a nucleic acid isolation apparatus according to claim 2, wherein a sealing film is provided on the support plate (5) and cooperates with the support plate (5) to form a reaction chamber, a recovery chamber, a waste liquid tank, a first flow channel and a second flow channel.

4. The system for controlling the flow direction of a nucleic acid isolation apparatus according to claim 3, wherein the support plate (5) is made of a flexible material, and the sealing film is flat.

5. The flow direction control system for the nucleic acid extraction device according to claim 1, wherein the second flow path main body (7) comprises a reaction bin output section (71), an upturned section (72), and a recovery bin input section (73) which are sequentially communicated.

6. The flow direction control system for a nucleic acid isolation apparatus according to claim 5, wherein the angle (α) between the reaction chamber output section (71) and the upwarp section (72) is an acute angle, and/or the angle (β) between the upwarp section (72) and the recovery chamber input section (73) is an acute angle.

7. The flow direction control system for a nucleic acid isolation apparatus according to claim 1, wherein an angle (γ) between the first flow channel main body (6) and the reaction chamber output section (71) is an obtuse angle.

8. The system for controlling a flow direction of a nucleic acid isolation apparatus according to claim 1, wherein the first flow channel and the waste liquid pool are located below the reaction chamber in a state where the system for controlling a flow direction of a nucleic acid isolation apparatus is used.

9. The flow direction control system for a nucleic acid isolation apparatus according to claim 1, wherein the flow direction control system for a nucleic acid isolation apparatus is further provided with a plurality of air bags or air pumps for supplying pneumatic pressure to the reaction chamber air pressure regulation section (21), the first air pressure regulation reserve section (31), or the second air pressure regulation reserve section (41).

10. The flow direction control system for a nucleic acid isolation apparatus according to claim 1, wherein the first air pressure regulation reserve part (31) is provided at the top of the recovery bin main body (3), and/or the second air pressure regulation reserve part (41) is provided at the top of the waste liquid pool main body (4).